Mutations in the sigma subunit of E. coli RNA polymerase which affect positive control of transcription.
The sigma subunits of bacterial RNA polymerases are required for the selective initiation of transcription. We have isolated and characterized mutations in rpoD, the gene which encodes the major form of sigma in E. coli, which affect the selectivity of transcription. These mutations increase the expression of araBAD up to 12-fold in the absence of CAP-cAMP. Expression of lac is unaffected, while expression of malT-activated operons is decreased. We determined the DNA sequence of 17 independently isolated mutations, and found that they consist of three different changes in a single CGC arginine codon at position 596 in the sigma polypeptide.


Relationship between feeding-induced ventilatory activity and bioaccumulation of dissolved and algal-bound cadmium in the Asiatic clam Corbicula fluminea.
The influence of feeding-induced ventilatory adaptation on cadmium accumulation patterns was analyzed in quiescent Asiatic clam Corbicula fluminea studied in steady-state conditions at 15 degrees C and 25 degrees C and in air-equilibrated water. Ventilatory activity, algae inflow rate, amounts of inspired dissolved and algae-bound Cd, and whole-body and tissue-specific metal concentrations were studied during a 15-d exposure period at a low dissolved cadmium concentration of 2 microg/L and various algal concentrations of Scenedesmus subspicatus. Great care was taken not to externally stimulate animals. Four main results were obtained as regard to regulation of ventilatory activity and metal accumulation rate. First, at 25 degrees C, feeding purposes strongly stimulated ventilatory activity, while at 15 degrees C, it was independent of it except at 1 to 2 x 10(5) algae/ml. Second, Cd accumulation rate increased at higher temperature. Third, accumulation rate exhibited a positive correlation with the ventilatory flow rate. Fourth, no correlation was observed with the quantity of ventilated algae-bound Cd but, rather, with the quantity of ventilated dissolved Cd. It is concluded that the physiology of the animals can be fundamental in affecting metal accumulation process.


BET bromodomain inhibition suppresses adipogenesis in mice.
We recently reported that inhibition of BET bromodomain suppresses adipogenesis in vitro. In the present study we aimed to address whether BET bromodomain inhibition can suppress adipogenesis in vivo. Brd4fl/fl mice were crossed with B6.Cg-Tg(Fabp4-cre)1Rev/J mice to generate Brd4fl/+/Fabp4-cre mice. We used high fat diet (HFD, 45% fat) mice treated with vehicle (DMSO) or JQ1 (intraperitoneal, IP injection, 50 mg/kg/day), respectively, for 6 weeks. Body weight was measured once a week. Dual-energy X-ray absorptiometry was determined and brown adipose tissue was harvested at the end of the experiment. Partial deletion of Brd4 leads to the lower body weight. JQ1 treatment further confirmed that BET bromodomain inhibition suppresses body weight gain and decreases white adipose depots compared with the control mice. In addition, JQ1 treatment reduces the size of brown adipose tissue and impairs its thermogenesis. BET bromodomain inhibition suppresses adipogenesis in the mice.


Herpes simplex virus replication in pheochromocytoma cell line that responds to nerve growth factor.
Cultured cells of neural origin (PC-12, pheochromocytoma cell line) respond to nerve growth factor (NGF) by extending neurites. These cells whether treated with NGF or not can be infected with herpes simplex virus and produce progeny virus. Viral antigens are detected on the cell surface and fusion of cells to form polykaryocytes takes place. Nucleocapsids are found within the cell nucleus and enveloped virus in a present both in the cytoplasm and extracellular space. Virus was not observed within the neurites but laterations in the neurite microtubular structure occurred after infection.


Construction of a risk model based on N6 -methyladenosinerelated lncRNAs for predicting the prognosis of breast cancer.
N6-methyladenosine modification and lncRNAs are closely related to the prognosis and immunotherapy response of breast cancer patients. LncRNAs related to m6 A-associated genes were predicted based on coexpression analysis of the TCGA database. We established a novel 7-m6 A-associated lncRNA signature for predicting patient prognosis and validated it. The model was significantly correlated with survival time and survival status and was an independent predictor of overall survival (OS). Except for the M1 disease group, the model had good predictive value for OS in different subgroups. We constructed a prognostic model based on 7 m6 A-associated lncRNAs in breast cancer. This model could serve as an independent prognostic factor with tremendous predictive ability for breast cancer patients.


Anti-oxidative feedback and biomarkers in the intertidal seagrass Zostera japonica induced by exposure to copper, lead and cadmium.
To investigate the potential influences of anthropogenic pollutants, we evaluated the responses of the intertidal seagrass Zostera japonica to three heavy metals: copper (Cu), lead (Pb), and cadmium (Cd). Z. japonica was exposed to various concentrations of Cu, Pb, and Cd (0, 0.5, 5, 50μM) over seven days. The effects were then analyzed using the antioxidant enzyme catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), glutathione peroxidase (GPX), and lipid peroxidation measured using malondialdehyde (MDA) as proxy. Metal accumulation in the above-ground tissues and phenotypic changes were also investigated. Our results revealed that heavy metal concentration increased in seagrass exposed to high levels of metals. Z. japonica has great potential for metal accumulation and a suitable candidate for the decontamination of moderately Cu contaminated bodies of water and can also potentially enhanced efforts of environmental decontamination, either through phytoextraction abilities or by functioning as an indicator for monitoring programs that use SOD, CAT, GPX, POD and MDA as biomarkers.


Impact of Attending Physicians' Comments on Residents' Workloads in the Emergency Department: Results from Two J(^o^)PAN Randomized Controlled Trials.
To examine whether peppy comments from attending physicians increased the workload of residents working in the emergency department (ED). We conducted two parallel-group, assessor-blinded, randomized trials at the ED in a tertiary care hospital in western Japan. Twenty-five residents who examined either ambulatory (J(^o^)PAN-1 Trial) or transferred patients (J(^o^)PAN-2 Trial) in the ED on weekdays. Participants were randomly assigned to groups that either received a peppy message such as "Hope you have a quiet day!" (intervention group) or did not (control group) from the attending physicians. Both trials were conducted from June 2014 through March 2015. For each trial, residents rated the number of patients examined during and the busyness and difficulty of their shifts on a 5-point Likert scale. A total of 169 randomizations (intervention group, 81; control group, 88) were performed for the J(^o^)PAN-1 Trial, and 178 (intervention group, 85; control group, 93) for the J(^o^)PAN-2 Trial. In the J(^o^)PAN-1 trial, no differences were observed in the number of ambulatory patients examined during their shifts (5.5 and 5.7, respectively, p = 0.48), the busyness of their shifts (2.8 vs 2.8; p = 0.58), or the difficulty of their shifts (3.1 vs 3.1, p = 0.94). However, in the J(^o^)PAN-2 trial, although busyness (2.8 vs 2.7; p = 0.40) and difficulty (3.1 vs 3.2; p = 0.75) were similar between groups, the intervention group examined more transferred patients than the control group (4.4 vs 3.9; p = 0.01). Peppy comments from attending physicians had a minimal jinxing effect on the workload of residents working in the ED. University Hospital Medical Information Network Clinical Trials Registry (UMIN-CTR), UMIN000017193 and UMIN000017194.


Neuroimaging of reward mechanisms in Gambling disorder: an integrative review.
Gambling disorder (GD) was reclassified as a behavioral addiction in the DSM-5 and shares clinical and behavioral features with substance use disorders (SUDs). Neuroimaging studies of GD hold promise in isolating core features of the addiction syndrome, avoiding confounding effects of drug neurotoxicity. At the same time, a neurobiologically-grounded theory of how behaviors like gambling can become addictive remains lacking, posing a significant hurdle for ongoing decisions in addiction nosology. This article integrates research on reward-related brain activity (functional MRI) and neurotransmitter function (PET) in GD, alongside the consideration of structural MRI data as to whether these signals more likely reflect pre-existing vulnerability or neuroadaptive change. Where possible, we point to qualitative similarities and differences with established markers for SUDs. Structural MRI studies indicate modest changes in regional gray matter volume and diffuse reductions in white matter integrity in GD, contrasting with clear structural deterioration in SUDs. Functional MRI studies consistently identify dysregulation in reward-related circuitry (primarily ventral striatum and medial prefrontal cortex), but evidence is mixed as to the direction of these effects. The need for further parsing of reward sub-processes is emphasized, including anticipation vs outcome, gains vs. losses, and disorder-relevant cues vs natural rewards. Neurotransmitter PET studies indicate amplified dopamine (DA) release in GD, in the context of minimal differences in baseline DA D2 receptor binding, highlighting a distinct profile from SUDs. Preliminary work has investigated further contributions of opioids, GABA and serotonin. Neuroimaging data increasingly highlight divergent profiles in GD vs. SUDs. The ability of gambling to perpetually activate DA (via maximal uncertainty) may contribute to neuroimaging similarities between GD and SUDs, whereas the supra-physiological DA effects of drugs may partly explain differences in the neuroimaging profile of the two syndromes. Coupled with consistent observations of correlations with gambling severity and related clinical variables within GD samples, the overall pattern of effects is interpreted as a likely combination of shared vulnerability markers across GD and SUDs, but with further experience-dependent neuroadaptive processes in GD.


Mutations in the NEB gene cause fetal akinesia/arthrogryposis multiplex congenita.
We studied a series of patients with fetal akinesia deformation sequence (FADS)/arthrogryposis multiplex congenita (AMC), with nemaline bodies on muscle specimens, which revealed mutations in the NEB gene. We pathologically assessed seven cases from three families, who presented with AMC/FADS. Targeted genetic analysis for Ashkenazi Jewish mutation (in relevant patients) was followed by next-generation sequencing and multiplex ligation-dependent probe amplification. All cases were detected on prenatal ultrasound. Characteristic nemaline bodies on muscle specimens were demonstrated in at least one case in each of the nuclear families. In the Ashkenazi Jewish family, the known founder mutation was compounded by one recurrent novel splice site. The other two families were of Chinese and Korean origins, and only one pathogenic heterozygous mutation was detected in each. Nemaline myopathy due to NEB mutation(s) leads to FADS/AMC. Currently, mutated NEB is under-recognized as a cause for AMC/FADS. Our study attempts to raise recognition of this gene as a cause, suggesting the NEB gene should be included in genetic panels used for FADS/AMC cases and be fully covered when EXOME sequencing is utilized. A heterozygous mutation may suggest either compounding undetected one or digenic interaction that requires further genetic analyses. © 2016 John Wiley & Sons, Ltd.


Urinalysis in patients with neuromyelitis optica spectrum disorder.
Increasing evidence has demonstrated that aquaporin-4 (AQP4) immunoglobulin G causes damage to the kidney in neuromyelitis optica spectrum disorder (NMOSD). However, changes in urinalysis in NMOSD have not been investigated thus far. Our objective was to evaluate the changes in urinalysis in NMOSD patients. Case data were collected from 44 patients with AQP4 antibody-positive NMOSD, 53 patients with multiple sclerosis (MS) and 79 age- and sex-matched healthy controls. Analyses of early morning urine and 24-h urine samples comparing NMOSD with MS patients were conducted. In the acute phase, urine pH levels (P < 0.001) and urine specific gravity levels (P < 0.001) from NMOSD patients were significantly higher and lower, respectively, than for MS patients. 24-h urine sodium and 24-h urine volume from NMOSD patients were significantly higher than for MS patients (both P = 0.001). A 24-h urine volume higher than 2500 ml (odds ratio 11.7, 95% confidence interval 1.863-73.066) and a 24-h urine sodium higher than 200 mmol (odds ratio 16.0, 95% confidence interval 2.122-120.648) are more likely to occur in NMOSD patients in the acute phase than in MS patients. The urinalysis results were significantly different between NMOSD patients and MS patients. The pathophysiological changes in AQP4 antibody-positive NMOSD patients were not limited to the central nervous system.


Retinal morphology and visual pigment levels in 6- and 12-month-old rhesus monkeys fed a taurine-free human infant formula.
Rhesus monkey infants were raised from birth until 6 or 12 months of age on a taurine-free soy protein-based human infant formula or on the same formula supplemented with taurine. An additional group received taurine-free formula until 6 months and then the supplemented diet from 6 until 12 months. The densities of rod and cone visual pigments were measured by fundus reflectometry at 6 and 12 months, and retinal morphology was then examined by light and electron microscopy. The densities of rhodopsin, measured in the near periphery after a white bleach, and of cone pigment, measured in the macula after a red bleach, were significantly reduced in the taurine-deprived monkeys at 6 months but not at 12 months. The retinas of 6-month-old taurine-deprived infants showed degenerative morphological changes in photoreceptors, particularly in cones in the foveal region, which were somewhat less severe than those seen in a previous study at 3 months of age. The prevalence and degree of these abnormalities continued to decrease with age in taurine-deprived animals, but changes persisted in some animals at 12 months. Recovery was more complete in monkeys reversed to the supplemented diet from 6 to 12 months. Thus, monkey infants are dependent on dietary taurine to maintain normal retinal structure until at least 6 months of age; the effects of taurine deprivation regress spontaneously but incompletely by 12 months.


Enhanced analgesic effect of morphine-nimodipine combination after intraspinal administration as compared to systemic administration in mice.
Calcium plays an important role in the pathophysiology of pain. A number of studies have investigated the effect of L-type calcium channel blockers on the analgesic response of morphine. However, the results are conflicting. In the present study, the antinociceptive effect of morphine (2.5 microg) and nimodipine (1 microg) co-administered intraspinally in mice was observed using the tail flick test. It was compared to the analgesic effect of these drugs (morphine - 250 microg subcutaneously; nimodipine - 100 microg intraperitoneally) after systemic administration. Nimodipine is highly lipophilic and readily crosses the blood brain barrier. Addition of nimodipine to morphine potentiated the analgesic response of the latter when administered through the intraspinal route but not when administered through systemic route. It may be due to direct inhibitory effect of morphine and nimodipine on neurons of superficial laminae of the spinal cord after binding to mu -opioid receptors and L-type calcium channels respectively.


Effects of ceftriaxone induced intestinal dysbacteriosis on lymphocytes in different tissues in mice.
The close relationship between intestinal microflora and immune system has been confirmed, stimulus from intestinal flora plays an important role in the development of the immune system and its dynamic balance. Current research is still inadequate to determine how local bacteria in gut influence the whole body. In this study, influence of ceftriaxone sodium induced intestinal dysbacteriosis on local and overall immune function was investigated. We found that the beneficial bacteria decreased significantly compared with control after oral administration of ceftriaxone; Moreover, the proportion of T cells are higher and B cells are lower in the dysbacteriosis mice, activation and proliferation of T and B cells was decreased significantly in gut-associated lymphoid tissues (GALTs), such as Peyer's patches (PPs) and mesenteric lymph nodes(MLNs)with ceftriaxone treatment; The secreted sIgA in intestinal was reduced in dysbacteriosis mice than that of control as well. The similar results above are also shown on the spleen. In addition, the delayed type hypersensitivity (DTH) reaction decreased in dysbacteriosis mice. The present data suggested that intestinal microflora had impact on immune system by influencing the proportion and function of lymphocytes in PPS-MLN-spleen.


Is diabetic striatopathy the culprit of seizures in a patient with ketotic hyperglycemia-induced hemichorea-hemiballismus?
Diabetic striatopathy is a rare neurological manifestation of nonketotic hyperglycemia that presents with contralateral hemichorea-hemiballismus. Presentation with concurrent seizures is rarely reported. We report a case of diabetic striatopathy presenting with focal and generalized tonic-clonic seizures (GTCS) with right hemichorea-hemiballismus induced by a ketotic hyperglycemic state. Head MRI showed high T1-weighted signal intensity in the left lentiform nucleus with no significant diffusion restriction or postcontrast enhancement. The patient's condition gradually improved, with seizure control on AEDs. Hemichorea-hemiballismus significantly improved with adequate blood sugar control and resolved with low-dose haloperidol. Diabetic striatopathy presenting with hemichorea-hemiballismus and concurrent GTCS has been reported previously in two cases; however, it has never been reported in ketotic hyperglycemia. To the best of our knowledge, we herein report the first case report of focal and generalized seizures in a ketotic hyperglycemic state and mesial temporal sclerosis.


Treatment of chronic type B aortic dissection by endovascular grafting in a previously CABG patient. Case report.
High mortality rates resulting from the surgical treatment of Type B aortic dissections have played an important role in seeking newer alternative therapy modalities for the treatment of the pathology. Especially since the early 1990s, endovascular grafting has become a popular option in the treatment of aortic diseases and with the increasing experience and advances in technology, it has, in recent years, become an alternative approach for the treatment of aortic dissections. In this report, we present the treatment of a 56-year-old male patient with a history of a coronary artery bypass grafting who presented with chronic type B aortic dissection. Endovascular stent graft implantation was performed with a hybrid therapy type of preliminary right subclavian artery to left subclavian artery bypass, followed by endovascular stent graft implantation to the dilated aneurismal segment of the descending aorta, for the treatment of chronic type B aortic dissection. The procedure and the postoperative course were uneventful; moreover, they were very comfortable for the patient undergoing an investigation of the descending aorta, when compared with the conventional surgical treatment modalities.


Versatility and specialization in labrid fishes: ecomorphological implications.
The term "specialized" has been used to describe species that possess unique functional attributes and/or a narrow, stereotyped range of attributes, but there are few comparative functional analyses of specialists and generalists. If species with functional morphological specializations are capable of functioning over a broad range, the link between morphology and ecology may be relaxed under certain environmental conditions. In this study, high-speed films of jaw movements during prey capture were compared statistically for three coexisting coral reef fish species in the family Labridae, one trophic specialist and two trophic generalists. The trophic specialist possessed a unique functional feature related to the movement of the hyoid in the floor of the mouth, while the trophic generalists were not observed to possess any functional specializations. All three species showed functional versatility in that they were able to adjust their prey capture mechanism in response to the evasive potential of the prey. The functional versatility of trophic specialists has implications for ecomorphological studies, since species characterized as possessing unique functional or morphological features may demonstrate marked flexibility in ecological variables such as diet or foraging behavior, decreasing the likelihood of identifying correlations between morphology and ecology.


A comparison of six analytical disease mapping techniques as applied to West Nile Virus in the coterminous United States.
West Nile Virus has quickly become a serious problem in the United States (US). Its extremely rapid diffusion throughout the country argues for a better understanding of its geographic dimensions. Both 2003 and 2004 percentages of deaths by numbers of reported human cases, for the 48 coterminous US states, are analyzed with a range of spatial statistical models, seeking to furnish a fuller appreciation of the variety of models available to researchers interested in analytical disease mapping. Comparative results indicate that no single spatial statistical model specification furnishes a preferred description of these data, although normal approximations appear to furnish some questionable implications. Findings also suggest several possible future research topics.


Strategies to obtain multiple recombinant modified vaccinia Ankara vectors. Applications to influenza vaccines.
As a vaccination vector, MVA has been widely investigated both in animal models and humans. The construction of recombinant MVA (rMVA) relies on homologous recombination between an acceptor virus and a donor plasmid in infected/transfected permissive cells. Our construction strategy "Red-to-Green gene swapping" - based on the exchange of two fluorescent markers within the flanking regions of MVA deletion ΔIII, coupled to fluorescence activated cell sorting - is here extended to a second insertion site, within the flanking regions of MVA deletion ΔVI. Exploiting this strategy, both double and triple rMVA were constructed, expressing as transgenes the influenza A proteins HA, NP, M1, and PB1. Upon validation of the harbored transgenes co-expression, double and triple recombinants rMVA(ΔIII)-NP-P2A-M1 and rMVA(ΔIII)-NP-P2A-M1-(ΔVI)-PB1 were assayed for in vivo immunogenicity and protection against lethal challenge. In vivo responses were identical to those obtained with the reported combinations of single recombinants, supporting the feasibility and reliability of the present improvement and the extension of Red-to-Green gene swapping to insertion sites other than ΔIII.


Chromosomal breaks at the origin of small tandem DNA duplications.
Small tandem DNA duplications in the range of 15 to 300 base-pairs play an important role in the aetiology of human disease and contribute to genome diversity. Here, we discuss different proposed mechanisms for their occurrence and argue that this type of structural variation mainly results from mutagenic repair of chromosomal breaks. This hypothesis is supported by both bioinformatical analysis of insertions occurring in the genome of different species and disease alleles, as well as by CRISPR/Cas9-based experimental data from different model systems. Recent work points to fill-in synthesis at double-stranded DNA breaks with complementary sequences, regulated by end-joining mechanisms, to account for small tandem duplications. We will review the prevalence of small tandem duplications in the population, and we will speculate on the potential sources of DNA damage that could give rise to this mutational signature. With the development of novel algorithms to analyse sequencing data, small tandem duplications are now more frequently detected in the human genome and identified as oncogenic gain-of-function mutations. Understanding their origin could lead to optimized treatment regimens to prevent therapy-induced activation of oncogenes and might expose novel vulnerabilities in cancer.


Dissemination of antibiotic resistance genes from landfill leachate to groundwater.
Landfill leachate, a highly concentrated organic wastewater containing diverse microorganisms and various heavy metals, has become an important reservoir of antibiotic resistance genes (ARGs). In this study, a total of 203 unique ARGs and 10 mobile genetic elements (MGEs) were identified from collected landfill leachate and groundwater. The number and abundance (normalized and absolute) of antibiotic resistome in effluent of leachate treatment plants decreased significantly compared to influent. The abundance of ARGs in groundwater increased as the distance from the leachate basin decreased. Fast expectation-maximization microbial source tracking (FEAST) showed that up to 96 % of ARGs in groundwater (GW3) may originate from nearby leachate, suggesting that ARGs in leachate can penetrate and spread into the groundwater environment. A significant correlation between ARGs and bacterial communities was identified. Together with network analysis showing the 12 bacterial taxa co-occurring with seven classes of antibiotic-associated ARGs, our results revealed the diverse potential microbial hosts of ARGs in water samples around the landfill sites. Heavy metals, bacterial community and MGEs were the driving factors shaping the ARGs patterns in the water samples, with their interactions explaining 57 % of ARGs variations. Our results provide an understanding of the distribution and dissemination of ARGs from landfill leachate to the nearby groundwater and suggest a comprehensive impact assessment of ARGs in aquatic environments of landfills.


Effects of c-Jun and a negative dominant mutation of c-Jun on differentiation and gene expression in lens epithelial cells.
We have used a retroviral vector (RCAS) to overexpress wild-type chicken c-Jun or a deletion mutant of chicken c-Jun (Jun delta 7) lacking the DNA binding region to investigate the possible role of c-Jun in lens epithelial cell proliferation and differentiation. Both constructs were efficiently expressed in primary cultures of embryonic chicken lens epithelial cells. Overexpression of c-Jun increased the rate of cell proliferation and greatly delayed the appearance of "lentoid bodies," structures which contain differentiated cells expressing fiber cell markers. Excess c-Jun expression also significantly decreased the level of beta A3/A1-crystallin mRNA, without affecting alpha A-crystallin mRNA. In contrast, the mutated protein, Jun delta 7, had no effect on proliferation or differentiation but markedly increased the level of alpha A-crystallin mRNA in proliferating cell cultures. These results suggest that c-Jun or Jun-related proteins may be negative regulators of alpha A- and beta A3/A1-crystallin genes in proliferating lens cells.


Acupuncture anaesthesia for squint surgery.
An account of an operation for squint using acupuncture analgesia is given. This is the first ophthalmic operation carried out in Australia using this form of analgesia; and is probably the first surgical operation of any type carried out in Australia using acupuncture technique. The indications and possibilities of this method are discussed.


Stability, speciation and spectral properties of NpO2+ complexes with pyridine monocarboxylates in aqueous solution.
Neptunyl ion as NpO2+ is the least reacting and most mobile radioactive species among all the actinides. The picolinic acid used for decontamination is co-disposed along with the radioactive waste. Thus, in long term storage of HLW, there is high possibility of interaction of actinides and long lived fission products with the picolinate and can cause migration. The complexation of NpO2+ with the three structural isomers of pyridine monocarboxylates provides an insight to explore the role of hetero atom (nitrogen) with respect to key binding moiety (carboxylate). In the present study, the log β values, speciation and spectral properties of NpO2+ complexes with pyridine monocarboxylates viz. picolinate, nicotinate and isonicotinate, have been studied at 298K in 0.1M NaClO4 medium using spectrophotometry. The complexation reactions involving protonated ligands are always accompanied by protonation/deprotonation process; thus, the protonation constants of all the three pyridine monocarboxylates under same conditions were also determined by potentiometry. The spectrophotometric data analysis for complexation of NpO2+ with pyridine monocarboxylates indicated the presence of ML and ML2 complexes with log β values of 2.96±0.04, 5.67±0.08 for picolinate, 1.34±0.09, 1.65±0.12 for nicotinate and 1.52±0.04, 2.39±0.06 for isonicotinate. The higher values of log β for picolinate were attributed to chelation while in other two isomers, the binding is through carboxylate group only. Density Functional Theory (DFT) calculations were carried out to get optimized geometries and electrostatic charges on various atoms of the complexes and free pyridine monocarboxylates to support the experimental data. The higher stability of NpO2+ nicotinate and isonicotinate complexes compared to simple carboxylates and the difference in log β between the two is due to the charge polarization from unbound nitrogen to the bound carboxylate oxygen atoms.


Influence of microwave temperature and power on the biomethanation of food waste under mesophilic anaerobic conditions.
Food waste is an attractive feedstock for Anaerobic Digestion due to its high biodegradability and moisture content. Nevertheless, due to its complex structure and composition, methane yield is typically compromised with 50-60% of the theoretical maximum obtained. The well-known limitation of the hydrolysis step can be circumvented by adopting feedstock pre-treatments, such as microwave irradiation. It improves solubilization of various FW components making them more readily available for the microorganisms and reducing AD process duration. In this work different heating rates (7.8, 3.9 and 1.9 °C/min) and temperatures (85, 115, 145, 175 °C) were applied when pre-treating food waste as a substrate for AD. Increase in the solubilization of organic matter in the form of Soluble Chemical Oxygen Demand was the most significative change in FW characteristics after pre-treatment, with final temperature of 175 °C and heating rate of 3.9 °C showing a 73.19% increment. Nevertheless, process performance of AD of MW FW was optimum at 85 °C 7.8 ramp, showing no intermediate products accumulation, up to 77% more methane produced in the first week of digestion compared to the other conditions tested and reduction of 96.36% on the lag phase duration, compared to the control. On the other hand, samples treated at 175 °C, regardless of heating rate, consistently showed poor process performance, with low methane yield, possibly due to the formation of hard-to-digest compounds. This work underlines the importance of adjusting microwave temperature and power when pre-treating FW for biomethane production so the process is optimized.


A piRNA to remember.
In this issue of Cell, Rajasethupathy et al. report a surprising role for piRNAs, previously thought to act mainly in the animal germline to silence transposons, in transcriptional regulation of plasticity-related genes in the central nervous system of the sea slug Aplysia californica. The findings expand the functions of small RNAs and have important implications for our understanding of how transient signals can give rise to long-term memories.


Advanced throughput analytical strategies for the comprehensive HRMS screening of organic micropollutants in eggs of different bird species.
Raptors are ideal indicators for biomonitoring studies using wildlife in order to assess the environmental pollution in the terrestrial ecosystem, since they are placed in the highest trophic position in the food webs and their life expectancy is relatively long. In this study, 26 eggs of 4 bird species (Peregrine falcon, Eurasian curlew, Little owl and Eagle owl) collected in Germany, were investigated for the presence of persistent organic pollutants (POPs) and thousands of contaminants of emerging concern (CECs). Generic sample preparation protocols were followed for the extraction of the analytes and the purification of the extracts, and the samples were analyzed both by liquid (LC) and gas chromatography (GC) coupled to high resolution mass spectrometry (HRMS), for capturing a wide range of organic micropollutants with different physicochemical properties. State-of-the-art screening methodologies were applied in the acquired HRMS data, including wide-scope target analysis of 2448 known pollutants and suspect screening of over 65,000 environmentally relevant compounds. Overall, 58 pollutants from different chemical classes, such as plant protection products, per- and polyfluoroalkyl substances and medicinal products, as well as their transformation products, were determined through target analysis. Most of the detected compounds were lipophilic (logP>2), although the presence of (semi)polar contaminants should not be overlooked, underlying the need for holistic analytical approaches in environmental monitoring studies. p,p'-DDE, PCB 153 and PCB138, PFOS and methylparaben were the most frequently detected compounds. 50 additional substances were identified and semi-quantified through suspect screening workflows, including mainly compounds of industrial use with high production volume.


Changes in cAMP phosphodiesterase activity and cAMP concentration during mouse preimplantation development.
Cyclic nucleotide phosphodiesterase (PDE) activity and cAMP amounts were measured in mouse preimplantation embryos at the 1-cell, 2-cell, 8-cell/morula, and mid-blastocyst stages. PDE activity remained constant between the 1-cell and 2-cell stages. It decreased by the 8-cell stage and continued to decrease by the mid blastocyst stage to about 14% of the 1- and 2-cell values. By contrast, cAMP amounts remained essentially constant at 0.05 fmole/embryo (0.3 microM) from the 1-cell to the blastocyst stage and increased to 0.175 fmole in the fully expanded blastocyst that was close to hatching. Measurements of embryo volume indicated that intracellular volume remained essentially constant up to the blastocyst stage. The morphological changes in cell shape that accompany differentiation of the trophectoderm and that are coupled with blastocoel expansion decreased the intracellular volume. This decrease resulted in an increase in the cAMP concentration to about 0.4 microM by the mid-blastocyst stage. Previous studies indicate that either cAMP or TGF-alpha/EGF can stimulate the rate of blastocoel expansion. Although TGF-alpha/EGF can elevate cAMP levels in other cell types, TGF-alpha, at a concentration that maximally stimulates the rate of blastocoel expansion, did not elevate cAMP in blastocysts. Thus, it was unlikely that elevation of cAMP is the mechanism by which TGF-alpha stimulates the rate of blastocoel expansion.


Relaxation of corpus cavernosum and raised intracavernous pressure by berberine in rabbit.
1. In the present study, we have investigated the effect of berberine in rabbit isolated corpus cavernosum and measured the intracavernous pressure (ICP) change after intracavernosal injection of berberine in rabbit. 2. Berberine alone suppressed the basal tone and induced a concentration (0.1-100 microM)-dependent relaxation in phenylephrine (PE)-precontracted corpus cavernosum. 3. Tetrodotoxin (0.1 and 1 microM) treatment had no significant effect on the berberine-induced relaxation. Phentolamine (1 and 10 microM), propranolol (1 and 3 microM) and atropine (1 and 3 microM) were also without effect. These results suggest that berberine might cause relaxation of the cavernosal strip by direct action on the corpus cavernosum, not by a neuronal effect. Furthermore, muscarinic- and beta-adrenoceptors were not involved. 4. Berberine-induced relaxations were significantly reduced by endothelium removal and by exposure to L-NG-nitro arginine methyl ester (0.1 and 0.3 mM), but not indomethacin (30 microM). 5. In endothelium-deprived corpus cavernosal tissues, berberine-induced relaxations were significantly reduced in high K+ medium (KCl = 60 mM), by charybdotoxin (ChTX) and 4-aminopyridine (4-AP) but not by glibenclamide and apamin. 6. After intracavernous injection of berberine (1, 2, 3 and 5 mg kg(-1)), the ICP rose from 12.7+/-3.6 to 13.2+/-5.4, 25.3+/-6.1, 46.5+/-8.2, and 63.4+/-10.2 mmHg, respectively. The duration of tumescence ranged from 11.5 - 43.7 min. 7. The results show that berberine possesses a relaxant effect on rabbit corpus cavernosal tissues which is attributable to both endothelium-dependent and-independent properties. While the former component is apparently due to the release of NO from sinusoidal endothelium, the endothelium-independent mechanism involved in berberine relaxation is probably linked to ChTX- and 4-AP-sensitive K+ channel activation in the cavernosal vasculature.


Alcoholism and abstinence among relatives of American Indian alcoholics.
The incidence of alcoholism among relatives of Indian alcoholics was significantly greater than it was among relatives of non-Indian alcoholics.


Immunologic studies in cow's milk-induced pulmonary hemosiderosis.
Antibodies to cow's milk proteins (CMP) were studied by radioallergosorbent tests to determine the quantity and immunoglobulin class of these antibodies in nine patients with precipitins to cow's milk. Three of these patients had pulmonary hemosiderosis (PH); in one other patient pulmonary hemosiderosis was suspected, but not proven. Three patients had nonhemosiderosis chronic lung disease and two had other diseases. Age-matched control subjects were also studied. The quantity and immunoglobulin class distribution of antibodies to CMP were similar in all patients; the quantity of anti-CMP antibody was significantly greater in patients than in control subjects (P less than 0.001). In addition, complement-fixing antibody to CMP was not detected in either patients' or controls' sera. Lymphocyte responses to CMP were studied in three patients with PH, three individuals with other manifestations of milk hypersensitivity (positive controls), and nine negative control subjects. Patients and positive control subjects responded to CMP with greater tritiated thymidine incorporation than the negative control subjects (P less than 0.05). However, no significant difference in response to CMP was observed between the PH patients and the positive control subjects. Antibodies to human lung tissue were studied in the sera of all patients with PH and two negative control subjects. Antibodies to human lung and rat lung were detected in one patient with PH.


Resolution of elevated postvoid residual volumes after correction of severe pelvic organ prolapse.
We hypothesized that elevated postvoid residual volume (PVRV) would resolve postoperatively in women with severe pelvic organ prolapse (POP) and investigated risk factors that might hamper its resolution. Retrospectively, we enrolled 57 patients undergoing surgery for severe POP. All patients had preoperative PVRV > or = 100 ml documented by ultrasound and catheterization. Presurgical voiding difficulty, urodynamic and cystoscopic data, and surgical outcome were analyzed. The incidence of postsurgical elevated PVRV was 15.8% and 3.5% was symptomatic. Patients with postsurgical elevated PVRV had larger preoperative residual volume than those with normal postsurgical PVRV (P = 0.037). By multivariable analysis, concomitant anti-incontinence surgery was the single independent predictor of postsurgical elevated PVRV (odds ratio = 5.38, P = 0.031). A majority of patients with severe POP had their elevated PVRV resolved postoperatively. Although concomitant anti-incontinence surgery increased the risk of developing elevated PVRV after repair, most remained asymptomatic.


Responses of antibiotics, antibiotic resistance genes, and mobile genetic elements in sewage sludge to thermal hydrolysis pre-treatment and various anaerobic digestion conditions.
Sewage sludge from wastewater treatment plants (WWTPs) harbours large amounts of antibiotics, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs), the variation and fate of these emerging pollutants during sludge treatment processes must be thoroughly studied to reduce their potential risks to human health. In this study, 7 pilot-scale CSTR anaerobic digesters were established with the same seed sludge and fed with the same thermal hydrolysis pre-treated sewage sludge, while operating under different conditions. High-throughput quantitative PCR, UPLC-MS/MS and Illumina Hiseq-sequencing were used to systematically evaluate the responses of antibiotics, ARGs, and MGEs in sewage sludge to thermal hydrolysis pre-treatment and various anaerobic digestion (AD) conditions. The results showed that thermal hydrolysis effectively reduced the abundance (>94%) of almost all subtypes of ARGs and MGEs, and it was a powerful technology for reducing tetracyclines, macrolides, and lincosamides. Besides, the abundance of ARGs and MGEs in thermophilic digesters was lower than that in mesophilic digesters, suggesting that thermophilic digesters could be used to avoid the ARGs rebounding. In addition, the thermophilic system further reduced the concentrations of quinolones. For the digesters operated under the mesophilic conditions, a longer hydraulic retention time (HRT) facilitated the removal of antibiotics, ARGs, and MGEs. Furthermore, the microbial community and MGEs had important effects on the persistence and proliferation of ARGs in AD process. The findings of this study provide effective clues for controlling the spread of antibiotic resistance and suggest the optimal operating conditions of digesters.


[Field 8. Safety practices in paediatric intensive care medicine. French-speaking Society of Intensive Care. French Society of Anesthesia and Resuscitation].
A paediatric intensive care unit is a complex environment with many hazards for potential medical errors. Due to the high acuity of illness and the complexity of medical care with multiple disciplines and individuals involved, medical errors occur due to breakdowns in communication and teamwork. Medication errors are the most frequent errors in paediatrics. This chapter proposes some strategies to prevent medical errors in paediatric intensive care units.


3D convolutional neural networks-based multiclass classification of Alzheimer's and Parkinson's diseases using PET and SPECT neuroimaging modalities.
Alzheimer's disease (AD) is a neurodegenerative brain pathology formed due to piling up of amyloid proteins, development of plaques and disappearance of neurons. Another common subtype of dementia like AD, Parkinson's disease (PD) is determined by the disappearance of dopaminergic neurons in the region known as substantia nigra pars compacta located in the midbrain. Both AD and PD target aged population worldwide forming a major chunk of healthcare costs. Hence, there is a need for methods that help in the early diagnosis of these diseases. PD subjects especially those who have confirmed postmortem plaque are a strong candidate for a second AD diagnosis. Modalities such as positron emission tomography (PET) and single photon emission computed tomography (SPECT) can be combined with deep learning methods to diagnose these two diseases for the benefit of clinicians. In this work, we deployed a 3D Convolutional Neural Network (CNN) to extract features for multiclass classification of both AD and PD in the frequency and spatial domains using PET and SPECT neuroimaging modalities to differentiate between AD, PD and Normal Control (NC) classes. Discrete Cosine Transform has been deployed as a frequency domain learning method along with random weak Gaussian blurring and random zooming in/out augmentation methods in both frequency and spatial domains. To select the hyperparameters of the 3D-CNN model, we deployed both 5- and 10-fold cross-validation (CV) approaches. The best performing model was found to be AD/NC(SPECT)/PD classification with random weak Gaussian blurred augmentation in the spatial domain using fivefold CV approach while the worst performing model happens to be AD/NC(PET)/PD classification without augmentation in the frequency domain using tenfold CV approach. We also found that spatial domain methods tend to perform better than their frequency domain counterparts. The proposed model provides a good performance in discriminating AD and PD subjects due to minimal correlation between these two dementia types on the clinicopathological continuum between AD and PD subjects from a neuroimaging perspective.


Presentation of a radicular cyst associated with a primary molar.
A radicular cyst in a 9-year old girl is reported. A 9-year-old child was referred from her General Dental Practitioner because of the need for obtaining expert care, assessment and treatment with regards to a radiolucency associated with her lower right second primary molar (85). This included surgical intervention for the removal of the affected tooth, enucleation of the cyst and primary closure of the surgical area. This was satisfactory and 12 months post-operative the patient reported no further problems or symptoms. It could be seen that the area previously occupied by the cyst had undergone bony infill.


Sirolimus and tacrolimus coefficient of variation is associated with rejection, donor-specific antibodies, and nonadherence.
Immunosuppression medication nonadherence has been associated with donor-specific antibodies and treatment-refractory rejection. Drug-level monitoring is a practical direct marker for nonadherence, as variations indicate erratic ingestion of medication. We previously reported that high variability in tacrolimus trough levels determined by the percent coefficient of variation (CV %) and standard deviation (SD) were associated with biopsy-proven rejection. We hypothesized that the CV % and SD in patients on a sirolimus/low-dose tacrolimus regimen may associate with self-reported medication nonadherence, rejection and donor-specific antibodies. In this pilot feasibility study, we studied 37 biopsies in 23 pediatric renal transplant patients on both sirolimus and tacrolimus immunosuppression; CV %, SD, de novo donor-specific antibodies, rejection, and self-reported adherence were examined. A cut-off sirolimus CV % of 25 maximized the percentage of biopsies correctly classified as rejection (32 of 37, or 86 %, p = 0.001). A cut-off tacrolimus CV % of 31 maximized the percentage of correctly classified biopsies (25 of 37, or 68 %, p = 0.09). Among patients with both high sirolimus and tacrolimus CV %, 67 % developed de novo donor-specific antibodies (p = 0.002) with a DQ predominance and 71 % reported nonadherence (p = 0.05). In pediatric renal transplantation, sirolimus and tacrolimus CV % is a potential tool for monitoring patients at risk for allograft rejection and donor-specific antibodies secondary to medication nonadherence.


Equine heart disease. An overview.
The clinician must appreciate unique aspects of equine cardiovascular physiology in order to distinguish normal variation from a pathophysiologic state. Cardiovascular problems in the horse include auscultation of cardiac murmurs, identification of cardiac arrhythmias, and recognition of congestive heart failure. A cardiac data base including history, general and cardiovascular examination, and resting and postexercise electrocardiogram are necessary to evaluate most horses for significant heart disease. Additional studies, such as echocardiography and cardiac catheterization, are useful in selected cases. Because some types of heart disease are amendable by therapy, an accurate diagnosis is needed to direct treatment and offer a realistic prognosis.


Neuroendocrine and haemodynamic responses to abdominal aortic cross clamp and release during high-dose opiate-oxygen-isoflurane anaesthesia.
The haemodynamic and neuroendocrine responses and beta adrenoceptor distribution associated with aortic cross clamping and release were quantified in 14 patients undergoing elective abdominal aortic aneurysm surgery using a high-dose opiate-oxygen-isoflurane anaesthetic technique. These changes were correlated with neutrophil beta adrenoceptor distribution. Aortic cross clamp application was associated with increased systemic vascular resistance (SVR), and decreased cardiac index (CI). Left ventricular stroke work index remained constant during the period of cross clamp application and following release. Cross clamp release was associated with increased CI and decreased SVR. Plasma cortisol concentrations did not change during the study period. Plasma catecholamine concentrations, although elevated prior to surgery, remained unchanged during aortic cross clamping and following release. The percentage of internalised beta adrenoceptors was elevated before surgery and was unaffected by surgical intervention. This study suggests that a high-dose fentanyl-oxygen-isoflurane anaesthetic technique in a patient population with high circulating catecholamine levels and downregulation of beta adrenoceptors is associated with cardiovascular stability and attenuated neuroendocrine responses.


Efficacy and safety of intermittent repeated levosimendan infusions in advanced heart failure patients: the LAICA study.
The aim of the LAICA study was to evaluate the long-term effectiveness and safety of intermittent levosimendan infusion in patients with advanced heart failure (AdHF). This was a multicentre, randomized, double-blind, placebo-controlled clinical trial of intermittent levosimendan 0.1 μg/kg/min as a continuous 24-h intravenous infusion administered once monthly for 1 year in patients with AdHF. The primary endpoint [incidence of rehospitalization (admission to the emergency department or hospital ward for >12 h) for acute decompensated HF or clinical deterioration of the underlying HF] occurred in 23/70 (33%) of the levosimendan group (Group I) and 12/27 (44%) of the placebo group (Group II) (P = 0.286). The incidence of hospital readmissions for acute decompensated HF (Group I vs. Group II) at 1, 3, 6, and 12 months was 4.2% vs. 18.2% (P = 0.036); 12.8% vs. 33.3% (P = 0.02); 25.7% vs. 40.7% (P = 0.147); 32.8% vs. 44.4% (P = 0.28), respectively. In a secondary pre-specified time-to-event analysis no differences were observed in admission for acute decompensated HF between patients treated with levosimendan compared with placebo (hazard ratio 0.66; 95% CI, 0.32-1.32; P = 0.24). Cumulative incidence for the aggregated endpoint of acute decompensation of HF and/or death at 1 and 3 months were significatively lower in the levosimendan group than in placebo group [5.7% vs. 25.9% (P = 0.004) and 17.1% vs. 48.1% (P = 0.001), respectively], but not at 6 and 12 months [34.2% vs. 59.2% (P = 0.025); 41.4% vs. 66.6% (P = 0.022), respectively]. Survival probability was significantly higher in patients who received levosimendan compared with those who received placebo (log rank: 4.06; P = 0.044). There were no clinically relevant differences in tolerability between levosimendan and placebo and no new safety signals were observed. In our study, intermittent levosimendan in patients with AdHF produced a statistically non-significant reduction in the incidence of hospital readmissions for acute decompensated HF, a significantly lower cumulative incidence of acute decompensation of HF and/or death at 1 and 3 month of treatment and a significant improvement in survival during 12 months of treatment.


Short-Term Total Hip Arthroplasty Outcomes in Patients With Psoriatic Arthritis or Psoriatic Skin Disease Compared to Patients With Osteoarthritis.
Outcomes of total hip arthroplasty (THA) in patients with psoriasis have been poorly studied. This study was undertaken to assess whether patients with psoriatic arthritis (PsA) or those with cutaneous psoriasis (PsC) without evidence of inflammatory joint disease are at an increased risk for worse outcomes after THA as compared to patients with osteoarthritis (OA). Among subjects in a prospective THA registry, PsA and PsC cases were identified by International Classification of Diseases, Ninth Revision codes, and all cases were matched with patients with OA as controls. Analyses were performed to identify predictors of poor postoperative pain or function. Of the 289 potential cases of PsA or PsC, 63 with PsA and 153 with PsC were validated. Self-report data were available postoperatively from 75% of PsA patients, 69% of PsC patients, and 94% of OA controls. In total, 51% of PsA patients and 56% of PsC patients were male, compared to 45% of OA controls (P = 0.04). Body mass index was higher in those with PsA or PsC (P = 0.002 versus controls). There were no differences in race or education between the 3 groups. PsA patients and PsC patients had more comorbidities than OA controls. PsA patients were more likely than PsC patients and OA controls to be current or previous smokers. Moreover, 54% of PsA patients were being treated with biologics or nonbiologic disease-modifying antirheumatic drugs, compared to 8% of PsC patients. There were no significant differences in pre- or postoperative Western Ontario and McMaster Universities OA Index scores for pain or function between the 3 groups. Short-Form 36 mental component summary scores were significantly better in the OA controls, both pre- and postoperatively (P = 0.006 and P < 0.001, respectively, versus PsA or PsC). EuroQol 5-domain health-related quality of life scores were significantly worse postoperatively for those with PsA or PsC (P < 0.0001 versus OA controls). In regression analyses, neither PsA nor PsC were risk factors for worse THA outcomes. Satisfaction with the outcomes of THA was similarly high among all 3 groups (P = 0.54). Neither PsA nor PsC are risk factors for poor outcomes after THA. This is important information to convey to patients with either PsA or PsC who are contemplating surgical intervention with THA.


Symptoms and reflux in infants: Infant Gastroesophageal Reflux Questionnaire Revised (I-GERQ-R)--utility for symptom tracking and diagnosis.
Answering a need for a thoroughly validated infant gastroesophageal reflux questionnaire, the Infant Gastroesophageal Reflux Questionnaire Revised (I-GERQ-R) was designed, refined, and validated using state-of-the-art psychometric methods. Diagnostic and evaluative (tracking) validity was identified. However, perplexing results of some clinical trials using the I-GERQ-R for diagnosis prompted analysis of possible reasons, including ambiguities in defining symptomatic gastroesophageal reflux disease (GERD) and aspects of the validation process. Symptomatic GERD is defined by "troublesomeness" of symptoms and attribution of their causation to reflux--two crucial issues. Methods of quantifying symptom-reflux associations are described and their limitations identified. The location of "symptomatic esophageal GERD" in the continuum of erosive GERD, histologic GERD, and nonerosive reflux disease is indicated, with the last including "suberosive," "premicroscopic," and "functional heartburn" subcategories. Another category is defined solely by surrogate measures of propensity to GERD (e.g., acid exposure thresholds defined on esophageal pH monitoring). During diagnostic validation of the Infant Gastroesophageal Reflux Questionnaire (I-GERQ) instruments, asymptomatic normals were contrasted with symptomatic GERD infants (who also tested positive with esophageal histology and esophageal pH monitoring). However, the diagnostic validation did not attempt to distinguish symptomatic GERD infants from symptomatic infants without GERD. The I-GERQ-R is thus adequately sensitive to be used diagnostically to screen infants for symptom burden, but should probably be supplemented by other, perhaps invasive, testing to assure appropriate specificity. The I-GERQ-R's validation for evaluative properties, however, supports its use for tracking symptoms within clinical trials.


The Potential Teratogenicity Alert for Women Conceiving on Dolutegravir-Based Regimens: An Assessment of Risk Communication by an Urban HIV Clinic in Uganda and Choices made by Women.
INTRODUCTION AND OBJECTIVE: In May 2018, the World Health Organization and other regulatory authorities released a safety alert for dolutegravir related to a risk of neural tube defects among women exposed to dolutegravir at the time of conception. Models of how drug safety information can be shared effectively in the shortest time are necessary to prevent interruptions of public health programs. We sought to describe an implementation process to inform and support women already on dolutegravir-based regimens at the time of conception to make informed choices following the safety alert of a potential teratogenicity risk. We describe the choices made by women, as well as determine the factors associated with women's choices to switch off dolutegravir. A clinic response plan was developed in the first week following the alert and clinic staff were trained on safety guidance. All women aged < 55 years taking dolutegravir were identified from the clinic database and contacted by phone for earlier appointments. Non-menopausal and non-surgically sterilized women were referred for urine pregnancy testing and evaluation of pregnancy intentions in the following 12 months and effective family planning was offered. We describe the coverage of women who received the communication as well as the fidelity to the outlined plan from 21 May to 12 September, 2018. We used modified a Poisson regression analysis to determine factors associated with switching off dolutegravir. Of all active patients in the clinic, 9% (690/7963) were identified as female aged < 55 years taking dolutegravir. Ninety-five percent (656/690) were reviewed by September 2018 and informed of the safety alert, implying a high level of uptake. Fidelity to standard operating procedures was also high at 72%. Twenty-two percent (146/656) of patients were menopausal or surgically sterilized. Five hundred and ten women were of reproductive potential with a median age (interquartile range) of 37 years (30-42 years). Five percent (23/510) were human chorionic gonadotrophin positive and all initial ultrasound reports revealed no deformities. Twenty-one percent (108/510) had intentions to conceive and opted to stop taking dolutegravir with 90% (97/108) switching to efavirenz. Seventy-nine percent (402/510) opted to remain taking dolutegravir. However, only 40% (160/402) chose effective contraceptive methods and 60% (242/402) opted for condoms only/no contraceptive method. A rapid well-coordinated response ensured prompt communication of the dolutegravir safety warning. The process developed by the clinic can act as a model for response during drug safety alerts. Women made informed decisions with most opting to remain taking dolutegravir; however, effective contraception uptake was low.


Strongyloides stercoralis: a rare cause of obstructive duodenal stenosis.
Strongyloidiasis is a rare helminthic infection in Europe, and it may cause duodenal obstruction. We report a patient who was admitted to our Medical Department with nausea and repeated vomiting since 10 years until food intake became impossible. Subsequent investigations revealed a duodenal obstruction at the upper third of the duodenum, as well as enterocolitis of the terminal ileum with eosinophils dispersed throughout the mucosa. Since food intake was still not possible after treatment with a course of i.v. PPI and prokinetic applications, we decided to perform a resection of the upper duodenum with Y-Roux reconstruction. The histopathological examination of the resected specimen revealed strongyloidiasis. Parasite infections such as strongyloidiasis represent a rare differential diagnosis of duodenal obstruction especially if patients originate from endemic regions.


[Treatment of peri-ocular skin lesions with the erbium:YAG laser].
A great number of consmetically disturbing, cutaneous lesions are localized in the periocular region. While various approaches for treatment such as excision, electrocauterization or cryosurgery often show unsatisfactory results, the use of laser technology is of increasing interest. A total of 104 patients with different cutaneous periorbital lesions (wrinkles, xanthelasma, syringoma) were treated with a new erbium: YAG laser system (wavelength 2940 nm, pulse duration 0.350 ms and pulse energy 0.1-1.7 Joules), which works on the principle of vaporization. The erbium: YAG laser allows athermic ablation of very thin skin layers without scarring and with minimal lateral thermal injury due an extremely short pulse duration. Periorbital lesions can be treated effectively by erbium: YAG laser, and good to excellent cosmetic results can be obtained. The use of pulsed erbium: YAG laser is an effective and promising new method for treatment of different superficial periorbital cutaneous lesions.


New opportunities for designing effective small interfering RNAs.
Small interfering RNAs (siRNAs) that silence genes of infectious diseases are potentially potent drugs. A continuing obstacle for siRNA-based drugs is how to improve their efficacy for adequate dosage. To overcome this obstacle, the interactions of antiviral siRNAs, tested in vivo, were computationally examined within the RNA-induced silencing complex (RISC). Thermodynamics data show that a persistent RISC cofactor is significantly more exothermic for effective antiviral siRNAs than their ineffective counterparts. Detailed inspection of viral RNA secondary structures reveals that effective antiviral siRNAs target hairpin or pseudoknot loops. These structures are critical for initial RISC interactions since they partially lack intramolecular complementary base pairing. Importing two temporary RISC cofactors from magnesium-rich hairpins and/or pseudoknots then kickstarts full RNA hybridization and hydrolysis. Current siRNA design guidelines are based on RNA primary sequence data. Herein, the thermodynamics of RISC cofactors and targeting magnesium-rich RNA secondary structures provide additional guidelines for improving siRNA design.


Mycobacterium abscessus pulmonary disease presenting with spontaneous pneumomediastinum and subcutaneous emphysema in childhood acute lymphoblastic leukemia: a case report and literature review.
Mycobacterium abscessus is a rapidly growing mycobacterium commonly identified in adults with underlying pulmonary diseases but is rarely observed in children. A better understanding of this pathogen in children is essential. We report the case of a 49-month-old female child without previous underlying pulmonary diseases but with acute lymphoblastic leukemia (ALL). The patient was complicated with pneumonia during chemotherapy, which was primarily characterized by spontaneous pneumomediastinum and subcutaneous emphysema on chest computed tomography (CT). M. abscessus sequences were detected by metagenomic next-generation sequencing in bronchoalveolar lavage fluid. With mechanical ventilation, closed thoracic drainage, and anti-infective therapy for 6 months, the patient's infection was controlled. The patient completed 2.5 years of treatment for ALL, and the drugs were discontinued. The patient currently remains in complete hematologic remission. We reviewed the literature on 33 children with M. abscessus pulmonary disease. These children mostly had underlying immunodeficiency. Chest CT most often showed nodular shadows, consolidation, and bronchiectasis. Spontaneous pneumomediastinum and subcutaneous emphysema were not reported as major manifestations. Spontaneous pneumomediastinum and subcutaneous emphysema were our patient's main characteristics on chest CT, and this study enriches the knowledge regarding possible imaging changes in M. abscessus pulmonary disease in children. This case report reflects good clinical experience in maintaining the balance between chemotherapy and anti-infective therapy in childhood ALL.


Ultrasonography in tarsal tunnel syndrome.
The purpose of this study was to clarify the diagnostic value of ultrasonography in tarsal tunnel syndrome. Seventeen patients (17 feet) with tarsal tunnel syndrome were treated between 1988 and 2003. Preoperative ultrasonography was performed, and the cause of the syndrome was confirmed intraoperatively in all cases. Long and short axes of the tarsal tunnel were scanned to ascertain the presence of any space-occupying lesion. The causes of tarsal tunnel syndrome, as confirmed by surgery, were ganglia (n = 10), talocalcaneal coalition (n = 1), talocalcaneal coalition associated with ganglia (n = 3), and varicose veins (n = 3). Among the cases involving ganglia, hypoechoic or anechoic regions were observed. The mean sizes +/- SD of these regions were 19.4 +/- 8.8 mm in the long axis, 15.2 +/- 6.3 mm in the short axis, and 10.4 +/- 3.8 mm in depth. Of these, 3 ganglia were not clearly palpable before surgery and were small: 10 x 10 x 7, 13 x 11 x 9, and 9 x 8 x 7 mm. Among the cases involving talocalcaneal coalition, ultrasonography indicated a beak-shaped bony process on the short axis images. Although these 3 cases were associated with ganglia, this could not be determined by preoperative palpation. As a diagnostic imaging technique for tarsal tunnel syndrome, ultrasonography is extremely useful for identifying space-occupying lesions. Ultrasonography should be performed routinely in patients with suspected tarsal tunnel syndrome.


Correlation of circulating dehydroepiandrosterone with activated protein C generation and carotid intima-media thickness in male patients with type 2 diabetes.
Dehydroepiandrosterone exerts a protective effect against cardiovascular diseases. However, the relationship of dehydroepiandrosterone with the anticoagulant factor activated protein C, generated by the thrombin-thrombomodulin complex on vascular endothelial cells, remains unknown. This study aimed at studying the relationship between dehydroepiandrosterone and activated protein C generation in patients with Type 2 diabetes. Sixty-two male patients with Type 2 diabetes were enrolled in this study. Data obtained from 40 healthy male subjects were used as controls. The plasma levels of dehydroepiandrosterone, the activated protein C-protein C inhibitor complex, high-sensitivity C-reactive protein and monocyte chemoattractant protein-1 were measured by enzyme immunoassays. Carotid intima-media thickness was measured by ultrasonography. The plasma levels of dehydroepiandrosterone (5.15 ± 2.81 vs. 3.76 ± 2.16 ng/ml; P < 0.005) and the activated protein C-protein C inhibitor complex (1.90 ± 1.07 vs. 1.02 ± 0.51 ng/ml; P < 0.001) were significantly lower in patients with diabetes than in normal subjects. Univariate analysis showed a significant correlation of the plasma level of dehydroepiandrosterone with that of the activated protein C-protein C inhibitor complex (r = 0.48, P < 0.001), high-sensitivity C-reactive protein (r = -0.30, P < 0.05) and with the mean intima-media thickness (r = -0.28, P < 0.05) in patients with diabetes. Stepwise multiple regression analysis showed that the plasma level of dehydroepiandrosterone is significantly correlated with the plasma levels of the activated protein C-protein C inhibitor complex (F = 18.06) and high-sensitivity C-reactive protein (F = 4.94). There was no correlation between the plasma levels of dehydroepiandrosterone and monocyte chemoattractant protein-1. These results suggest that lower circulating levels of dehydroepiandrosterone are associated with decreased activated protein C generation and higher intima-media thickness in patients with Type 2 diabetes.


The way things were and are. Changing indications for intraocular lens implantation.
In the midst of a rapidly exploding technology, guidelines for intraocular lens implantation in the United States were conservative several years ago in order to ensure good results and patient safety. Many of the views held then are now obsolete. The indications for its use should be left to the judgment of the surgeon and a properly informed patient. Intraocular lenses need no longer be restricted to the elderly, although caution is recommended in children and young adults. Restricting lens implants to one eye is obsolete, but there should be a reasonable time period of success before an implant is placed in the second eye. The approach to the one-eyed patient and those with other ocular pathology may now be modified.


mRNA, lncRNA, and circRNA expression profiles in a new aortic dissection murine model induced by hypoxia and Ang II.
Aortic dissection (AD) is a cardiovascular emergency with degeneration of the aortic media. Mounting evidence indicates obstructive sleep apnea (OSA) as an independent risk factor for AD development with unknown mechanisms. This study aims to establish a stable murine model of OSA-related AD (OSA-AD) and uncover the potential changes in gene transcripts in OSA-AD. ApoE-/- mice were exposed to the chronic intermittent hypoxia (CIH) system combined with Ang II administration to establish the OSA-AD model. Pathological staining was performed to exhibit the physiological structure of the mouse aorta. The SBC mouse ceRNA microarray was used to identify significantly differentially expressed (DE) mRNAs, DE long-non-coding RNAs (DElncRNAs), and DE circular RNAs (DEcircRNAs) in OSA-AD tissues. Subsequently, bioinformatics analysis, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genome (KEGG), and protein-protein interaction (PPI) analyses, were performed to evaluate the function of the significantly differentially expressed transcripts (DETs). The hub genes were confirmed using quantitative real-time polymerase chain reaction (qRT-PCR). ApoE-/- mice exposed to CIH and Ang II showed a high ratio of aortic accident (73.33%) and significant aortic diameter dilatation (1.96 ± 0.175 mm). A total of 1,742 mRNAs, 2,625 lncRNAs, and 537 circRNAs were identified as DETs (LogFC ≥ 1.5 or ≤ -1.5, P < 0.05). GO and KEGG analyses demonstrated that the differentially expressed mRNAs (DEmRNAs) were most enriched in cell proliferation, migration, apoptosis, inflammation, and hypoxia-related terms, which are closely related to aortic structural homeostasis. The PPI network contained 609 nodes and 934 connections, the hub genes were highlighted with the CytoHubba plugin and confirmed by qRT-PCR in AD tissues. KEGG pathway analysis revealed that the cis-regulated genes of DElncRNAs and circRNAs-host genes were enriched in aortic structural homeostasis-related pathways. Our findings help establish a de novo OSA-AD animal model using ApoE-/- mice. Many DEmRNAs, DElncRNAs, and DEcircRNAs were screened for the first time in OSA-AD tissues. Our findings provide useful bioinformatics data for understanding the molecular mechanism of OSA-AD and developing potential therapeutic strategies for OSA-AD.


The effect of pore diameter on neo-tissue formation in electrospun biodegradable tissue-engineered arterial grafts in a large animal model.
To date, there has been little investigation of biodegradable tissue engineered arterial grafts (TEAG) using clinically relevant large animal models. The purpose of this study is to explore how pore size of electrospun scaffolds can be used to balance neoarterial tissue formation with graft structural integrity under arterial environmental conditions throughout the remodeling process. TEAGs were created with an outer poly-ε-caprolactone (PCL) electrospun layer and an inner sponge layer composed of heparin conjugated 50:50 poly (l-lactide-co-ε-caprolactone) copolymer (PLCL). Outer electrospun layers were created with four different pore diameters (4, 7, 10, and 15 µm). Fourteen adult female sheep underwent bilateral carotid artery interposition grafting (n = 3-4 /group). Our heparin-eluting TEAG was implanted on one side (n = 14) and ePTFE graft (n = 3) or non-heparin-eluting TEAG (n = 5) on the other side. Twelve of the fourteen animals survived to the designated endpoint at 8 weeks, and one animal with 4 µm pore diameter graft was followed to 1 year. All heparin-eluting TEAGs were patent, but those with pore diameters larger than 4 µm began to dilate at week 4. Only scaffolds with a pore diameter of 4 µm resisted dilation and could do so for up to 1 year. At 8 weeks, the 10 µm pore graft had the highest density of cells in the electrospun layer and macrophages were the primary cell type present. This study highlights challenges in designing bioabsorbable TEAGs for the arterial environment in a large animal model. While larger pore diameter TEAGs promoted cell infiltration, neotissue could not regenerate rapidly enough to provide sufficient mechanical strength required to resist dilation. Future studies will be focused on evaluating a smaller pore design to better understand long-term remodeling and determine feasibility for clinical use. STATEMENT OF SIGNIFICANCE: In situ vascular tissue engineering relies on a biodegradable scaffold that encourages tissue regeneration and maintains mechanical integrity until the neotissue can bear the load. Species-specific differences in tissue regeneration and larger mechanical forces often result in graft failure when scaling up from small to large animal models. This study utilizes a slow-degrading electrospun PCL sheath to reinforce a tissue engineered arterials graft. Pore size, a property critical to tissue regeneration, was controlled by changing PCL fiber diameter and the resulting effects of these properties on neotissue formation and graft durability was evaluated. This study is among few to report the effect of pore size on vascular neotissue formation in a large animal arterial model and also demonstrate robust neotissue formation.


Intermittent dietary methionine deprivation facilitates tumoral ferroptosis and synergizes with checkpoint blockade.
Dietary methionine interventions are beneficial to apoptosis-inducing chemotherapy and radiotherapy for cancer, while their effects on ferroptosis-targeting therapy and immunotherapy are unknown. Here we show the length of time methionine deprivation affects tumoral ferroptosis differently. Prolonged methionine deprivation prevents glutathione (GSH) depletion from exceeding the death threshold by blocking cation transport regulator homolog 1 (CHAC1) protein synthesis. Whereas, short-term methionine starvation accelerates ferroptosis by stimulating CHAC1 transcription. In vivo, dietary methionine with intermittent but not sustained deprivation augments tumoral ferroptosis. Intermittent methionine deprivation also sensitizes tumor cells against CD8+ T cell-mediated cytotoxicity and synergize checkpoint blockade therapy by CHAC1 upregulation. Clinically, tumor CHAC1 correlates with clinical benefits and improved survival in cancer patients treated with checkpoint blockades. Lastly, the triple combination of methionine intermittent deprivation, system xc- inhibitor and PD-1 blockade shows superior antitumor efficacy. Thus, intermittent methionine deprivation is a promising regimen to target ferroptosis and augment cancer immunotherapy.


Evaluation of surveillance for surgical site infections in Thika Hospital, Kenya.
In low-income countries, surgical site infections (SSIs) are a very frequent form of hospital-acquired infection. Surveillance is an important method for controlling SSI but it is unclear how this can best be performed in low-income settings. To examine the epidemiological characteristics of various components of an SSI surveillance programme in a single Kenyan hospital. The study assessed the inter-observer consistency of the surgical wound class (SWC) and American Society of Anesthesiologists (ASA) scores using the kappa statistic. Post-discharge telephone calls were evaluated against an outpatient clinician review 'gold standard'. The predictive value of components of the Centers for Disease Control and Prevention - National Healthcare Safety Network (CDC-NHNS) risk index was examined in patients having major obstetric or gynaecological surgery (O&G) between August 2010 and February 2011. After appropriate training, surgeons and anaesthetists were found to be consistent in their use of the SWC and ASA scores respectively. Telephone calls were found to have a sensitivity of 70% [95% confidence interval (CI): 47-87] and a specificity of 100% (95% CI: 95-100) for detection of post-discharge SSI in this setting. In 954 patients undergoing major O&G operations, the SWC score was the only parameter in the CDC-NHNS risk index model associated with the risk of SSI (odds ratio: 4.00; 95% CI: 1.21-13.2; P = 0.02). Surveillance for SSI can be conducted in a low-income hospital setting, although dedicated staff, intensive training and local modifications to surveillance methods are necessary. Surveillance for post-discharge SSI using telephone calls is imperfect but provides a practical alternative to clinic-based diagnosis. The SWC score was the only predictor of SSI risk in O&G surgery in this context.


Is slow follicular growth the cause of silent estrus in water buffaloes?
The present experiment was conducted to study the growth profile of the ovulatory follicle in relation to the expression of estrus following administration of PGF(2alpha) to subestrus buffaloes. After detection of a mature corpus luteum by examination per rectum, confirmed by ultrasound scanning, subestrus buffaloes (n=20) were treated (Day 0) with single dose of Dinoprost tromethamin (25 mg, i.m.). Blood samples were collected at 0, 24 and 48 h after treatment for estimation of plasma progesterone concentration. Growth profile of the ovulatory follicle was monitored daily through ultrasound scanning starting from Day 0 until ovulation and the regression profile of CL was monitored at 0, 24 and 48 h of treatment. Estrus was detected by exposure to a fertile buffalo bull three times a day until expression of overt estrus or ovulation. Behavioral estrus was recorded in 14 animals and 6 animals ovulated silently. Sixteen animals including six animals with silent estrus ovulated from the dominant follicle present at treatment (Group A) and remaining four animals ovulated from the dominant follicle of succeeding follicular wave (Group B). The intervals from treatment to estrus (6.5+/-0.25 versus 3.2+/-0.27 days, P<0.001) and treatment to ovulation (7.5+/-0.25 versus 5.4+/-0.46 days, P<0.005) were significantly longer in animals of Group B compared with animals of Group A. Significant differences were observed in growth profile of the ovulatory follicle between animals of Groups A and B with respect to size of the follicle on Day 0 (9.8+/-0.7 versus 5.3+/-0.45 mm, P<0.001), daily growth rate (0.97+/-0.07 versus 1.6+/-0.2 mm/day, P<0.01) and increase in diameter (4.1+/-0.6 versus 7.8+/-0.7 mm, P<0.01). The animals with silent estrus (subgroup A-2) had significantly smaller diameter of the ovulatory follicle on Day 0 (7.7+/-0.4 versus 11.0+/-0.7 mm, P<0.005), its daily growth rate was significantly slower (0.7+/-0.02 versus 1.1+/-0.1 mm/day, P<0.01) and they recorded significantly longer interval from treatment to ovulation (7.3+/-0.56 versus 4.2+/-0.27 days, P<0.001) compared with the animals that showed overt estrus (subgroup A-1). The corpus luteum area (CL area) and plasma progesterone (P(4)) concentration declined continuously from 0 to 48 h after PGF(2alpha) treatment in the animals of both the Groups A and B. Non-significant differences were observed in mean CL area and plasma P(4) concentration at 0, 24 and 48 h post-treatment between animals of Groups A and B and also between animals of subgroups A-1 and A-2. The small size and the slow growth rate of the ovulatory follicle were identified as the possible cause of silent estrus in subestrus buffaloes after PGF(2alpha) treatment.


Measles seroprevalence among Dutch travelling families.
While measles vaccination is widely implemented in national immunisation programmes, measles incidence rates are increasing worldwide. Dutch inhabitants who were born between 1965-1975 may have fallen between two stools, lacking protection from a natural infection, and having missed the introduction of the measles vaccination schedule. With this study we aim to find the measles seroprevalence in travellers born between 1965 and 1975, compared to those born before 1965 and after 1975. Families travelling to Eastern Europe or outside Europe during the preceding year were recruited via Dutch secondary schools between 2016 and 2018. Their vaccination status was assessed using questionnaires, vaccination records and measles serology in dried blood spot (DBS) eluates. Measles virus antibody concentrations were determined with an ELISA (EUROIMMUNE®) and a subset was retested with a focus reduction neutralization assay (FRNT). In 188 (79%) of the 239 available DBS eluates, the ELISA could detect sufficient measles virus-specific IgG antibodies. Of the negative samples that were retested with FRNT, 85% remained negative, resulting in an overall seroprevalence of 82% [95% CI 76-86]. Children had a lower seroprevalence (72%) than adults (87%). Travellers born between 1965 and 1975 were protected in 89%. In this study, we report a measles seroprevalence of 82% among Dutch travelling families. Remarkably, seroprevalence rates were lowest in children (12-18 years) instead of travellers born between 1965 and 1975. Although a fraction of people without detectable antibodies may be protected by other immune mechanisms, these data suggest that measles (re)vaccination should be considered for travellers to endemic regions.


Myotonic dystrophy is closely linked to the gene for muscle-type creatine kinase (CKMM).
We have studied genetic linkage between the gene for creatine kinase muscle type (CKMM) and the gene for myotonic dystrophy (DM). In a panel of 65 myotonic dystrophy families from Canada and the Netherlands, a maximum lod score (Zmax) of 22.8 at a recombination frequency (theta) of 0.03 was obtained. Tight linkage was also demonstrated for CKMM and the gene for apolipoprotein C2 (ApoC2). This establishes CKMM as a useful marker for myotonic dystrophy.


[Cell suspension culture and flavonoids production in Saussurea involucrate].
To establish a system of cell suspension culture for Saussurea involucrate. The effects of different factors on cell growth and flavonoids production of S. involucrate were systematically studied including the media, initial pH values of the medium, carbon sources, inoculum quantity, and plant growth substance. The optimum medium was N6, initial pH values of the medium was 5.8, sucrose concentration was 50 g x L(-1), inoculum quantity was 60-80 g x L(-1) FW. Medium supplemented with BA (0.5 mg x L(-1)) and NAA (3 mg x L(-1)) was suitable for cell growth, but medium containing BA (0.2 mg x L(-1)) and NAA (2 mg x L(-1)) was suitable for flavonoids production. Cell growth and flavonoids production in the suspention of S. involucrate culture cell should be optimized.


Effect of kernel breakage on the fermentation profile, nitrogen fractions, and in vitro starch digestibility of whole-plant corn silage and ensiled corn grain.
The objective of this experiment was to analyze the effect of kernel breakage on the fermentation profile, nitrogen fractions, and ruminal in vitro starch digestibility of whole-plant corn silage and ensiled corn grain. Whole corn plants were harvested, and ears were separated from the forage portion and shelled. Corn kernels were either left intact or broken manually using a hammer. The remaining forage portion of the corn plants was chopped. Samples of the intact and broken kernels were stored for 0 or 30 d in quadruplicate vacuum pouches. Remaining intact and broken kernels were each reconstituted with the chopped forage portion of the corn plant to simulate whole-plant corn forage and were also stored for 0 or 30 d. In kernels separated from whole-plant corn silage, kernel form had no effect on zein protein concentrations. However, it was observed that in vitro starch digestibility at 7 h increased with ensiling only in kernels that were broken. When corn kernels were ensiled alone, concentrations of soluble crude protein and ammonia nitrogen increased with ensiling to a greater extent when kernels were broken. Finally, fermentation of ensiled corn grain was enhanced when kernels were broken. Overall, this study gives insight into the importance of kernel breakage to improve starch digestibility in corn silage through means other than a reduction in particle size and opens the door for continued investigation into the proteolytic activity occurring in the silo.


The epidemiology and impact of pain in osteoarthritis.
Osteoarthritis (OA) is the most common form of arthritis and a leading cause of disability worldwide, largely due to pain, the primary symptom of the disease. The pain experience in knee OA in particular is well-recognized as typically transitioning from intermittent weight-bearing pain to a more persistent, chronic pain. Methods to validly assess pain in OA studies have been developed to address the complex nature of the pain experience. The etiology of pain in OA is recognized to be multifactorial, with both intra-articular and extra-articular risk factors. Nonetheless, greater insights are needed into pain mechanisms in OA to enable rational mechanism-based management of pain. Consequences of pain related to OA contribute to a substantial socioeconomic burden.


Diagnostic Performance of Dynamic Contrast-Enhanced MRI and 18F-FDG PET/CT for Evaluation of Soft Tissue Tumors and Correlation with Pathology Parameters.
To assess the diagnostic performance of dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) and fluorine-18-fluorodeoxyglucose (18F-FDG) positron-emission tomography/computed tomography (PET/CT) parameters in evaluating the biological behavior of soft tissue tumors. We retrospectively analyzed DCE-MRI and 18F-FDG PET/CT parameters in 78 patients with pathology-confirmed soft tissue tumors. A total of 78 patients had undergone DCE-MRI examination, while 24 patients with malignant soft tissue tumor had undergone 18F-FDG PET/CT examination. Microvessel density (MVD) and the Ki-67 labeling index (LI) were detected using immunohistochemistry. Differences in parameters (Ktrans, Kep, Ve, MVD, and Ki-67 LI) between benign and malignant tumors were compared. Differences in parameters (Ktrans, Kep, Ve, MVD, and SUVmax) between high- and low-proliferation malignant tumors (grouped by Ki-67 LI) were compared. Correlation of the DCE-MRI and 18F-FDG PET/CT parameters with MVD and Ki-67 LI was analyzed. Only the Ktrans, Kep, MVD, and Ki-67 LI differed significantly between the benign and malignant soft tissue tumors (all p < 0.001). Only Kep (p = 0.033) and SUVmax (p = 0.001) differed significantly between high- and low-proliferation malignant soft tissue tumors. Ktrans, Kep, and SUVmax correlated positively with MVD (r = 0.805, 0.778, 0.730, respectively; all p < 0.001), and with Ki-67 LI (r = 0.721, 0.685, 0.655, respectively; all p < 0.001). DCE-MRI and 18F-FDG PET/CT parameters indicate soft tissue tumor biological behavior and can be used to differentiate between benign and malignant soft tissue tumors and between high- and low-proliferation malignant soft tissue tumors.


Side effects of low-dose tamoxifen: results from a six-armed randomised controlled trial in healthy women.
Adherence to adjuvant tamoxifen therapy is suboptimal, and acceptance of tamoxifen for primary prevention is poor. Published results indicate effect of low-dose tamoxifen therapy. Using questionnaire data from a randomised controlled trial, we describe side effects of standard and low-dose tamoxifen in healthy women. In the KARISMA trial, 1440 healthy women were randomised to 6 months of daily intake of 20, 10, 5, 2.5, 1 mg of tamoxifen or placebo. Participants completed a 48-item, five-graded Likert score symptom questionnaire at baseline and follow-up. Linear regression models were used to identify significant changes in severity levels across doses and by menopausal status. Out of 48 predefined symptoms, five were associated with tamoxifen exposure (hot flashes, night sweats, cold sweats, vaginal discharge and muscle cramps). When comparing these side effects in premenopausal women randomised to low doses (2.5, 5 mg) versus high doses (10, 20 mg), the mean change was 34% lower in the low-dose group. No dose-dependent difference was seen in postmenopausal women. Symptoms related to tamoxifen therapy are influenced by menopausal status. Low-dose tamoxifen, in contrast to high-dose, was associated with less pronounced side effects, a finding restricted to premenopausal women. Our findings give new insights which may influence future dosing strategies of tamoxifen in both the adjuvant and preventive settings. ClinicalTrials.gov ID: NCT03346200.


Aplastic anemia: a common hematological abnormality among peripheral pancytopenia.
Aplastic anemia is a well-recognized form of marrow failure. The incidence of aplastic anemia is subjected to wide variation. Most cases are acquired and immune-mediated but there are also inherited forms. The study was conducted to assess the magnitude of the problem, morphological changes and determinants of aplastic anemia in North Bengal. A cross-sectional study had been conducted for a period of one year among 5 to 70 years age group. Initially complete blood count followed by bone marrow examination was done for diagnosis. Out of 48 cases, 38 (79.17%) had hypocellular diagnosed as aplastic anemia, 5 (10.42%) each had normocellular and hypercellular bone marrow. Histopathology shows that 30 (78.95%) cases had increased iron stores in bone marrow and 8 (21.05%) cases had decreased iron stores. Subjects less than 20 years of age mostly (31.58%) suffered from non-severe disease in contrast to subjects in higher age groups who had severe or very severe disease, though the trend was not significantly different (χ(2) for linear trend 0.171, P > 0.05). The study shows aplastic anemia is a common hematological abnormality among peripheral pancytopenia in North Bengal region where males were affected more than females.


On the nature of the chemical noise in MALDI mass spectra.
The so-called "chemical noise background" imposes a major limit on the practical sensitivity of MALDI mass spectrometry. Typically, as the amount of material of interest subjected to MALDI analysis is reduced, the signal decreases to the point where it can no longer be differentiated from the chemical noise. Using a newly designed MALDI-ion trap mass spectrometer, we describe experiments intended to throw light on the nature of the chemical noise background and to reduce its effects. Single-stage mass spectrometric signals from peptides were observed to disappear into the noise when the amount of sample applied to the MALDI sample stage was decreased to less than a femtomole. At these low levels, analysis of the collision-induced fragmentation spectra revealed the presence of ions originating from the peptide as well as cluster ions that originate from the chemical noise. The fragmentation pattern arising from dissociation of the cluster species suggests that they are composed largely of matrix molecules. A significant fraction of these cluster ions can be dissociated at activation energies lower than the threshold for peptide fragmentation. We used this finding to collisionally pre-activate MALDI ions to remove a significant portion of the chemical noise from the spectrum, allowing us to obtain readily discernible single stage MS signals from 100 attomols of peptide. The strategy also yielded high quality MS/MS spectra from 100 attomols of peptide. Different possibilities of collisional pre-activation for improving sensitivity are considered.


Cortisol metabolism by human liver in vitro--IV. Metabolism of 9 alpha-fluorocortisol by human liver microsomes and cytosol.
The oxidative and reductive biotransformations of 9 alpha-fluorocortisol (fluorocortisol) by human liver microsomes and cytosol have been characterized. 9 alpha-Fluorination greatly simplified cortisol metabolism in microsomes: dehydrogenation of the 11 beta-hydroxyl group and A-ring (4-ene-5 beta and 3 alpha-keto) reduction, the principle pathways, were completely blocked. Fluorocortisol was essentially metabolized by the remaining pathways, 20 beta-reduction and 6 beta-hydroxylation. In cytosol, 20 beta-reduction replaced the A-ring reduction of cortisol as the sole biotransformation. The major structure-metabolism relationships of fluorocortisol in man, i.e. complete and extensive inhibition of 11 beta-dehydrogenation and 4-ene-5 beta-reduction, respectively, were attributed to hepatic enzyme systems. Their mechanistic basis is discussed with reference to the electronic and conformational changes induced by 9 alpha-fluorination.


Modulatory effects of phosphatidylserine on the binding of muscarinic cholinergic receptor ligands. Studies in vitro and in vivo.
The modulation of the binding of muscarinic cholinergic receptor ligands by phosphatidylserine purified from bovine cerebral cortex (BC-PS) was examined in vitro and in vivo. The enrichment of bovine cerebral cortical synaptosomal membranes with BC-PS, using a fusion technique, produced a concentration-dependent decrease in the affinity (increase in Kd) of [3H]quinuclidinyl benzylate (3H-QNB) specific binding to muscarinic acetylcholine receptors (mAChR), without changes in their maximal number (Bmax). Similar results were observed when [3H]oxotremorine (3H-OXO) was used to label a high affinity subpopulation of mAChR. On the other hand, preincubation of BC-PS liposomes with synaptosomal membranes in a nonoptimum fusion condition (at pH 7.4) did not alter the binding properties of both radioligands. Fusion experiments using a pure phosphatidylserine preparation from spinal cord revealed a similar decrement in the affinity of 3H-QNB specific binding. Five day's intraperitoneal (i.p.) administration of 15 mg/kg of BC-PS liposomes in rats increased the maximal number of cerebral cortical binding sites for 3H-OXO. Scatchard analysis revealed no changes in the apparent dissociation constant. This modification is selective in relation to the neural structure studied. Thus, BC-PS treatment did not modify 3H-OXO binding in the hippocampal formation and cerebellum. In contrast, parallel experiments using the muscarinic antagonist 3H-QNB showed no alteration in the binding properties of mAChR. Five day's i.p. administration of 15 mg/kg/d of phosphatidylcholine from bovine cerebral cortex (BC-PC) liposomes produced quite similar results to those obtained with BC-PS. These results indicate that mAChR are under the modulatory action of phosphatidylserine (PS) and phosphatidylcholine (PC), and suggest that this endogenous phospholipids may play a regulatory role on the mAChR. The possible implications of these findings on the effects of PC or PS treatment in neurological disorders involving a decrease in central cholinergic functions are discussed.


Quantitative evaluation of the outcomes of revascularization procedures for peripheral arterial disease using indocyanine green angiography.
We performed indocyanine green angiography (ICGA) in patients with peripheral arterial disease (PAD), and established a method for the quantitative measurement of appropriate parameters to assess peripheral perfusion and the applicability of ICGA tests. Twenty-one patients with PAD underwent revascularization procedures with pre- and postinterventional ICGA tests. The ICGA parameters, which included the magnitude of intensity of indocyanine green, the time to maximum intensity, and the time from fluorescence onset to half the maximum intensity (T1/2) were compared with the ankle-brachial pressure index, toe -brachial pressure index, and toe pressure. We evaluated these parameters for regions of interest (ROIs). T1/2 was the strongest parameter among all parameters of the ICGA tests. ROI 3, which included the distal region of the first metatarsal bone, correlated more significantly with the traditional measurements than the other ROIs. A value of T1/2 >20 seconds for ROI 3 was significantly correlated with a toe pressure of <50 mmHg (sensitivity: 0.77, specificity: 0.80). ICGA can be used to assess peripheral tissue perfusion. By measuring the value of T1/2 in ROI 3, ICGA tests can be used to evaluate the outcomes of revascularization procedures.