[Hyponatremia in a 58-year-old female patient with EGFR-positive lung adenocarcinoma].
The case of a 58-year old female patient with epidermal growth factor-positive pulmonary adenocarcinoma treated with the tyrosine kinase inhibitor afatinib is reported. After several months of first-line therapy the patient developed severe hyponatremia and tumor reassessment revealed a progressive course of the lung cancer. Rebiopsy showed transformation of the tumor into small-cell lung cancer. Therapy with afatinib was stopped immediately and platin-based chemotherapy was started. This case shows that tumor transformation under tyrosine kinase inhibitor therapy from non-small-cell into small-cell lung cancer can occur in rare cases.


Intraoperative radiotherapy in the multimodality approach to colorectal cancer.
The addition of intraoperative radiotherapy (IORT) to the multimodality approach for the treatment of locally advanced and locally recurrent colorectal cancer seems to result in improvements in local control and long-term survival. Local control and survival are most likely in patients in whom a gross total resection is accomplished. Peripheral nerve is the dose-limiting structure for patients treated with IORT. Further improvements in local control require the addition of dose modifiers during external beam radiotherapy or IORT. Distant relapse remains problematic, and effective systemic therapy is necessary to significantly improve long-term survival.


Comprehensive analysis of circRNA-miRNA-mRNA in oral squamous cell carcinoma.
The aim of the present study was to explore expression profiles of circular RNAs (circRNAs) and potential molecular mechanisms in oral squamous cell carcinoma (OSCC). In this study, high-throughput microarray was used to detect the expression profiles of circRNAs in OSCC. Reverse transcription quantitative PCR (RT-qPCR) was used to quantify the expression of dysregulated circRNAs in microarray. The validated microRNAs sponged by circRNAs and targeted mRNA were collected from literatures and miRTarBase database. and Gene Ontologypathways were analyzed on DAVID platform. Furthermore, network of circRNA-miRNA-mRNA and protein-protein interaction were constructed by Cytoscape 6.0 software. The results revealed that 1540 circRNAs were differentially expressed in 4 paired OSCC tissues and adjacent tissues significantly. Among them, 6 abundant circRNAs which have been reported in literatures were identified through RT-qPCR. According to the hypothesis of competing endogenous RNAs, 14 miRNAs sponged by the validated circRNAs and 62 downstream target protein coding genes were identified. Upon Gene Ontologyanalysis, 62 target genes were found to play roles in positively regulation of cell migration, cell division, angiogenesis and response to hypoxia. For Kyoto Encyclopedia of Genes and Genomes pathways, target genes were found to be involved in oncogenic signaling pathways (hsa05200), PI3K-Akt signaling pathway (hsa04151), focal adhesion (hsa04510), microRNAs in cancer (hsa05206), ECM-receptor interaction (hsa04512). These results suggested that circRNAs may participate in the tumorigenesis and development of OSCC via sponging miRNAs. This study indicated that circRNA may play vital role in tumorigenesis and metastasis in OSCC.


Reuse of carboplatin and paclitaxel in patients with relapsed endometrial cancer--the British Columbia Cancer Agency experience.
To evaluate the efficacy of reusing carboplatin and taxol in women with relapsed endometrial cancer. Retrospective analysis of our database of newly diagnosed high-risk patients with endometrial cancer treated with carboplatin-paclitaxel at diagnosis, with subsequent relapse for the period of 1995-2007. 111 patients of 200 relapsed. They had either endometroid or papillary serous histologies. Strategies utilized upon first relapse were: no treatment (n=33), surgery (n=4), hormones (n=8), irradiation (n=14) and chemotherapy (n=52). Carboplatin and paclitaxel was reused in 31 (60% of 52 retreated with chemotherapy or 29% of the total cohort of 111). There was no statistically significant difference in stage at diagnosis or grade at diagnosis between those retreated with chemotherapy or not or with carboplatin-paclitaxel versus another regimen. The patients retreated were a selected subgroup as only those with initial response or treated adjuvantly were offered carboplatin-paclitaxel. CR or PR were achieved in 8 (42%) patients with endometroid type cancer. In the papillary serous group 6 (50%) had CR or PR. Median PFS from first relapse was 8 months for endometroid and 9 months for papillary serous histology. OS was 15 months and 26 months respectively from first relapse. Carboplatin-taxol regimen is an efficacious treatment. Due to the patient selection these outcomes reported are likely to be an overstatement of what could be achieved in practice.


Sequence variability analysis of human class I and class II MHC molecules: functional and structural correlates of amino acid polymorphisms.
Major histocompatibility complex class I (MHCI) and class II (MHCII) molecules display peptides on antigen-presenting cell surfaces for subsequent T-cell recognition. Within the human population, allelic variation among the classical MHCI and II gene products is the basis for differential peptide binding, thymic repertoire bias and allograft rejection. While available 3D structural analysis suggests that polymorphisms are found primarily within the peptide-binding site, a broader informatic approach pinpointing functional polymorphisms relevant for immune recognition is currently lacking. To this end, we have now analyzed known human class I (774) and class II (485) alleles at each amino acid position using a variability metric (V). Polymorphisms (V>1) have been identified in residues that contact the peptide and/or T-cell receptor (TCR). Using sequence logos to investigate TCR contact sites on HLA molecules, we have identified conserved MHCI residues distinct from those of conserved MHCII residues. In addition, specific class II (HLA-DP, -DQ, -DR) and class I (HLA-A, -B, -C) contacts for TCR binding are revealed. We discuss these findings in the context of TCR restriction and alloreactivity.


Exclusive percutaneous approach for surgical transaortic transcatheter valve replacement.
Direct transaortic implantation (TAo) has been described as a new alternative approach for transcatheter aortic valve implantation in patients with unsuitable transfemoral access. TAo is usually achieved through an upper ministernotomy or, more recently, through a right thoracotomy in the second intercostal space. We describe here our first experience with a fully thoracoscopic approach that allowed successful deployment of a 23-mm Edwards SAPIEN valve.


Enterococci as probiotics and their implications in food safety.
Enterococci belong to the lactic acid bacteria (LAB) and they are of importance in foods due to their involvement in food spoilage and fermentations, as well as their utilisation as probiotics in humans and slaughter animals. However, they are also important nosocomial pathogens that cause bacteraemia, endocarditis and other infections. Some strains are resistant to many antibiotics and possess virulence factors such as adhesins, invasins, pili and haemolysin. The role of enterococci in disease has raised questions on their safety for use in foods or as probiotics. Studies on the incidence of virulence traits among enterococcal strains isolated from food showed that some can harbour virulence traits, but it is also thought that virulence is not the result of the presence of specific virulence determinants alone, but is rather a more intricate process. Specific genetic lineages of hospital-adapted strains have emerged, such as E. faecium clonal complex (CC) 17 and E. faecalis CC2, CC9, CC28 and CC40, which are high risk enterococcal clonal complexes. These are characterised by the presence of antibiotic resistance determinants and/or virulence factors, often located on pathogenicity islands or plasmids. Mobile genetic elements thus are considered to play a major role in the establishment of problematic lineages. Although enterococci occur in high numbers in certain types of fermented cheeses and sausages, they are not deliberately added as starter cultures. Some E. faecium and E. faecalis strains are used as probiotics and are ingested in high numbers, generally in the form of pharmaceutical preparations. Such probiotics are administered to treat diarrhoea, antibiotic-associated diarrhoea or irritable bowel syndrome, to lower cholesterol levels or to improve host immunity. In animals, enterococcal probiotics are mainly used to treat or prevent diarrhoea, for immune stimulation or to improve growth. From a food microbiological point of view, the safety of the bacteria used as probiotics must be assured, and data on the major strains in use so far indicate that they are safe. The advantage of use of probiotics in slaughter animals, from a food microbiological point of view, lies in the reduction of zoonotic pathogens in the gastrointestinal tract of animals which prevents the transmission of these pathogens via food. The use of enterococcal probiotics should, in view of the development of problematic lineages and the potential for gene transfer in the gastrointestinal tract of both humans and animals, be carefully monitored, and the advantages of using these and new strains should be considered in a well contemplated risk/benefit analysis.


Hunger-satiety signals in patients with Graves' thyrotoxicosis before, during, and after long-term pharmacological treatment.
Patients with Graves' thyrotoxicosis lose weight despite increased appetite and food intake, thus suggesting a disturbed balance between energy intake and expenditure. Underlying mechanisms are not fully elucidated. The objective of this study was to investigate whether hormonal factors, known to affect hunger/satiety, change significantly over time as pharmacological treatment turns hyperthyroidism into euthyroidism. For that purpose 11 patients with Graves' thyrotoxicosis were given thiamazole and l-thyroxine for 18-20 mo. They were investigated on three occasions: Test 1: before pharmacological therapy; Test 2: during medication; Test 3: a few months after conclusion of the pharmacological treatment. Sixteen healthy subjects were also investigated for comparison. The participants were fasted overnight. Blood samples for determination of plasma glucose and serum concentrations of free T3 and T4, TSH, albumin, cortisol, insulin, GH, IGF-1, IGFBP-1, leptin, and ghrelin were drawn in the morning from an antecubital vein. Laboratory data obtained in test 1 were statistically compared with those in tests 2 and 3. The study showed that the free T3 level declined from 42.8 +/- 4.3 pmol/L in test 1 to 6.0 +/- 0.8 pmol/L in test 2 (85 +/- 2% decline), and 5.5 +/- 0.3 pmol/L in test 3 (86 +/- 2% decline). The free T4 level fell concomitantly from 65.2 +/- 4.8 to 16.6 +/- 1.7 and 14.4 +/- 1.2 pmol/L. The glucose level was significantly higher during hyperthyroidism (test 1) than during euthyroidism (tests 2 and 3), whereas cortisol, insulin, GH, IGF-1, and leptin levels were similar. The IGFBP-1 level was initially high (48.8 +/- 8.5 microg/L in test 1), but with a relative decline in free T3 of 85 +/- 2% (test 2) the IGFBP-1 level declined by 34 +/- 13%, and with a free T3 decline of 86 +/- 2% (test 3) the binding protein fell by 39 +/- 29%. This brought about increased IGF-1 bioavailability as reflected by a rising IGF-1/IGFBP-1 ratio from 5.1 +/- 1.1 to 13.8 +/- 2.9 (p < 0.01). The ghrelin level was low (2454 +/- 304 ng/L) in test 1. It increased to 3127 +/- 397 ng/L in test 2 (p < 0.05), and to 3348 +/- 279 ng/L in test 3 (p < 0.01). Both ghrelin secretion and IGF-1 bioavailability are low in patients with untreated thyrotoxicosis, but increase markedly as pharmacotherapy makes them euthyroid. These metabolic changes may be caused by the transition of hyperthyroidism into euthyroidism rather than by the pharmacotherapy per se, since the metabolic changes prevailed also in the posttreatment period.


COMPASS: a formal framework and aggregate dataset for generalized surgical procedure modeling.
We propose a formal framework for the modeling and segmentation of minimally invasive surgical tasks using a unified set of motion primitives (MPs) to enable more objective labeling and the aggregation of different datasets. We model dry-lab surgical tasks as finite state machines, representing how the execution of MPs as the basic surgical actions results in the change of surgical context, which characterizes the physical interactions among tools and objects in the surgical environment. We develop methods for labeling surgical context based on video data and for automatic translation of context to MP labels. We then use our framework to create the COntext and Motion Primitive Aggregate Surgical Set (COMPASS), including six dry-lab surgical tasks from three publicly available datasets (JIGSAWS, DESK, and ROSMA), with kinematic and video data and context and MP labels. Our context labeling method achieves near-perfect agreement between consensus labels from crowd-sourcing and expert surgeons. Segmentation of tasks to MPs results in the creation of the COMPASS dataset that nearly triples the amount of data for modeling and analysis and enables the generation of separate transcripts for the left and right tools. The proposed framework results in high quality labeling of surgical data based on context and fine-grained MPs. Modeling surgical tasks with MPs enables the aggregation of different datasets and the separate analysis of left and right hands for bimanual coordination assessment. Our formal framework and aggregate dataset can support the development of explainable and multi-granularity models for improved surgical process analysis, skill assessment, error detection, and autonomy.


Interposed cartilage as a precaution against extrusions of ceramic ossicular replacement implants.
Since 1980 we have used middle ear implants of aluminum oxide ceramic in some reconstructions of the ossicular chain. We had opportunities to examine 108 ears (91 patients); the longest period of postoperative observation was 4 years. Follow-up examinations showed that the rates of extrusion and liability to extrusion can be reduced by interposing a small disk of autologous cartilage and perichondrium. Moreover, interposing cartilage trends to improve audiometric results. On the basis of our study, we strongly recommend the interposition of a disk of autologous cartilage and perichondrium between the middle ear implant and eardrum when an Al2O3 implant is used.


Systematic review and meta-analyses of the relationship between short sleep and incidence of obesity and effectiveness of sleep interventions on weight gain in preschool children.
The aim of this study is to determine (a) whether short sleep is associated with the incidence of obesity and (b) whether interventions beneficial for sleep reduce weight gain in preschool children. We systematically searched PubMed, Embase, Web of Science and Cochrane up to 12/09/2019. (a) Studies that were included were prospective, had follow-up ≥1 year, with sleep duration at baseline and required outcome measures. (b) Intervention trials with sleep intervention and measures of overweight or obesity were included. Data were extracted according to PRISMA guidelines. (a) The risk of developing overweight/obesity was greater in short sleeping children (13 studies, 42 878 participants, RR: 1.54; 95% CI, 1.33 to 1.77; p < 0.001). Sleep duration was associated with a significant change in BMI z-score (10 studies, 11 cohorts and 29 553 participants) (mean difference: -0.02 unit per hour sleep; -0.03 to -0.01; p < 0.001). (b) Four of the five intervention studies reported improved outcomes: for BMI (-0.27 kg/m2 ; -0.50 to -0.03; p = 0.03); for BMI z-score (-0.07 unit; -0.12 to -0.02; p = 0.006). Short sleep duration is a risk factor or marker of the development of obesity in preschool children. Intervention studies suggest that improved sleep may be beneficially associated with a reduced weight gain in these children.


[Structural mechanisms of the healing of heart infarctions and their correlation with changes in the peripheral blood].
Healing of myocardial infarctions and its manifestation in peripheral circulation were studied in rats, dogs, patients and autopsy cases. The importance of uncomplicated kinetics and dynamics of leucocytes in favorable outcomes of myocardial infarctions was demonstrated.


The structure of the Ni-Fe site in the isolated HoxC subunit of the hydrogen-sensing hydrogenase from Ralstonia eutropha.
The regulatory Ni-Fe hydrogenase (RH) from Ralstonia eutropha which forms a [HoxBC]2 complex functions as a hydrogen sensor under aerobic conditions. We have studied a novel Strep-tag isolate of the RH large subunit, HoxC(ST), which lacks the Fe-S clusters of HoxB, allowing for structure determination of the catalytic site by X-ray absorption spectroscopy both at the Ni and, for the first time, also at the Fe K-edge. This technique, together with Fourier-transform infrared spectroscopy, revealed a Ni-Fe site with [O1(CysS)2Ni(II)(mu-SCys)2Fe(II)(CN)2(CO)] structure in about 50% of HoxC(ST) and a [(CysS)2Fe(II)(CN)2(CO)] site lacking Ni in the remainder protein. Possibly both sites may be intermediates in the maturation process of the RH.


Gemifloxacin for community-acquired pneumonia.
Newer fluoroquinolones have become an important therapeutic choice in the treatment of community-acquired pneumonia (CAP). Gemifloxacin is one of the newest members of this class of antibiotics and has performed favourably in this indication. To analyse the microbiological activity, pharmacokinetic/pharmacodynamic properties and clinical activity of gemifloxacin in CAP, as well as the safety reported in controlled clinical studies. Literature research of English publications in the last 10 years addressing all aspects of gemifloxacin in CAP. Gemifloxacin is microbiologically the most active fluoroquinolone against Streptococcus pneumoniae--the leading pathogen of CAP. In several comparative studies gemifloxacin was highly effective and well tolerated in the treatment of mild-to-moderate severe CAP.


Veterans Administration residency programs: an overview.
At present, there are 33 Veterans Administration (VA) Medical Centers providing 1-year COE (Council on Optometric Education) accredited residency training programs for 46 recent graduates. This paper describes the types of programs available, discusses the staffing of the clinics, and details the clinical experience.


Post procedural pregnancy occurrence risk after endometrial ablation.
The objective of the study was to analyse the pregnancy rate after curettage, 1G (Endometrial resection) and 2G (Endometrial ablation) procedures in women with abnormal uterine bleeding (AUB-O,E,N) to evaluate the rate of pregnancy following these procedures and to improve pre and post-therapeutic women information. This retrospective study analyzed data extracted from the French Hospital medical information database. All hospital stays with a diagnostic code for AUB and an appropriate surgical procedure coded between 2009 and 2015 were identified. A total of 109,884 women were included. Of these, 88,165 were followed up for 18 months, 80,054 for 24 months and 33,251 for 60 months. Outcomes were compared between second generation (2G) procedures, first-generation (1G) procedures (endometrial resection) and curettage. The rate of pregnancy was the primary end point. 7863 women underwent a 2G surgical procedure (7.2%), 39,935 a 1G procedure (36.3%) and 38,923 a curettage (35.4%). The mean age of the women was 46 years (IC.95: 36.7-52.5), with no difference in age between groups. The rate of pregnancy after 2G, 1G and curettage was respectively 13 (1.5%), 617 (10.1%) and 1025 (11.1%). The primary endpoint was significantly different between 2G and 1G and curettage (p<0.0001) CONCLUSION: 2G procedures result in lower risk of pregnancy without requiring specific training for surgeons. However, endometrial ablation cannot be considered as a sterilization method nor an effective contraceptive procedure. In the absence of sterilization of either partner, women should continue to use contraception whatever their age and menstrual status.


[Intensity of negative symptoms, working memory and executive functions disturbances in schizophrenic patients in partial remission period].
The aim of the study was to assess the correlation between the level of working memory and executive functions impairment in schizophrenic subjects in their partial remission period and the intensity of psychopathological symptoms measured by PANSS scale. 45 patients with schizophrenia were included in the study (28 male and 17 female), aged 18-46 (mean 27 +/- 7) years during partial remission of psychopathological symptoms (PANSS < 70). The control group consisted in 35 age, gender and education matched healthy persons (13 male i 22 female), aged 21-49 (mean 30 +/- 8) years. To assess the intensity of psychopathological symptoms the PANSS scale was used, neuropsychological assessment included the Wisconsin Card Sorting Test (WCST), N-back test and Stroop test from the Vienna Tests Battery. In schizophrenic patients in partial remission period, the significant dysfunctions of working memory and executive functions show association with negative (not positive) schizophrenic symptoms.


Practice requirements for psychotherapeutic treatment of cancer patients in the outpatient setting-A survey among certified psychotherapists in Germany.
The aim of this study was to delineate the challenges that psychotherapists encounter when they treat cancer patients and how they organise their practices to be able to treat them. A random sample of certified psychotherapists, licensed by the health authorities, with training in psycho-oncology, was asked to complete a questionnaire covering the following issues: therapists' qualifications, organisation of the practice, dealing with appointment cancellations, financing, and networking. Practices with ≥50% cancer patients in their patient load were defined as "practices specialising in cancer" (PSC) and were compared to practices with a smaller proportion of cancer patients (non-PSC). Of 120 contacted therapists, 83 replied and 77 were eligible. The median waiting time for a first consultation was 10 days in PSC and 14 days in non-PSC (P = .05). Seventy-five of PSC and 56% of non-PSC can offer psychotherapy within 4 weeks. Time spent on dealing with the social problems of the patients was higher in PSC than in non-PSC (P = .04). They spent also more time communicating with other health care professionals such as private practice oncologists (P = .001). Cancer patients need to cancel appointments more frequently than noncancer patients (58% vs 48% cancel ≥1× per quarter). Sixty-six percent of the psychotherapists do not ask for financial reimbursement of these sessions. Psychotherapy for cancer patients in the outpatient setting requires different organisation of the practice. Sessions are cancelled more frequently, waiting time is considerably shorter, and psychotherapists communicate more often with other health care providers than in general psychotherapy.


[Assessment of microclimate at a natural gas compression plant].
The Authors evaluate microclimatic conditions in a natural gas compression plant, inside the turboblower station, and find that technical maintenance operators may be subject to heat stress hazard during summer months.


In vivo cercaria-schistosomulum transformation: inhibitory effect of oxamniquine.
High doses of oxamniquine (given intramuscularly) produced inhibition of cercaria-schistosomulum transformation following intraperitoneal injections of cercariae into mice. Cercariae, tail-less cercarial bodies, and schistosomula were recovered from the peritoneal cavity of drug-treated mice in numbers significantly different from those recovered from untreated mice. Since untreated animals induced transformation of almost all the injected cercariae, the data suggest the compound is active during the process of host-larvae adaptation.


Radiotherapy combined with immune checkpoint inhibitors in locally advanced/metastatic esophageal squamous cell carcinoma: clinical trials, efficacy and future directions.
Esophageal squamous cell carcinoma (ESCC) is a common malignancy worldwide and often diagnosed at advanced stages with poor prognosis. Combination of radiotherapy and immunotherapy seems to be a promising approach for treating ESCC. This comprehensive review article summarizes the current state of combination of radiotherapy and immunotherapy in locally advanced/metastatic ESCC, delineates the clinical trials that merit attention, and outlines unresolved issues and future research directions in this field. The clinical trial findings suggest that radio-immunotherapy combination may improve tumor response and overall survival with manageable side effects, highlighting the importance of patient selection and the necessity for further research to optimize treatment strategies. Issues such as irradiation dosage, fractionation regimen, irradiation site and technique of radiotherapy, as well as the timing, sequence and duration of combination therapy will all affect treatment outcomes, justifying further in-depth investigation.


Regulation of melanoblast and retinal pigment epithelium development by Xenopus laevis Mitf.
Mitf is a central regulator of pigment cell development that is essential for the normal development of the melanocyte and retinal pigment epithelium (RPE) lineages. To understand better the role of Mitf, we have used the Xenopus laevis experimental system to allow a rapid examination of the role of Mitf in vivo. Here, we report the function of XlMitfalpha-M on melanophore development and melanization compared with that of Slug that is expressed in neural crest cells. Overexpression of XlMitfalpha-M led to an increase in melanophores that was partly contributed by an increase in Slug-positive cells, indicating that XlMitfalpha-M is a key regulator of melanocyte/melanophore development and melanization. Moreover, overexpression of a dominant-negative form of XlMitfalpha led to a decrease in the number of melanophores and induced abnormal melanoblast migration. We also observed an induction of ectopic RPE and extended RPE by overexpression of XlMitfalpha-M and possible interactions between XlMitfalpha and several eye-related genes essential for normal eye development.


A case of acute basophilic leukemia arising from chronic myelogenous leukemia with development of t(7;8)(q32;q13).
Acute basophilic leukemia (ABL) is an uncommon form of acute myelogenous leukemia recently recognized as a distinct entity in the World Health Organization classification of myeloid malignancies. A case is presented of ABL arising from chronic myelogenous leukemia with development of t(7;8)(q32;q13). Discussion includes a literature review.


Pathology of the cardiac conduction system in myotonic dystrophy: a study of 12 cases.
In 12 autopsy cases of myotonic dystrophy, the most frequently observed histopathologic lesions of the cardiac conduction system were fibrosis, fatty infiltration and atrophy. Fibrosis involved the sinus node in 6 cases, atrioventricular (AV) node in 7, AV bundle in 8, bundle branches in 10 and ventricular myocardium in 11. Fatty infiltration was observed in the sinus node in two cases, AV node in two, AV bundle in six, bundle branches in one and ventricular myocardium in nine. Atrophy was prominent in the AV bundle in five and bundle branches in eight. Lymphocytes infiltrated the conduction system in three cases and were associated with myotonic dystrophy in two and varicella myocarditis in one. Ventricular myocytes were hypertrophied in seven cases, vacuolated in three and exhibited disarray in two. The distribution and extent of conduction system lesions tended to correspond to antemortem electrocardiographic abnormalities, including prolonged PR interval in six cases, intraventricular conduction delay in six and bundle branch block in four. Cardiac involvement by myotonic dystrophy may have contributed to sudden death in four cases.


Isolation and HIV-1 infection of primary human microglia from fetal and adult tissue.
Glial inflammation, principally involving astrocytes and microglia, underlies the pathogenesis of a broad range of neurodegenerative disorders, including, most notably, human immunodeficiency virus (HIV-1)-associated dementia. Indeed, for the latter, disease mechanisms are attributed to viral infection and activation of microglia and perivascular macrophages and their resultant neurotoxic activities. Although monocyte-derived macrophages have served as models for microglia, they are limited both qualitatively and quantitatively in their immune responses and susceptibility to viral infection. Thus, the acquisition of primary human microglial cells is critical for laboratory studies of human neurological disease. In this chapter, we provide detailed methods of isolation, cultivation, characterization, HIV-1 infection, and experimental applications of primary human fetal and adult microglial cells, with particular emphasis on studies of HIV-1 neuropathogenesis.


Study of the counts, species and characteristics of the yeast population during the manufacture of dry-cured "lacón". Effect of salt level.
The aim of this work was to study the yeast population during the manufacture of dry-cured "lacón" (a Spanish traditional meat product) and the effect of the salting time. For this study, six batches of "lacón" were manufactured with three different salting times (LS (3 days of salting), MS (4 days of salting) and HS (5 days of salting)). Yeast counts increased significantly (P < 0.001) during the whole process from 2.60 to 6.37 log cfu/g. An increased length of salting time did not affect yeast counts throughout the manufacture of dry-cured "lacón", although the highest yeast counts were obtained from LS batches. A total of 226 isolates were obtained from dry-cured "lacón" during drying-ripening stage, of which 151 were yeasts and were identified at the species level using molecular techniques. The total of 151 identified yeasts belonged to 4 different genera: Debaryomyces, Candida, Cryptococcus and Rhodotorula. Debaryomyces hansenii was the most abundant species isolated throughout the whole process as much in the interior as in the exterior of the pieces of three salt levels of "lacón" studied, while Candida zeylanoides was only isolated from the interior of MS and HS batches and from the exterior of LS and HS groups, but at lesser proportion than D. hansenii.


Antigenic relationships between rotaviruses from different species as studied by neutralization and immunofluorescence.
Antigenic relationships between the Wa, Hochi, Ito, Hosokawa and Nemoto strains of human rotavirus, the SA-11 strain of simian rotavirus, the Lincoln strain of bovine rotavirus, the OSU strain of porcine rotavirus and the R-2 strain of lapine rotavirus, which were all established in cell culture, were studied by neutralization (NT) and immunofluorescence (IF) using guinea pig antisera against these strains. The strains could not be distinguished by indirect IF staining of infected cells because of marked cross reactions. On the other hand, the NT test clearly distinguished these strains, although there was a one-way antigenic relationship between some of them. The five strains of human rotavirus were clearly distinguished from each other by NT test. These human strains exhibited much higher homologous titers than heterologous titers, the former being at least eight-fold higher than the latter. The presence of at least five serotypes of human rotavirus was indicated.


Generation of an Abcc8 heterozygous mutation human embryonic stem cell line using CRISPR/Cas9.
The gene of ATP-binding cassette subfamily C member 8 (Abcc8) is cytogenetically located at 11p15.1 and encodes the sulfonylurea receptor (SUR1). SUR1 is a subunit of ATP-sensitive potassium channel (KAPT) in the β-cell regulating insulin secretion. Mutations of ABCC8 are responsible for congenital hyperinsulinism (CHI). Here we reported that an Abcc8 heterozygous mutant cell line was generated by CRISPR/Cas9 technique with 1bp insertion resulting in abnormal splicing on human embryonic stem cell line H1. The phenotypic characteristics of this cell line reveal defective KATP channel and diazoxide-responsive that provides ideal model for molecular pathology research and drug screening for CHI.


Highly potent and stereoselective effects of the benzoic acid derivative AZ-DF 265 on pancreatic beta-cells.
1. Mouse islets were used to define the characteristics and study the mechanisms of the stimulation of insulin release by compound AZ-DF 265, 4-[[N-(alpha-phenyl-2-piperidino-benzyl) carbamoyl]methyl] benzoic acid, a substituted benzoic acid with an asymmetric carbon atom. 2. At a non-stimulatory concentration of glucose (3 mM), (-)-AZ-DF 265 reversibly inhibited 86Rb efflux from islet cells, depolarized the beta-cell membrane, induced electrical activity, stimulated 45Ca efflux, and triggered insulin release. Maximum inhibition of 86Rb efflux occurred at 0.03 microM (-)-AZ-DF 265, whereas the threshold concentration for stimulation of release was 0.1 microM. Omission of extracellular Ca2+ abolished all effects of the drug but the inhibition of 86Rb efflux. 3. At a stimulatory concentration of glucose (10 mM), (-)-AZ-DF 265 reversibly increased 86Rb efflux, potentiated electrical activity, augmented 45Ca efflux, and increased insulin release. Maximum stimulation of 86Rb efflux and insulin release was obtained with 0.03 microM (-)-AZ-DF 265. Omission of extracellular Ca2+ abolished all effects of the drug. 4. The potency of (-)-AZ-DF 265 was similar to that of glibenclamide, whereas the (+)-enantiomer was about 10 times less potent on 86Rb efflux and insulin release. 5. It is concluded that, like sulphonylureas, compound AZ-DF 265 decreases K+ permeability of the beta-cell membrane and thereby causes depolarization. This activates voltage-dependent Ca channels, permits Ca2+ influx and eventually stimulates insulin release. Its stereoselectivity may help to elucidate the mechanisms of K channel blockade and, hence, lead to the design of more potent and specific insulinotropic drugs.


Expression analyses of stress-responsive genes in the hermatypic coral Acropora tenuis and its symbiotic dinoflagellates after exposure to the herbicide Diuron.
Diuron is one of the most frequently applied herbicides in sugarcane farming in southern Japan, and Australia. In addition, it is used as a booster substance in copper-based antifouling paints. Due to these various uses, Diuron is released into the marine environment; however, little information is available on gene expression in corals and their symbiotic algae exposed to Diuron. We investigated the effects of Diuron on stress-responsive gene expression in the hermatypic coral Acropora tenuis and its symbiotic dinoflagellates. After seven days of exposure to 1 µg/L and 10 µg/L Diuron, no significant changes in the body colour of corals were observed. However, quantitative reverse transcription-polymerase chain reaction analyses revealed that the expression levels of stress-responsive genes, such as heat shock protein 90 (HSP90), HSP70, and calreticulin (CALR), were significantly downregulated in corals exposed to 10 µg/L of Diuron for seven days. Moreover, aquaglyceroporin was significantly downregulated in corals exposed to environmentally relevant concentrations of 1 µg/L Diuron. In contrast, no such effects were observed on the expression levels of other stress-responsive genes, such as oxidative stress-responsive proteins, methionine adenosyltransferase, and green/red fluorescent proteins. Diuron exposure had no significant effect on the expression levels of HSP90, HSP70, or HSP40 in the symbiotic dinoflagellates. These results suggest that stress-responsive genes, such as HSPs, respond differently to Diuron in corals and their symbiotic dinoflagellates and that A. tenuis HSPs and CALRs may be useful molecular biomarkers for predicting stress responses induced by the herbicide Diuron. The online version contains supplementary material available at 10.1007/s42995-023-00183-0.


PADI-web 3.0: A new framework for extracting and disseminating fine-grained information from the news for animal disease surveillance.
PADI-web (Platform for Automated extraction of animal Disease Information from the web) is a biosurveillance system dedicated to monitoring online news sources for the detection of emerging animal infectious diseases. PADI-web has collected more than 380,000 news articles since 2016. Compared to other existing biosurveillance tools, PADI-web focuses specifically on animal health and has a fully automated pipeline based on machine-learning methods. This paper presents the new functionalities of PADI-web based on the integration of: (i) a new fine-grained classification system, (ii) automatic methods to extract terms and named entities with text-mining approaches, (iii) semantic resources for indexing keywords and (iv) a notification system for end-users. Compared to other biosurveillance tools, PADI-web, which is integrated in the French Platform for Animal Health Surveillance (ESA Platform), offers strong coverage of the animal sector, a multilingual approach, an automated information extraction module and a notification tool configurable according to end-user needs.


De novo biosynthesis of polyunsaturated fatty acids in the cockroach Periplaneta americana.
The de novo biosynthesis of 6,9,12-linolenic acid, 11,14-eicosadienoic acid, 5,11,14-eicosatrienoic acid, and arachidonic acid was demonstrated in adult female cockroaches, Periplaneta americana. These four polyunsaturated fatty acids (PUFA) were present primarily in the phospholipid (PL) fraction of both males and females. They were purified by AgNO3 thin-layer chromatography and high pressure liquid chromatography. The double bond positions of the major isomer of eicosatrienoic acid were shown to be at the delta 5,11,14 positions by gas chromatography-mass spectrometry (GC-MS) of both methoxy and epoxide derivatives and gas-liquid chromatography (GLC) and GC-MS of ozonolysis products. The other PUFAs cochromatographed with standards on both packed and capillary GLC columns. The in vivo incorporation of [1-14C]acetate into 5,11,14-eicosatrienoic acid, 11,14-eicosadienoic acid, 6,9,12-linolenic acid, and arachidonic acid was demonstrated by radio-GLC and radio-HPLC and for 5,11,14-eicosatrienoic acid by radio-GLC of ozonolysis products. The latter technique clearly demonstrated that the entire eicosatrienoic acid molecule was labeled. Thoracic tissue contained the highest amount of radiolabeled 5,11,14-eicosatrienoic acid (1.6% of total radioactivity incorporated into PL) while radiolabeled 11,14-eicosadienoic acid was found primarily in abdominal epidermal tissue (2% of total radioactivity incorporated into PL). Radiolabeled arachidonic and 6,9,12-linolenic acids comprised 0.1 and 0.02%, respectively, of the total radioactivity in the PL fraction. These data document the de novo biosynthesis of di-, tri-, and tetraunsaturated fatty acids in the American cockroach, and indicate that this animal can desaturate on both sides of the delta 9 double bond of oleic acid.


Acyclic Isoprenoid Attenuates Lipid Anomalies and Inflammatory Changes in Hypercholesterolemic Rats.
The present study was aimed to explore the antihyperlipidemic and anti-inflammatory effect of acyclic isoprenoid on high fat diet fed rats. Hypercholesterolemia was induced by the diet comprising of the normal rat chow 84.3%, lard 5%, yolk powder 10%, cholesterol 0.2% and 0.5% bile salt were fed to the rats for the period of 8 weeks. The results showed that abnormally elevated levels of plasma lipid profiles. Three different doses of acyclic isoprenoid (20, 40 and 80 mg/kg b.w/day) were administered orally to hypercholesterolemia suffering rats for the period of 30 days. Among these three doses of acyclic isoprenoid, the dose 80 mg/kg b.w. was significantly decreased the plasma lipid profiles when compared to other two doses. The effect produced by acyclic isoprenoid (80 mg/kg b.w) was comparable to that of simvastatin. Therefore, 80 mg/kg b.w was fixed as a effective dose and used for further analyses. Acyclic isoprenoid administration reinstated the elevated levels of cardiac and inflammatory markers in both blood and serum of hypercholesterolemic rats. In addition, acyclic isoprenoid administration decreased activity of 3-hydroxy 3-methyl-glutaryl-CoA reductase and increased the activity of lecithin cholesterol acyl transferase. These findings suggest that the administration of acyclic isoprenoid was potentially ameliorated the cardiac marker enzymes and inflammatory markers in addition to its antihypercholesterolemic effect.


Intranuclear inclusions in pericytes of the hypothalamus of the rat.
This paper deals with the ultrastructure of two types of intranuclear inclusions, "nuclear bodies" and "membranous lamellar bodies", present in hypothalamic pericytes of intact adult rats. The nuclear bodies exhibited "simple" and "granular" forms, whereas the membranous lamellar bodies were entirely made up of myelin-like membrane whorls. The occurrence of these bodies in nuclei of pericytes has never been previously reported. The origin and functional meaning of such structures is discussed in the light of recent ultrastructural and biochemical studies on nuclear inclusions.


Theta dominates cross-frequency coupling in hippocampal-medial entorhinal circuit during awake-behavior in rats.
Hippocampal theta and gamma rhythms are hypothesized to play a role in the physiology of higher cognition. Prior research has reported that an offset in theta cycles between the entorhinal cortex, CA3, and CA1 regions promotes independence of population activity across the hippocampus. In line with this idea, it has recently been observed that CA1 pyramidal cells can establish and maintain coordinated place cell activity intrinsically, with minimal reliance on afferent input. Counter to these observations is the contemporary hypothesis that CA1 neuron activity is driven by a gamma oscillation arising from the medial entorhinal cortex (MEC) that relays information by providing precisely timed synchrony between MEC and CA1. Reinvestigating this in rats during appetitive track running, we found that theta is the dominant frequency of cross-frequency coupling between the MEC and hippocampus, with hippocampal gamma largely independent of entorhinal gamma.


Lipid peroxidation-induced inhibition of gamma-aminobutyric acid uptake in rat brain synaptosomes: protection by glucocorticoids.
Incubation of rat brain synaptosomes with xanthine and xanthine oxidase (X/XO) resulted in an inhibition of gamma-aminobutyric acid (GABA) uptake. The inhibitory effects of X/XO were temperature- and time-dependent, and were characterized by an increased Km for GABA and a decreased Vmax. Inhibition of GABA uptake by X/XO was associated with both the formation of malonyldialdehyde (MDA) and conjugated dienes, indicating that lipid peroxidation was involved. Studies with catalase, superoxide dismutase (SOD), mannitol, and chelated iron suggested that hydroxyl radical (OH X) was probably responsible for the initiation of lipid peroxidation. Both the peroxidation of synaptosomal membranes and the inhibition of GABA uptake by X/XO were enhanced by the addition of ADP and FeCl2. The X/XO-induced inhibition of GABA uptake by synaptosomes could be prevented by preincubation of synaptosomes with certain glucocorticoids prior to X/XO exposure. Methylprednisolone sodium succinate (MPSS), dexamethasone sodium phosphate (DMSP), and prednisolone sodium succinate (PSS) all prevented the inhibition of GABA uptake by X/XO. MPSS was most effective at concentrations around 100 microM, DMSP was slightly more potent, and PSS was optimal at around 300 microM. On the other hand, hydrocortisone sodium succinate (HCSS) was ineffective at preventing X/XO-induced inhibition of GABA uptake at concentrations up to 3 mM. The steroids are presumed to work through a mechanism that blocked the formation of lipid peroxides, as MPSS inhibited the formation of conjugated dienes in synaptosomes exposed to X/XO at a concentration that also protected GABA uptake.


Functional mimicry of the LDL receptor-associated protein through multimerization of a minimized third domain.
The LDL receptor-associated protein (RAP) is a ligand for the LDL receptor-related protein (LRP1). The first and third domains of RAP can each bind to one of many sequence-related pairs of complement-type repeats (CR) found within the LRP1 ectodomain. Multiple sites of interaction between the multivalent RAP ligand and the multivalent LRP1 receptor yield strong binding avidity for the complex. The third domain of RAP can be significantly truncated, with material retention of monovalent CR pair-binding affinity, provided that the minimized sequence is stabilized with an intramolecular disulfide bond. We demonstrate that the avidity of full-length RAP for LRP1 in vitro can be partially reconstituted by assembly of truncated, disulfide-linked RAP peptides on tetravalent streptavidin or bivalent immunoglobulin scaffolds. The peptide complex with streptavidin shows pronounced hepatotropism in vivo, replicating the biodistribution of full-length RAP.


Fungal adaptation to the mammalian host: it is a new world, after all.
Adaptation to environmental conditions is key to fungal survival during infection of human hosts. Although the host immune system is often considered the primary obstacle to fungal colonization, invading fungi must also contend with extreme abiotic stresses. Recent work with human pathogenic fungi has uncovered systems for detecting and responding to changes in temperature, carbon source, metal ion availability, pH, and gas tension. These systems play a major role in adaptation to the host niche and are essential factors for persistence in a mammalian host. Future investigations into fungal responses to these and other abiotic components of the host environment have the potential to uncover novel targets for anti-fungal therapy.


Genetic variation in the proto-oncogene SKI and risk for orofacial clefting.
SKI is a proto-oncogene that is required for development of the central nervous system and skeletal muscle, and is involved in specifying selected cranial neural-crest-derived craniofacial structures. To identify genetic variants within the SKI gene and investigate the potential association between SKI polymorphisms and risk for orofacial defects, we initially re-sequenced the gene. DNA re-sequencing of all seven exons of the SKI gene was performed on 100 control samples. Subsequently, we genotyped 394 samples (148 CLP cases, 99 CP cases, and 147 control infants) for a novel SNP identified in the DNA re-sequencing effort using restriction fragment length polymorphism (RFLP) analysis. We identified one polymorphism in exon 1 of the SKI gene (257C>G) from controls. This SNP resulted in an amino acid change from alanine to glycine (A62G, GenBank Accession No. NM_003036). Among all samples genotyped by the RFLP method, variants (CG, GG) were found in 10.5% of the cases, compared to a prevalence of 17.7% in the controls. The odds ratio was calculated to be 0.6, with a 95% confidence interval (CI) of 0.3-1.0. In a population of California infants with craniofacial defects, a novel polymorphism of the SKI gene was found to be associated with a decreased risk for orofacial defects. The function of this polymorphism and how it might confer protection to the embryo against craniofacial malformations is currently under investigation in our laboratory.


What Donation Features are Related to Complications in Donation after Circulatory Death Kidney Transplant? Analysis of Risk Factors for Complication after Circulatory Death Kidney Transplant.
Complications in donation after circulatory death (DCD) kidney transplants (KT) are barely described, while in some urological complications the cause is unknown. The aim of this study is to describe surgical and urological complications and analyze what donation features could be involved. A prospective, single center study was performed from 2016 to 2019 including all KT from controlled cardiac death donors (cDCD). A total of 86 cDCD KT were included in the study. Recipient BMI, residual urine output (RUO) <500 mL/day, delayed graft function (DGF), and wound complication were related to UTI (p = 0.020, p = 0.008, p = 0.016, and p = 0.004, respectively). Features related to early graft nephrectomy were recipient BMI and recipients with diabetes mellitus (DM) (p = 0.025 and p = 0.036, respectively). DM in recipients was significantly associated with hematuria (p = 0.046). Urinary leak (UL) was associated to vascular complication and ureteral stricture (US) (p = 0.029 both). UL and lymphocele were associated to US (p = 0.029 both). Features related to lymphocele were recipient BMI and US (p = 0.028 and p = 0.029, respectively). History of previous transplant, time from cardiac arrest (CA) to cold flush, and DGF, were associated to wound complication (p = 0.040, p = 0.011 and p = 0.016, respectively). Surgical and urological complications after KT are an important issue to resolve. Our data revealed an association between RUO <500 mL/day, DGF, and wound complication with urinary infection, as well as between recipient DM and hematuria. Recipient BMI and DM were related to early graft nephrectomy. Vascular complications were associated with urinary leak, and lymphocele with US. Finally, wound complication was related to previous transplant, DGF, and time from CA to cold flush. This data revealed interesting associations between donor and recipient features and cDCD KT complications, providing more information to improve prevention and management.


Advanced multifunctional nano-lipid carrier loaded gel for targeted delivery of 5-flurouracil and cannabidiol against non-melanoma skin cancer.
Non-melanoma skin cancer is one of the most common malignancies reported around the globe. Current treatment therapies fail to meet the desired therapeutic efficacy due to high degree of drug resistance. Thus, there is prominent demand in advancing the current conventional therapy to achieve desired therapeutic efficacy. To break the bottleneck, nanoparticles have been used as next generation vehicles that facilitate the efficient interaction with the cancer cells. Here, we developed combined therapy of 5-fluorouracil (5-FU) and cannabidiol (CBD)-loaded nanostructured lipid carrier gel (FU-CBD-NLCs gel). The current investigation has been designed to evaluate the safety and efficacy of developed 5-Flurouracil and cannabidiol loaded combinatorial lipid-based nanocarrier (FU-CBD NLCs) gel for the effective treatment of skin cancer. Initially, confocal microscopy study results showed excellent uptake and deposition at epidermal and the dermal layer. Irritation studies performed by IR camera and HET cam shows FU-CBD NLCs was much more tolerated and less irritant compared to conventional treatment. Furthermore, gamma scintigraphy evaluation shows the skin retention behavior of the formulation. Later, in-ovo tumor remission studies were performed, and it was found that prepared FU-CBD NLCs was able to reduce tumor volume significantly compared to conventional formulation. Thus, obtained results disclosed that permeation and disposition of 5-FU and CBD into different layers of the skin FU-CBD NLCs gel could be more potential carrier than conventional gel. Furthermore, prepared formulation showed greater tumor remission, better survival rate, reduction in tumor number, area, and volume with improved biochemical profile. Thus, prepared gel could serve as a promising formulation approach for the skin cancer treatment.


Translational level of acetylcholine receptor α mRNA in mouse skeletal muscle is regulated by YB-1 in response to neural activity.
Y-box-binding protein 1 (YB-1) binds to mRNAs and affects translation. In this study, we focused on skeletal muscle, in which YB-1 expression is restricted to the early postnatal period, and found that YB-1 binds to acetylcholine receptor α-subunit (AChR α) mRNA. Although transcription of the AChR α gene is known to be regulated by myogenic transcription factors, translational control of the mRNA in response to neuromuscular transmission has not been examined. In undifferentiated C2C12 myoblasts, expression of AChR α remained at a low level. However, translation of the mRNA was increased by knockdown of YB-1. Continued overexpression of YB-1 prevented the cells from differentiating. In myotubes, which show clustering of AChRs, translation of the mRNA was induced within 3h after treatment with nicotine. The effect of nicotine was inhibited by α-bungarotoxin, and in the presence of cycloheximide the level of AChR α was reduced, even after nicotine treatment. Sucrose gradient analysis revealed that in nicotine-treated myotubes, YB-1-containing polysomes were shifted to the heavier-sedimenting fractions, and showed an apparent decrease in the amount of YB-1 bound to AChR α mRNA. These results suggest that in skeletal muscle cells, neural activity reduces the molar ratio of YB-1 relative to its binding AChR α mRNA, leading to an increase of ribosome binding to the mRNA, and thus activating translation. Furthermore, in postnatal growing mice, as has already been shown, the level of AChR α mRNA declined during the early period with maturation of neuromuscular synapses, but the translation level was found to increase transiently at postnatal day 10, when the level of YB-1 was markedly reduced. It is suggested that although the level of AChR α mRNA is reduced, the translation can be induced by alteration of the ratio of YB-1 protein to the mRNA.


Biological characterisation of a recombinant Atlantic salmon type I interferon synthesized in Escherichia coli.
Type I (alpha/beta) interferons (IFNs) are a family of cytokines that stimulate the expression of numerous proteins that mediate an antiviral state in uninfected cells. Two Atlantic salmon (Salmo salar) IFN-alpha (SasaIFN-alpha1 & 2) genes have previously been cloned and both were found to contain a putative N-linked glycosylation site. Recombinant SasaIFN-alpha1 (rSasaIFN-alpha1) produced in eukaryotic systems has repeatedly been shown to confer antiviral properties. However, different IFN-alpha subtypes may exhibit differential antiviral activities and be subject to glycosylation. To evaluate the potential therapeutic impact of a rSasaIFN-alpha, the mature form of the SasaIFN-alpha2 protein was produced in a high-level Escherichia coli expression system. Expression of the rSasaIFN-alpha2 was detected by SDS-PAGE and Western blot, and its identity was confirmed by mass spectrometry. In the homologous Chinook salmon embryonic (CHSE-214) cell line, the rSasaIFN-alpha2 incited early expression of the IFN-induced Mx protein and exhibited high antiviral activity of about 2.8 x 10(6) U mg(-1) against infectious pancreatic necrosis virus (IPNV). Conversely, antiviral protection by rSasaIFN-alpha2 was not observed in a heterologous Japanese flounder embryo (HINAE) cell line. Hence, a biologically active form of rSasaIFN-alpha2 was successfully produced using a glycosylation-deficient prokaryotic system and purified to homogeneity, suggesting that glycosylation is not required for its antiviral activity.


The role of coagulation on the fate of PFAS, brominated flame retardants and other trace contaminants in tertiary wastewater treatment for phosphorus control.
Coagulant dosing to achieve low phosphorus concentrations in wastewater effluents may favour the removal of trace organics such as pharmaceuticals, plasticisers and flame retardants. Nevertheless, the behaviour of trace organics in coagulation processes is currently poorly understood because of the complex interactions between these compounds, the coagulants and dissolved organic matter (DOM). This study assessed the coagulation removal from synthetic secondary effluent of twenty-four compounds including ten PFAS and four brominated flame retardants. Testing involved two coagulants (alum, ferric chloride) and five DOM surrogates (resorcinol, benzoic acid, citric acid, tannic acid, humic acid); DOM surrogates had assorted molecular weights, structures, charges, and hydrophobicity. With coagulant doses of 14 mg Fe/L and 4 mg Al/L, ten trace organics were removed by >30 % in the presence of at least one DOM surrogate. Humic acid effected the highest removals owing to complexation of trace organics and subsequent co-removal by adsorption or sweep floc. For instance, removal extents for three brominated diphenyl ethers were 60 to 75 % with Al and 50 to 88 % with Fe (initial concentration 0.4 to 0.8 ng/L); PFTDA, a long-chain PFAS, was removed by 87 and 91 % with Fe in the presence of tannic or humic acid, respectively (initial concentration 0.03 μg/L). The varying coagulation performance of different treatment works in terms of trace substance removal can be explained because of the site-specific DOM characteristics. Addition of humic acids as complexing agents has the potential to improve the removal of hydrophobic trace substances, including some long-chain PFAS and brominated flame retardants.


The impact of cation concentration on Microcystis (cyanobacteria) scum formation.
Cyanobacterial scums at the surface of the lakes are potentially harmful phenomena with increasing occurrence in the last decades, and the causes that lead to their formation are still an unresolved issue. In order to better understand what triggers the scums, we investigated the effect of several Mg2+ and Ca2+ ion concentrations in promoting them in eight Microcystis aeruginosa strains. The possibility to prevent scum formation by using the ion chelator EDTA was also explored. We found that in some strains the cell aggregation takes place under lower ion source concentrations (20 mM MgSO4 or CaCl2), while in others this phenomenon does not occur even at 60 mM concentration. The scum formation correlated to the amount of extracellular polymeric substances (between 234 and 351 µg/cell). EDTA failed to prevent the scum formation in most strains, and in turn it caused cell lysis followed by the release of cellular content into the culture medium. We emphasize the relevance of these results for cyanobacterial scum formation in the environment and we also suggest that controlling the salinity of the medium (by manipulating the ion concentration) is a potentially efficient method for biomass harvesting in large ponds/tanks.


On the calculation of sugar concentration in flower nectar.
There are several sources of potential error in calculating the concentration or energy value of floral nectar. Errors resulting from confusing data become substantial with increasing concentration. The different methods of expressing sugar concentration are here clarified, and the correct methods of converting from one to the other are provided. Refractometers in use in field studies usually read on a weight per total weight basis; this is recommended as the mode of statement. The perils of oversimplifying conversions from this mode, as is often done, are pointed out.


Serial transmission of spongiocytic hepatitis to woodchucks (possible association with a specific delta strain).
Outbreaks of severe hepatitis have been reported from Africa and South America. Description of the cases has shown the histological hallmark to be the presence of ballooning hepatocytes with fat drops surrounding the nucleus (spongiocytes or morula cells). Experimental reproduction of this syndrome for the verification of a possible role of a specific HDV strain was performed by the inoculation of serum and liver extracts from African patients (Bangui-Central African Republic), who died with this syndrome, into American woodchuck carriers of WHV (WC 231,144), the results of which were then compared with animals inoculated with a reference wild HDV strain (WC 300,173,154), and those which received material from a European fulminant HDV case (WC 88,93). Following the initial inoculation, the animals receiving African inocula had a delayed anti-HDV seroconversion, high mortality and showed the presence of spongiocytes, while the other animals had a classical evolution of HDV superinfection in woodchucks. Furthermore, the African inocula caused less inhibition of WHV replication, as well as a predominant cytoplasmic expression of HDAg, in contrast to the animals which received the other inocula. The second passage experiments gave similar results. We conclude that this peculiar form of HDV fulminant hepatitis can be experimentally reproduced and might be specifically related to a more pathogenic strain.


Enhanced synergetic antibacterial activity by a reduce graphene oxide/Ag nanocomposite through the photothermal effect.
Multidrug-resistant (MDR) bacterial strains have led to notable heathy threats to human beings. The demand for the development of effective antibacterial materials is increasing. Silver nanoparticles (AgNPs) and graphene-based nanomaterials are two major types of nanomaterials that are studied to inhibit and/or kill bacteria. In this study, by combining the excellent photothermal effect of graphene and antibacterial activity of AgNPs, we have applied reduced graphene oxide/silver (RGO/Ag) nanocomposite to destroy the MDR bacteria. The antibacterial activity of the RGO/Ag nanocomposite was systematically investigated using a regular bacterium of Escherichia coli (E. coli) and an MDR bacterium of Klebsiella pneumoniae (Kp). Compared with AgNPs, graphene oxide (GO) and RGO, the RGO/Ag nanocomposite showed significant higher antibacterial efficiency for both regular bacteria and MDR bacteria. Under a near-infrared (NIR) irradiation (0.30 W/cm2 for 10 min), the RGO/Ag nanocomposite demonstrated an enhanced synergetic antibacterial activity through the photothermal effect. Nearly 100 % of E. coli and Kp were killed by the treatment of 15 μg/mL and 30 μg/mL of RGO/Ag nanocomposite, respectively. Moreover, a membrane integrity assay and a reactive oxygen species (ROS) assay demonstrated that the RGO/Ag nanocomposite under NIR irradiation induced the cell membrane disruption and generation of ROS, providing possible mechanisms for their high antibacterial activity besides the photothermal effect. Finally, the cytotoxicity of the RGO/Ag nanocomposites toward different mammalian cells was studied, the cell viabilities retained above 60 % at higher concentrations of RGO/Ag, indicating that the RGO/Ag nanocomposites may be a low cytotoxic, efficient antibacterial agent with the irradiation.


The effect of kidney transplantation on distal tubular vacuolar H+-ATPase.
The presence of vacuolar (v)H+-ATPase in distal tubule alpha-intercalated cells is essential for hydrogen excretion and maintenance of acid-base homeostasis. Loss of vH+-ATPase after kidney transplantation could cause posttransplant distal renal tubular acidosis (dRTA). Immunostaining of the kidney specific vH+-ATPase of cortical collecting duct cells (CCT) was performed in 37 kidney biopsies taken immediately prior to transplantation and after engraftment (median [range]: 10 [1-181] months). Apical or intracytoplasmatic staining intensity was classified as grade 0 (absent), grade 1 (weak), or grade 2 (strong), and positive cells expressed as percentage of all CCT cells. In addition, kidney biopsies were scored for damage by the Banff schema. Serum and urinary pH, anion gap, and serum potassium were obtained for the diagnoses of dRTA. Fourteen transplant recipients had dRTA type I, 5 had rate-limited RTA, six had type IV dRTA, and 12 had no RTA. In pretransplant biopsies, 40% [3-77%] of CCT cells were positive for vH+-ATPase but only 17% [0-39%] after transplantation (P<0.0001). The loss of vH+-ATPase expression was similar in patients with dRTA type I (-21%), type IV (-25%), rate limited RTA (-21%), or no RTA (-29%). The decrease affected predominantly the apical proton pump expression. The individual loss of vH+-ATPase expression was not related to the time elapsed since transplantation, immunosuppressive drugs, acute transplant rejection, or tubulointerstitial changes. Kidney transplantation leads to a general decrease of distal tubular vH+-ATPase expression. Loss of proton pump activity occurs unrelated to immunosuppressive therapy or transplant related histologic changes.


Synergistic effect of eugenol, carvacrol, thymol, p-cymene and γ-terpinene on inhibition of drug resistance and biofilm formation of oral bacteria.
Dental caries remains the most prevalent oral infectious disease worldwide. In this study, the antibacterial and the antibiofilm activities of five essential oils (EO's): eugenol (EUG), carvacrol (CAR), thymol (TYH), p-cymene (CYM) and γ-terpinene (TER) were tested (alone or in combinaison with tetracycline) against oral bacteria. In addition, their potential roles to enhance the accumulation of ethidium bromide (EtBr) in bacterial cells were tested. Our results indicated that EO's induced a selective antimicrobial activity. A synergistic effect of EO's and tetracycline (TET) was noticed with a reduction rate ranged from 2 to 8-fold. In addition, the efflux of EtBr was inhibited with a decrease in loss of EtBr from the bacteria. On the other hand a significant anti-biofilm activities of EO's (alone or combined with antibiotics) was noticed. In conclusion the tested EO's may be considered as a potential natural source with a resistance-modifying activity and may be applied to eradicate bacterial biofilm.


Glycemic Management in the Operating Room: Screening, Monitoring, Oral Hypoglycemics, and Insulin Therapy.
This review provides a literature update and practical outline for the management of diabetes and stress hyperglycemia for adult surgical patients in the pre- and intraoperative settings. Hyperglycemia in surgical patients has been associated with increased risk of complication in both diabetic and non-diabetic patients in the perioperative setting. While current recommended perioperative blood glucose target is < 180 mg/dL (10 mmol/L), optimal outcomes may require different treatment targets for diabetic versus non-diabetic patients. Hemoglobin A1C level is associated with elevated risk of hyperglycemia and adverse outcomes, but there is insufficient evidence to recommend routine preoperative testing or optimal values in elective surgical patients. Day of surgery blood glucose testing and treatment are recommended in the perioperative period, and anesthetic management includes appropriate patient selection for use of subcutaneous insulin, intravenous insulin infusions, and insulin pumps. Additionally, administration of both intravenous and perineural dexamethasone is associated with increased blood glucose levels and clinicians should consider the risk benefit ratio in surgical patients. For enhanced recovery after surgery protocols, further evidence is needed to support routine use of carbohydrate loading in diabetic patients. Optimal perioperative care includes screening at-risk patients, use of preoperative oral hypoglycemics and home insulin, anesthetic type and medication selection, blood glucose testing, and treatment for hyperglycemia in the operating room. Partnerships with surgery and endocrinology teams aid optimal postoperative management and discharge planning.


Acyl-coenzyme A:cholesterol acyltransferase 1 blockage enhances autophagy in the neurons of triple transgenic Alzheimer's disease mouse and reduces human P301L-tau content at the presymptomatic stage.
Patients with Alzheimer's disease (AD) display amyloidopathy and tauopathy. In mouse models of AD, pharmacological inhibition using small molecule enzyme inhibitors or genetic inactivation of acyl-coenzyme A (Acyl-CoA):cholesterol acyltransferase 1 (ACAT1) diminished amyloidopathy and restored cognitive deficits. In microglia, ACAT1 blockage increases autophagosome formation and stimulates amyloid β peptide1-42 degradation. Here, we hypothesize that in neurons ACAT1 blockage augments autophagy and increases autophagy-mediated degradation of P301L-tau protein. We tested this possibility in murine neuroblastoma cells ectopically expressing human tau and in primary neurons isolated from triple transgenic AD mice that express mutant forms of amyloid precursor protein, presenilin-1, and human tau. The results show that ACAT1 blockage increases autophagosome formation and decreases P301L-tau protein content without affecting endogenous mouse tau protein content. In vivo, lacking Acat1 decreases P301L-tau protein content in the brains of young triple transgenic AD mice but not in those of old mice, where extensive hyperphosphorylations and aggregation of P301L-tau take place. These results suggest that, in addition to ameliorating amyloidopathy in both young and old AD mice, ACAT1 blockage may benefit AD by reducing tauopathy at early stage.


The influence of latitude on the concentration of vitamin D3 and 25-hydroxy-vitamin D3 in Australian red meat.
There is little information on the vitamin D content of Australian red meat or on the possible influence of latitude on this content. To determine the content of vitamin D3 and 25-hydroxy-vitamin D3 (25OHD3), lamb and beef were analysed from 34° S with LC-IT-MS. To investigate the possible influence of latitude on vitamin D in meat, the lean meat and fat from five cuts of beef were analysed from 17° S and 41° S. Lamb contained 0.10μg vitamin D3/100g and 0.20μg 25OHD3/100g lean meat, while beef contained 0.12μg vitamin D3 and 0.27μg 25OHD3/100g (lean meat). Latitude had no effect on the vitamin D3 (P=0.21) or 25OHD3 (P=0.29) content of lean beef, but fat from cattle in the 17° S latitude group contained significantly higher (P<0.01) concentrations of vitamin D3 than fat from the 41° S group of cattle.


Immunocytochemical demonstration of the 'neuronal membrane marker' the D2-glycoprotein in human embryos.
The regional distribution and cellular localization of the D2-glycoprotein in human embryonic tissues have been investigated by immunocytochemistry at the light microscope level in sections of human embryonic brain, spinal cord, some neural crest derivatives, muscle, liver and skin. the majority of neuronal membranes exhibited a positive staining reaction for D2 already in the earliest (15 mm crown-rump length) embryo examined. By 20 mm crown-rump length and later a positive staining associated with membranes of embryonic muscle, bile canaliculi, epithelial cells of the skin and choroid plexus was also observed. The D2 which is immunochemically related to the neuronal cell adhesion molecule (N-CAM), is supposed to be involved in cell adhesion phenomena. We find this suggested function less compatible with demonstrated localization of the D2-glycoprotein.


Genome-wide identification and expression profiling analysis of trihelix gene family in tomato.
The trihelix family, classified as GT factors due to their binding specificity for GT elements, constitutes a plant-specific transcription factor family with a conserved trihelix DNA binding domain. In the present study, the comprehensive analysis of 36 putative GT factors was performed in tomato. SlGT members can be classified into six subgroups (GT-1, GT-2, SH4, SIP1, GT-γ and GT-δ). Expression analysis of SlGT gene transcripts showed the distinct expression patterns of SlGT genes in various tomato organs. All the SlGT genes were regulated in response to various abiotic stresses and hormone treatments by the quantitative real-time PCR (qRT-PCR) analysis. Several SlGT genes, including SlGT-27 and SlGT-34, were highly regulated by multiple abiotic stresses and phytohormone treatments. Taken together, our results presented here would be providing a useful platform for molecular clone and functional identification of SlGT genes in tomato.


Detection and attribution of the start of the growing season changes in the Northern Hemisphere.
Global climate change has led to significant changes in land surface phenology. At present, research on the factors influencing the start of the growing season (SOS) mainly focuses on single factor effects, such as temperature and precipitation, ignoring the combined action of multiple factors. The impact of multiple factors on the spatial and temporal patterns of the SOS in the Northern Hemisphere is not clear, and it is necessary to combine multiple factors to quantify the degrees of influence of different factors on the SOS. Based on the GIMMS3g NDVI dataset, CRU climate data and other factor data, we used geographic detector model, random forest regression model, multiple linear regression, partial correlation analysis and Sen + Mann-Kendall trend analysis to explore the variation of the SOS in the Northern Hemisphere to reveal the main driving factors and impact threshold of 17 influencing factors on the SOS. The results showed that (1) during the past 34 years (1982-2015), the SOS in Europe and Asia mainly showed an advancing trend, whereas the SOS in North America mainly showed a delaying trend. (2) The SOS was mainly controlled by frost frequency, temperature and humidity. Increasing frost frequency inhibited the advancement of the SOS, and increasing temperature and humidity promoted the advancement of the SOS. (3) There were thresholds for the influences of the driving factors on the SOS. Outside the threshold ranges, the response mechanism of the SOS to driving factors changed. The results are important for understanding the response of the SOS to global climate change.


Lyme disease spirochetes and ixodid tick spirochetes share a common surface antigenic determinant defined by a monoclonal antibody.
Ixodid tick-associated spirochetes have been implicated as the etiological agents of Lyme disease. We raised a murine monoclonal antibody (H5332) against a spirochete, strain B31, isolated from Ixodes dammini ticks. In indirect immunofluorescence assays and western blot analyses, H5332 reacted with whole cells or isolated components of not only strain B31 but also spirochetes isolated from Ixodes ricinus ticks, a field mouse, a raccoon, and patients with Lyme disease. In contrast, H5332 did not bind to representative borreliae, treponemes, and leptospires. Using indirect immunofluorescence assays and immune electron microscopy, we found the H5332 determinant to be diffusely distributed over the surface of prefixed spirochetes but to be aggregated in patches when the organisms were incubated with H5332 and a second ligand before fixation. Radioimmunoprecipitation and western blot studies revealed the H5332 determinant to be either on or tightly associated with an abundant outer membrane protein with an apparent subunit molecular weight of 31,000.


Anaerobic digestion of cattail by rumen cultures.
The anaerobic digestion of aquatic plants could serve the dual roles for producing renewable energy and reducing waste. In this study, the anaerobic digestion of cattail (Typha latifolia linn), a lignocellulosic aquatic plant, by rumen microorganisms in batch cultures was investigated. At a substrate level of 12.4 g/l volatile solids (VS) and pH 6.7, maximum VS conversion of 66% was achieved within an incubation time of 125 h. However, a decrease in pH from 6.7 to 5.8 resulted in a marked reduction in VS conversion. The total volatile fatty acids (VFAs) yield was about 0.56 g/g VS digested. Acetate and propionate were the major aqueous fermentation products, while butyrate, i-butyrate and valerate were also formed in smaller quantities. Biogas that was produced was composed of carbon dioxide, methane and hydrogen. A modified Gompertz equation was developed to describe substrate consumption and product formation. The hydrolysis of insoluble components was the rate-limiting step in the anaerobic digestion of cattail.


Effectiveness-implementation hybrid type 2 trial evaluating two psychoeducational programmes for severe hypoglycaemia in type 1 diabetes: implementation study protocol.
Two of the most acute and feared complications in type 1 diabetes (T1D) are hypoglycaemia and severe hypoglycaemia (SH). While impaired awareness of hypoglycaemia (IAH) can lead to SH with cognitive and motivational barriers implicated, the available education does not integrate behavioural change techniques to address these. A novel Hypoglycaemia Awareness Restoration Programme despite optimised care (HARPdoc) is currently being tested against an established blood glucose awareness training (BGAT) within a parallel, two-arm, group randomised, blinded trial (with its own protocol; NCT02940873) with adults with T1D whose problems with hypoglycaemia and SH have persisted despite otherwise optimised insulin management. While both programmes are aimed at reducing hypoglycaemia, SH and IAH, it is the former that integrates behavioural change techniques.The aim of the current (implementation) study is to evaluate delivery of both HARPdoc and BGAT and explore associations between implementation outcomes and trial endpoints; as well as to develop an evidence-based implementation blueprint to guide implementation, sustainment and scale-up of the effective programmes. Guided by the implementation science tools, frameworks, methods and principles, the current study was designed through a series of focus groups (n=11) with the key intervention stakeholders (n=28)-including (1) individuals with lived experience of T1D, IAH and a pilot version of the HARPdoc (n=6) and (2) diabetes healthcare professionals (n=22). A mixed-methods approach will be used throughout. Stakeholder engagement has underpinned study design and materials to maximise relevance, feasibility and impact. The protocol has been reviewed and received ethical approval by the Harrow Research Ethics Committee (18/LO/1020; 240752) on 1 October 2018. The findings will be submitted to a peer-reviewed journal and presented at scientific meetings. NCT02940873; Pre-results.


Biosynthesis and cytoplasmic accumulation of a chlorinated catechol pigment during 3-chlorobenzoate aerobic co-metabolism in Pseudomonas fluorescens.
A strain of Pseudomonas fluorescens was capable of co-metabolizing 3-chlorobenzoic acid with the production of a chlorinated catechol black pigment. A peroxidase and another enzymatic activity referred to as a polyphenol oxidase were found to be involved in the oxidation of 4-chlorocatechol to 4-chloro-1,2-benzoquinone, i.e. in the production of highly reactive substrates for pigment formation. Therefore, P. fluorescens cells were seen to take an active part not only in 3-chlorobenzoate mineralization but also in overall pigment production. pH was found to be a key parameter in the regulation of the activity of P. fluorescens oxidoreductive enzymes. Ultrastructural investigations showed that electron dense granules of pigment were distributed throughout the cytoplasm of Pseudomonas fluorescens cells grown in presence of 3-chlorobenzoate, as confirmed also by Thiéry cytochemical investigations. In these cells, an extensive contraction of the cytoplasm as well as a significant damage to the cell wall after two days of incubation, suggested that pigment production caused a premature death of the cells accompanied by the leakage of the cell content. Pigment production seemed to occur mostly in the cytoplasmic context where the electron dense material accumulates until it is released in the medium after the cell lysis.


Automatic REM detection: modifications on an existing system and preliminary normative data.
This paper describes hardware changes and additions to a previously reported sleep rapid eye movement (REM) automatic detection system. Specifically, it describes the design philosophy of a new and optimum analogue bandpass prefilter, new detection criteria based on a detailed study of the waveform distortion due to AC coupling and bandpass prefiltering and the implementation of an artifact detection system for a more accurate detection of seemingly REM-related electro-oculographic (EOG) waveforms. Preliminary normative data on phasic REM patterns from young adults, detected by the described system, are also presented.


Retention of small bowel capsule endoscopy.
Capsule endoscopy is widely recognized as a safe and effective procedure to evaluate the small bowel and/or the colon noninvasively. Although infrequent, capsule retention is the most feared adverse event related to this technique. A better knowledge of risk factors, improvement of patients' selection and precapsule patency assessment may further contribute to reducing the incidence of capsule retention, even in patients at increased risk for this complication. This review addresses the main risk factors for capsule retention, strategies for risk reduction such as patients' selection, dedicated cross-sectional imaging and rational use of the patency capsule, as well as management options and outcomes in the case of capsule retention. Capsule retention is infrequent, and it is usually manageable conservatively with favorable clinical outcomes. Patency capsules and dedicated small-bowel cross-sectional techniques such as CT or MR enterography should be used selectively and are both effective in decreasing the rate of capsule retention. However, none of them can completely eliminate the risk of retention.


Sequential biases in accumulating evidence.
Whilst it is common in clinical trials to use the results of tests at one phase to decide whether to continue to the next phase and to subsequently design the next phase, we show that this can lead to biased results in evidence synthesis. Two new kinds of bias associated with accumulating evidence, termed 'sequential decision bias' and 'sequential design bias', are identified. Both kinds of bias are the result of making decisions on the usefulness of a new study, or its design, based on the previous studies. Sequential decision bias is determined by the correlation between the value of the current estimated effect and the probability of conducting an additional study. Sequential design bias arises from using the estimated value instead of the clinically relevant value of an effect in sample size calculations. We considered both the fixed-effect and the random-effects models of meta-analysis and demonstrated analytically and by simulations that in both settings the problems due to sequential biases are apparent. According to our simulations, the sequential biases increase with increased heterogeneity. Minimisation of sequential biases arises as a new and important research area necessary for successful evidence-based approaches to the development of science. © 2015 The Authors. Research Synthesis Methods Published by John Wiley & Sons Ltd.


The risk of progression of lentigo maligna to lentigo maligna melanoma.
An analysis is presented which estimates the risk of progression of lentigo maligna (LM) to lentigo maligna melanoma (LMM) in U.S. whites using three data sources: the Health and Nutrition Examination Survey I for estimation of the age-specific prevalence of LM; the Surveillance, Epidemiology, and End Results Program for estimation of the age-specific incidence of melanoma; and the data from three melanoma registries for estimation of the age-specific case fraction of LMM among all invasive melanomas. The risk varies with age and is likely to be greater than estimated here for patients who present themselves for evaluation of changes in a lesion of LM. Our analysis suggests that the risk of progression from LM to LMM is substantially lower than is commonly believed.


Risk factors for pneumococcal carriage in adults living with HIV on antiretroviral therapy in the infant pneumococcal vaccine era in Malawi.
Adults living with HIV (ALWHIV) on antiretroviral therapy (ART) are at high risk of pneumococcal carriage and disease. To help evaluate carriage risk in African ALWHIV at least 4 years after infant pneumococcal conjugate vaccination introduction in 2011, we assessed association between pneumococcal carriage and potential risk factors. Nasopharyngeal swabs were collected from adults aged 18-40 years attending an ART clinic during rolling, cross-sectional surveys in Blantyre, Malawi between 2015 and 2019. We fitted generalized additive models to estimate the risk of sex, social economic status (SES), living with a child less than 5 years, and ART duration on carriage. Of 2067 adults, median age was 33 years (range 28-37), 1427 (69.0%) were women, 1087 (61.4%) were in low-middle socioeconomic-status (SES), 910 (44.0%) were living with a child less than 5 years, and median ART duration was 3 years (range 0.004-17). We estimated 38.2 and 60.6% reductions in overall and vaccine-serotype carriage prevalence. Overall carriage was associated with low SES, living with a child less than 5 years and shorter duration on ART. By contrast, vaccine-type carriage was associated with living without a child less than 5 years and male sex. Despite temporal reductions in overall and vaccine-serotype carriage, there is evidence of incomplete vaccine-serotype indirect protection. A targeted-vaccination campaign should be considered for ALWHIV, along with other public health measures to further reduce vaccine-serotype carriage and therefore disease.


Ecotoxicological effects of earthworm following long-term Dechlorane Plus exposure.
Dechlorane Plus (DP), similar to persistent organic pollutants, has been widely detected in environmental matrices, especially in sediment and soil. In this study, earthworms Eisenia fetida were exposed to 0.1, 0.5, 6.25 and 12.5 mg kg(-1) DP for 28 d. Lethality, oxidative stress, neurotoxicity and cellulase of E. fetida were assessed to investigate ecotoxicological effects of DP after long-term exposure. Results showed that the direct toxicity of DP was very low. However, death rate, as well as SOD activity, together with changes in activities of CAT, GSH-Px, and GSH levels, indicating that oxidative stress may play a significant role in DP exposure. In addition, DP also changes the AChE and cellulase activity of earthworms even under low DP concentration after long-term exposure. Moreover, comet assay results showed that DP exposure increased the levels of tDNA significantly (p < 0.05) even in the lowest treatment (0.1 mg kg(-1) DP). Combined with the results of enzyme activity, oxidative damage and comet assay, it can be suggested that earthworms experience more stress of DP during long-time exposure. This study provides insight into the toxicological effects of DP on earthworm model, and may be useful for risk assessment of DP on soil ecosystems.


The management of Paget's disease of the breast.
Paget's disease of the breast is a relatively rare condition, accounting for 1% to 3% of primary breast cancers. It is associated with an underlying carcinoma, invasive or noninvasive (ductal carcinoma in situ), in most of the cases. Primary treatment is surgical with adjuvant therapy being dictated by the stage and nature of the underlying tumor. Modified radical mastectomy is the standard of care with breast conservation appropriate in a select group of patients with Paget's disease. This select group includes patients that are diagnosed with nipple-areola changes alone without evidence of a palpable mass or mammographic abnormality. In this group of patients, breast conservation offers local recurrence rates comparable to rates in patients with invasive or noninvasive cancers. In patients diagnosed with associated palpable masses or mammographic abnormalities suggestive of cancer, the recurrence rates are higher and mastectomy is warranted.


Functional and bioactive properties of Larimichthys polyactis protein hydrolysates as influenced by plasma functionalized water-ultrasound hybrid treatments and enzyme types.
The effects of plasma functionalized water (PFW) and its combination with ultrasound (UPFW) on the functional and bioactive properties of small yellow croaker protein hydrolysates (SYPHs) produced from three enzymes were investigated. Fluorescence and UV-Vis spectroscopy indicated that SYPHs tended to unfold with increasing intensity and shift in wavelengths to more flexible conformations under PFW and UPFW treatments. Particle size distribution and microstructure analysis revealed that treatments could disrupt aggregation of protein molecules to increase the roughness, specific surface area, and decrease the particle size of peptides during hydrolysis. The partially denatured structure of SYPHs induced by treatments increased the susceptibility of the fish proteins to exogenous enzymes, thereby accelerating the hydrolytic process to yield peptides with improved solubility, decreased emulsifying and foaming properties, and improved enzyme-specific antioxidant properties. The results revealed that the functionality of SYPHs was influenced by the treatment method and the enzyme type employed.


Optimal office blood pressure levels in younger (<50 years old) Korean hypertensive patients: a nationwide cohort study in South Korea.
It is unclear what thresholds and targets of office blood pressure (BP) for treatment are appropriate in younger (<50 years old) hypertensive patients. Thus, the aim of this study was to evaluate associations of office BP levels with major cardiovascular events (MACEs) in these patients. Using the Korean National Health Insurance Service database, data of 98 192 younger (<50 years old) hypertensive patients having BP measurements available without any history of cardiovascular events from 2002 to 2011 were extracted. This cohort study evaluated associations of BP levels (<120/<70, 120-129/70-79, 130-139/80-89, 140-149/90-99, and ≥150/≥100 mmHg) with MACEs. The study outcome was MACE, a composite of cardiovascular death, myocardial infarction, stroke, and heart failure. In all patients, those treated with antihypertensive medication accounted for 34.7% and those who achieved BP less than 130/80 mmHg accounted for 35.5%. During a mean follow-up of 9.5 ± 2.8 years, 4918 (5%) MACEs were documented in our cohort. The risk of MACE was the lowest [adjusted hazard ratio: 0.77, 95% confidence interval (CI) 0.66-0.89] for those with BP level of less than 120/less than 70 mmHg. It was the highest (hazard ratio 2.0, 95% CI 1.83-2.19) for those with BP level of at least 150/at least 100 mmHg in comparison with those with BP level of 130-139/80-89 mmHg. These results were consistent for all age groups (20-29, 30-39, and 40-49 years) and both sexes. Elevated BP level from less than 120 mmHg/less than 70 mmHg is significantly correlated with an increased risk of MACE in younger (<50 years old) Korean hypertensive patients. Lowering BP to less than 120 mmHg/less than 70 mmHg is needed for these patients.


2-D contour perception resulting from kinetic occlusion.
Kinetic occlusion, the progressive deletion or accretion of texture elements as one surface covers or uncovers another, has been shown to be an important source of information for determining depth order. In the present study, the importance of this information for 2-D contour perception was examined. In Experiment 1, subjects were asked to discriminate four different target shapes defined solely by kinetic occlusion. Discrimination increased with an increase in texture density and velocity, with density as the major factor. In Experiment 2, the targets were defined by static untextured regions as well as by kinetic occlusion. Overall, accuracy was similar to that found in Experiment 1, indicating that the presence of static information had little impact on accuracy. In Experiment 3, subjects were unable to discriminate among the four targets when presented with static versions of the displays used in Experiment 2. The results from these experiments indicate that kinetic occlusion can be used for discrimination of different 2-D shapes and that density has a more important role in determining accuracy than velocity.


Prenatal diagnosis and genetic counseling of cystic fibrosis.
The number of DNA markers at the chromosome 7-locus of the putative CF gene has markedly increased in recent years, also as a result of intensive research into the possible "candidate" gene. In studies of families with one or more affected children, 97.5% of families are now fully informative, allowing a prenatal diagnosis by chorionic villus sampling in the 10th week and DNA-analysis, which will usually give now a diagnosis with a remaining risk of less than 1%. The microvillar enzyme test in amniotic fluid after amniocentesis in the 18th week will remain an alternative for couples who have a high prior risk, but are either not informative at DNA analysis, or where no information on a (deceased) index case (previous affected child) is available. The risk for a wrong classification is in the order of a few percent (in a 1:4 prior risk case) and careful discussion of the limitations are needed when this test is applied to cases with a lower prior risk. The linkage disequilibrium established for a number of RFLP's (Restriction Fragment Length Polymorphisms), as detected by various probes and various restriction enzymes around the CF locus has opened the possibility to refine the risk estimation of heterozygosity for individuals outside families with CF-affected children. The presence of certain haplotypes may change the risk for being CF heterozygote from +/- 1:7 to 1:250, as compared to a population risk of +/- 1:25.(ABSTRACT TRUNCATED AT 250 WORDS)


Colon preneoplasia after carcinogen exposure is enhanced and colonic serotonergic system is suppressed by food deprivation.
Calorie restriction regimens usually promote health and extend life-span in mammals. This is partially related to their preventive effects against malignancies. However, certain types of nutritional restriction failed to induce beneficial effects. The American Institute of Nutrition defines calorie restriction as diets which have only 40% fewer calories, but provide normal amounts of necessary food components such as protein, vitamins and minerals; whereas, food restriction means 40% less of all dietary ingredients plus 40% less calories. Our study aimed to test the hypothesis that the latter type of food deprivation (40% less food than consumed by standard fed rats) might increase cancer risk instead of reducing it, as is generally assumed for all dietary restrictive regimens. Since the endogenous modulation of the colon serotonergic system has been observed to play a role during the early steps of carcinogenesis we also investigated whether the serotoninergic system could be involved in the food intake modulation of cancer risk. For this, rats were exposed to a carcinogen and subjected to food deprivation for 56 days. Triglyceride levels and visceral adipose tissue were reduced while hepatic and colonic lipid peroxidation was increased. This dietary restriction also decreased serotonin levels in colon, and gene expression of its intestinal transporter and receptors. Finally, the numbers of preneoplastic lesions in the colon tissue of carcinogen-exposed rats were increased. Our data suggest that food deprivation enhances formation of early tumorigenic lesions by suppressing serotonergic activity in colon tissue.


Chromosomal anomalies in patients with retinoblastoma.
Karyotypes from 50 persons with retinoblastoma confirmed by histopathological examination were studied by conventional staining and Giemsa-banding techniques. Two chromosomal anomalies were found. An interstitial deletion of the long arm of No. 13 chromosome was identified by Giemsa-banding in the karyotypes from a boy with unilateral retinoblastoma. Another boy with unilateral tumor had a karyotype of 47,XXY. These findings provide additional evidence that a deletion of chromosome No. 13, most likely involving band 13q14, is associated with the development of retinoblastoma. In conjunction with other reports, our findings also suggest that retinoblastoma may be found more frequently in children with chromosomal aneuploidy.


Apoptosis and nutrition: Involvement of amino acid transport system in repression of hybridoma cell death.
Mouse hybridoma cells cultured on the verge of starvation-induced apoptosis, i.e. in a medium diluted with saline, proved to serve as a sensitive screening system for apoptosis-suppressing activity of nutrient medium components. Conventional amino acid mixtures were found to suppress the starvation-induced apoptosis, whereas a vitamin mixture was ineffective. (Franěk F (1995) Biotechnol. Bioeng. 45: 86-90). Recent experiments showed that suppression of apoptosis, and concurrent resumption of growth, could be achieved by addition of single substances at millimolar concentrations. The set of active substances included certain coded L-amino acids (glycine, alanine, serine, threonine, proline, asparagine, glutamine, histidine), non-coded amino acids (β-alanine, taurine, 4-aminobutyric acid), and a non-metabolizable analogue (2-aminoisobutyric acid). This finding shows that some amino acids do not act solely as nutrients, but also as specific signal molecules. The specificity of the effect points to the involvement of adaptively regulated amino acid transport systems A and N in maintaining the balance between triggering and suppression of starvation-induced apoptosis.