The significance of homovanillic acid and 3,4-dihydroxyphenylacetic acid concentrations in human lumbar cerebrospinal fluid.
The concentrations of the acidic dopamine (DA) catabolites homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC) measured in human CSF are supposed to reflect the "turnover" of DA in the brain. The notion of "turnover" is, however, not synonymous with impulse nerve activity in the dopaminergic systems. Significant amounts of DOPAC and HVA could, indeed, be demonstrated in brain structures wherein dopaminergic innervation has not been documented. It must also be noted that DA is not only a neurotransmitter itself, but also a precursor of norepinephrine and epinephrine. Furthermore, in lumbar CSF, levels of biogenic amine catabolites partially reflect metabolism in the spinal cord and may have limited relevance to neurotransmission in the brain. To elucidate these points further, we determined the concentrations of DOPAC and HVA in 22 areas of six human brains and eight levels of six human spinal cords. The data were correlated with the concentration of DA. Quantitative determinations were done using HPLC with electrochemical detection, after solvent and ion-pair extraction. In this study, significant amounts of both DOPAC and HVA were demonstrated in brain structures not previously associated with dopaminergic innervation. The relatively lower DA concentration in these structures suggests that in these regions, the DOPAC and HVA concentrations are unrelated to dopaminergic neurotransmission. The possible role of capillary walls and glial cells in the catabolism of DA must be further evaluated. The demonstration of DOPAC and HVA in the spinal cord is another argument against the hypothesis that CSF levels of HVA and DOPAC reflect closely the activity of the dopaminergic systems in the brain.


Attitudes of health care professionals towards interprofessional teamwork in Ashanti Region, Ghana.
Interprofessional collaboration ensures that high-quality health care is provided leading to improved health outcomes and provider satisfaction. Assessing the attitudes of health care professionals towards teamwork in Ghana is novel. To examine the attitudes of health care professionals towards interprofessional teamwork and assess specific attributes influencing these attitudes in the Ashanti region, prior to implementing an in-service interprofessional HIV training programme. A cross-sectional pre-training online survey using a modified Attitudes Toward Health Care Teams Scale was conducted among health care practitioners undergoing a two-day interactive interprofessional HIV training in Kumasi and Agogo from November 2019 to January 2020. Trainees were diverse health professional cadres selected from five hospitals in the Ashanti region of Ghana. Data was summarised using the mean and standard deviation for continuous variables, and frequencies and percentages for categorical variables. An exploratory factor analysis was conducted to categorise the 14 items of the modified attitudes scale. The Wilcoxon rank-sum (Mann-Whitney) and Kruskal-Wallis tests were used to test the mean attitude difference among the demographic characteristics. Statistical significance was set at p < 0.05. Altogether, 302 health professionals completed the survey. The ages ranged from 20-58 years, mean age 27.96 years (standard deviation 5.90 years). Up to 95% of the trainees agreed with the 14 statements on the modified attitudes scale. Three factors were identified; "quality of care", "team efficiency", and "time constraint" with Cronbach's alpha measures of 0.73, 0.50, and 0.45 respectively. The overall mean attitude score was 58.15 ± 6.28 (95% CI, 57.42-58.88). Attitude of health care professionals towards interdisciplinary teams for patient care varied significantly by age (p = 0.014), health profession cadre (p = 0.005), facility (p = 0.037), and professional experience (p = 0.034). Strengthening in-service interprofessional training for health practitioners especially early career professionals in the Ashanti region would be valuable.


Single-molecule approaches embrace molecular cohorts.
Enormous mechanistic insight has been gained by studying the behavior of single molecules. The same approaches used to study proteins in isolation are now being leveraged to examine the changes in functional behavior that emerge when single molecules have company.


Surgical ablation of atrial fibrillation with a novel bipolar radiofrequency device.
When used for epicardial ablation, unipolar devices do not predictably yield transmural scars. Bipolar radiofrequency proved highly effective on the animal model, but clinical experience is still initial. We describe acute electrophysiologic findings and follow-up results of epicardial ablation with a novel bipolar radiofrequency device. A bipolar ablator was used to perform a simplified left atrial lesion set in 90 consecutive patients with atrial fibrillation undergoing open heart surgery. Pacing thresholds were assessed during surgery to validate 24 pulmonary vein encircling lines (12 patients). Follow-up was 100% complete. In 67 of 90 patients (84%), mitral valve disease was the main indication to surgery. Atrial fibrillation was continuous in 74 patients (82%) and intermittent in 16 patients (18%). Pacing threshold assessment showed a complete conduction block in 22 of 24 pulmonary vein couples (92%) after a single ablation and in all patients after doubling of the encircling lines. No complications related to the ablation procedure were recorded. The sinus rhythm restoration rate was 79% at 3 months, 87% at 6 months, and 89% (17/18 patients) at 1 year. Postablation organized arrhythmias consisted in right atrial flutter in 2 patients (2%) and left atrial flutter in 6 patients (7%). Epicardial ablation with bipolar radiofrequency grants acute transmurality. A simplified lesion set proved highly effective in eliminating atrial fibrillation at 1-year follow-up. Our data suggest that addition of a lesion to the mitral annulus is advisable to prevent left atrial flutter.


Expression Quantitative Trait Methylation Analysis Reveals Methylomic Associations With Gene Expression in Childhood Asthma.
Nasal (airway) epithelial methylation profiles have been associated with asthma, but the effects of such profiles on expression of distant cis-genes are largely unknown. To identify genes whose expression is associated with proximal and distal CpG probes (within 1 Mb), and to assess whether and how such genes are differentially expressed in atopic asthma. Genome-wide expression quantitative trait methylation (eQTM) analysis in nasal epithelium from Puerto Rican subjects (aged 9-20 years) with (n = 219) and without (n = 236) asthma. After the eQTM analysis, a Gene Ontology Enrichment analysis was conducted for the top 500 eQTM genes, and mediation analyses were performed to identify paths from DNA methylation to atopic asthma through gene expression. Asthma was defined as physician-diagnosed asthma and wheeze in the previous year, and atopy was defined as at least one positive IgE to allergens. Atopic asthma was defined as the presence of both atopy and asthma. We identified 16,867 significant methylation-gene expression pairs (false-discovery rate-adjusted P < .01) in nasal epithelium from study participants. Most eQTM methylation probes were distant (average distance, ∼378 kb) from their target genes, and also more likely to be located in enhancer regions of their target genes in lung tissue than control probes. The top 500 eQTM genes were enriched in pathways for immune processes and epithelial integrity and were more likely to have been previously identified as differentially expressed in atopic asthma. In a mediation analysis, we identified 5,934 paths through which methylation markers could affect atopic asthma through gene expression in nasal epithelium. Previous epigenome-wide association studies of asthma have estimated the effects of DNA methylation markers on expression of nearby genes in airway epithelium. Our findings suggest that distant epigenetic regulation of gene expression in airway epithelium plays a role in atopic asthma.


Immunoepidemiology of Wuchereria bancrofti infection in two East African communities: antibodies to the microfilarial sheath and their role in regulating host microfilaraemia.
The response pattern of specific antibodies to the microfilarial sheath (sheath-Ab) of the mosquito-borne filarial parasite Wuchereria bancrofti was investigated in individuals from two East African communities with different levels of endemicity. Individuals from both communities presented a strong inverse relationship between positivity for sheath-Ab and being positive for microfilariae (mf) and circulating filarial antigens (CFA). The prevalence of sheath-Ab positivity was highest in young individuals, but peaked at a younger age in the high (1-14 years) than the low (15-19 years) endemicity community. IgG1, IgG2, IgG3 and IgE intensities to a crude adult filarial worm antigen were higher, and IgG4 intensities were lower, in sheath-Ab positive than in sheath-Ab negative individuals, probably reflecting the infection status of individuals. From the study it appears that individuals become sheath-Ab positive before mf and/or CFA can be detected in the peripheral blood, and only after later disappearance of sheath-Ab from the circulation can CFA and mf be diagnosed. In light of the findings, possible roles of the distinct sheath-Ab in the host-parasite relationship are discussed, and a hypothesis is proposed which suggests that sheath-Ab play an important role in the regulation of host microfilaraemia.


Effect of neomycin on the hydrolysis and toxicity of vicine and convicine in rats.
This study in the rat established the effects that a broad-spectrum and poorly absorbed antibiotic, neomycin sulfate, had on the in vitro and in vivo hydrolysis of vicine and convicine by the intestinal microflora, and on vicine- and convicine-induced depletion of blood glutathione and the resulting toxicity. The in vitro studies demonstrated that digesta from the caecum and large intestine were highly effective in hydrolysing vicine and convicine, whereas digesta from the same sections of the gastro-intestinal tract of neomycin-treated rats were much less effective (P < 0.0001). The in vivo studies showed that the total amount of vicine and convicine excreted in the urine and faeces was much greater in neomycin-treated rats compared with controls (P < 0.05), indicating the ability of neomycin to increase the amount of glycosides, particularly that of vicine, excreted in the faeces. The ability of glycosides to decrease the concentration of glutathione in blood (P < 0.05) and to increase rat mortality was greatly reduced in rats that were treated with neomycin, particularly in those treated ip with the toxin. Thus, the results demonstrated that neomycin reduced the rate at which vicine and convicine were hydrolysed in the lower section of the gastro-intestinal tract, and that neomycin treatment was associated with a reduced toxicity of the glycosides.


[Results of macula hole surgery in Iceland 1996-2002].
To evaluate retrospectively the results of macular hole surgery in Iceland 1996-2002. 25 patients underwent macular hole surgery in this time period. Data was obtained from hospital records of all those patients. Best corrected vision vas measured before and after surgery. The staging of the macular hole was evaluated before surgery and anatomical closure after surgery was recorded. Any adjuvants used during the operation were noted, and determined if there were any differences in outcomes depending on which adjuvants were used. After one operation the anatomical success was 72% and 79% after two surgeries. Visual acuity improved > or = 2 lines on the Snellen card in 11 of 29 eyes (38%); vision stayed the same (did not improve or get worse of more then > or = 2 lines) in 16 of 29 eyes (55%), and deteriorated > or = 2 lines in two of 29 eyes (7%). There were no significant clinical differences in which adjuvant therapy was used. Anatomical success of macular hole surgery in Iceland was comparable with results in foreign studies. The visual outcome was significantly better after surgery, but those results were not as good as in foreign studies.


Comparison between first and second generation tests to determine the frequency of anti-HCV antibodies in uremic patients in replacement dialytic therapy.
Research was carried out on 74 hemodialysis patients to determine the frequency of anti-HCV antibodies; this was done by means of the 1st and 2nd generation screening and control tests. The 1st generation tests showed 9 HCV-seropositive patients, while the 2nd generation tests demonstrated 15 HCV-seropositive patients. It seems evident that the 2nd generation tests are more sensitive than those of the 1st.


Recombinogenic activity of 10 chemical compounds in male germ cells of Drosophila melanogaster.
The Drosophila melanogaster male germ cells are sensitive to the recombinogenic activity of chemical compounds. In our experiments, we have employed four recessive markers located on the 2nd chromosome: dumpy (dp, 13.0), black (b, 48.5), cinnabar (cn, 57.5), and brown (bw, 104.5); b and cn flank the centromere. Three-day-old larvae, heterozygous for these markers, were treated chronically by oral administration with the test compounds. The chemicals already shown to be positive or negative in the assay systems to test chemical agents in D. melanogaster are three carcinogens (4-nitroquinoline N-oxide, hydroxylamine HCl, and acrylamide), four herbicides (maleic hydrazide, alachlor, trifluralin, and amitrol), and three insecticides (endrin, piperonyl butoxide, and allethrin). In our study, some compounds induced recombinogenic effects in Drosophila premeiotic male germ cells, and comparison of our results with those reported in the literature with the Drosophila wing somatic mutation and recombination assay showed that the somatic cells and the germinal cells have a differential response to the defined compounds.


No relationship between the digit ratios (2D:4D) and salivary testosterone change: Study on men under an acute exercise.
The digit ratio (2D:4D) is said to be a potential marker of exposure to prenatal sex steroids. Some studies suggest that the 2D:4D is also linked with the testosterone response to challenging situations due to organizational effect of prenatal hormonal milieu on adult endocrine functioning. However, up to date, there were only four studies (conducted on small samples) that examined the 2D:4D and the testosterone response to a challenging situation (i.e. physical exertion or aggressive context). Here, we examined the relationship between the 2D:4D and the testosterone change under an acute exercise among 97 men. We found that the digit ratios (the right 2D:4D, the left 2D:4D, and the right minus left 2D:4D) were neither predictors of pre-exercise testosterone, nor the change in testosterone level after a cycling task. Our results add a contradictory to previous studies evidence in a discussion on the links of the 2D:4D and the testosterone change.


Permanent cardiac pacing after a cardiac operation: predicting the use of permanent pacemakers.
The need for permanent cardiac pacing after cardiac operations is infrequent but associated with increased morbidity and resource utilization. We identified patient risk factors for pacemaker insertion to enable development of a predictive model. Data were collected prospectively for 10,421 consecutive patients who had cardiac operations between January 1990 and December 1995. Two hundred fifty-five patients (2.4%) were identified as having received a permanent pacemaker during the same hospitalization. Logistic regression analysis was performed to determine the independent, multivariate predictors of permanent pacing. The predictive accuracy and precision of the logistic regression model was evaluated in the 1996 database of 2,236 consecutive patients by the calculation of Brier scores. Eight independent predictors of permanent pacemaker requirement were identified. The factor-adjusted odds ratios (OR) with 95% confidence interval (CI) associated with each predictor are as follows: (1) valve replacement surgery (aortic: OR 5.8, CI 3.9-8.7; mitral: OR 4.9, CI 3.1-7.8; tricuspid: OR 8.0, CI 5.5-11.9; double: OR 8.9, CI 5.5-14.6; and triple: OR 7.5, CI 2.9-19.3); (2) repeat operation: OR 2.4, CI 1.8-3.3; (3) age 75 years or older: OR 3.0, CI 2.0-4.4; (4) ablative arrhythmia operation: OR 4.2, CI 1.9-9.5; (5) mitral valve annular reconstruction: OR 2.4, CI 1.4-4.2; (6) use of cold blood cardioplegia: OR 2.0, CI 1.2-3.6; (7) preoperative renal failure: OR 1.6, CI 1.0-2.6; and (8) active endocarditis: OR 1.7, CI 0.9-3.0. A model for postoperative permanent pacemaker requirement using the eight predictors was formulated and tested (Brier score = 0.017+/-0.003; Z = 0.18). The proposed predictive model correlated highly with actual pacemaker use, which suggests that the requirement for pacing results from either operative trauma or increased ischemic burden. Preoperative identification of patients at increased risk of conduction disturbances may allow for earlier detection and improved treatment. Patients requiring postoperative pacing had increased morbidity and length of stay.


Critical Analysis of the Causes of In-Hospital Mortality following Colorectal Resection: A Queensland Audit of Surgical Mortality (QASM) Registry Study.
Colorectal resection is a major gastrointestinal operation. Improvements in peri-operative care has led to improved outcomes; however, mortalities still occur. Using data from the Queensland Audit of Surgical Mortality (QASM), this study examines the demographic and clinical characteristics of patients who died in hospital following colorectal resection, and also reports the primary cause of death in this population. Patients who died in hospital following colorectal resection in Queensland between January 2010 and December 2020 were identified from the QASM database. There were 755 patients who died in the 10 year study period. Pre-operatively, the risk of death as subjectively determined by operating surgeons was 'considerable' in 397 cases (53.0%) and 'expected' in 90 cases (12.0%). The patients had a mean of 2.7 (±1.5) co-morbidities, and a mean American Society of Anaesthesiologists (ASA) score of 3.6 (±0.8). Operations were categorised as emergency in 579 patients (77.2%), with 637 patients (85.0%) requiring post-operative Intensive Care Unit (ICU) support. The primary cause of death was related to a surgical cause in 395 patients (52.7%) and to a medical cause in 355 patients (47.3%). The primary causes of death were advanced surgical pathology (n=292, 38.9%), complications from surgery (n=103, 13.7%), complications arising from pre-existing medical co-morbidity (n=282, 37.6%) or new medical complications unrelated to pre-existing conditions (n=73, 9.7%). Patients who died had significant co-morbidities and often presented emergently with an advanced surgical pathology. Surgical and medical causes of death both contributed equally to the mortality burden.


Stable engineered vascular networks from human induced pluripotent stem cell-derived endothelial cells cultured in synthetic hydrogels.
Here, we describe an in vitro strategy to model vascular morphogenesis where human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) are encapsulated in peptide-functionalized poly(ethylene glycol) (PEG) hydrogels, either on standard well plates or within a passive pumping polydimethylsiloxane (PDMS) tri-channel microfluidic device. PEG hydrogels permissive towards cellular remodeling were fabricated using thiol-ene photopolymerization to incorporate matrix metalloproteinase (MMP)-degradable crosslinks and CRGDS cell adhesion peptide. Time lapse microscopy, immunofluorescence imaging, and RNA sequencing (RNA-Seq) demonstrated that iPSC-ECs formed vascular networks through mechanisms that were consistent with in vivo vasculogenesis and angiogenesis when cultured in PEG hydrogels. Migrating iPSC-ECs condensed into clusters, elongated into tubules, and formed polygonal networks through sprouting. Genes upregulated for iPSC-ECs cultured in PEG hydrogels relative to control cells on tissue culture polystyrene (TCP) surfaces included adhesion, matrix remodeling, and Notch signaling pathway genes relevant to in vivo vascular development. Vascular networks with lumens were stable for at least 14days when iPSC-ECs were encapsulated in PEG hydrogels that were polymerized within the central channel of the microfluidic device. Therefore, iPSC-ECs cultured in peptide-functionalized PEG hydrogels offer a defined platform for investigating vascular morphogenesis in vitro using both standard and microfluidic formats. Human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) cultured in synthetic hydrogels self-assemble into capillary networks through mechanisms consistent with in vivo vascular morphogenesis.


Chemical inducers of dimerization: the atomic structure of FKBP12-FK1012A-FKBP12.
A ligand that simultaneously binds two proteins must have two high affinity protein binding domains joined in a fashion that facilitates, or at least does not prevent, protein-protein interaction. Designing such ligands is challenging, and a high resolution X-ray structure of FKBP12-FK1012A-FKBP12 illustrates the subtleties of one successful design.


Cerebral processing of nonverbal affective stimuli: differential effects of cognitive and affective sets on hemispheric asymmetry.
The cerebral processing of emotion has been the subject of a number of recent studies, but results relating to hemispheric asymmetry have been quite mixed. The present study attempted to clarify and extend findings in this area by examining the differential effects of cognitive and affective sets on hemisphere asymmetry under conditions of nonverbal emotional stimulation. Electroencephalographic (EEG) data, recorded bilaterally, showed that differential hemispheric processing interacted with emotion (positive or negative), condition (cognitive or affective), and sex. In those emotion/condition combinations where hemispheric asymmetry was observed, there was greater activity in the left hemisphere than in the right. In addition, positive affect produced greater asymmetry than negative, and females showed greater activity in response to positive emotional stimuli than did males. Further, affective conditions yielded higher levels of activity than did cognitive. Finally, electrodermal activity showed lateralization effects as a differential function of cognitive and affective conditions.


The Effects of the Civility, Respect, and Engagement in the Workplace (CREW) Program on Social Climate and Work Engagement in a Psychiatric Ward in Japan: A Pilot Study.
Good social climate and high work engagement are important factors affecting outcomes in healthcare settings. This study observed the effects of a program called Civility, Respect, and Engagement in the Workplace (CREW) on social climate and staff work engagement in a psychiatric ward of a Japanese hospital. The program comprised 18 sessions installed over six months, with each session lasting 30-min. Participation in the program was recommended to all staff members at the ward, including nurses, medical doctors, and others, but it was not mandatory. A serial cross-sectional study collected data at four time-points. Nurses (n = 17 to 22), medical doctors (n = 9 to 13), and others (n = 6 to 10) participated in each survey. The analysis of variance was used to evaluate the changes in the following dependent variables, the Essen climate evaluation schema (EssenCES), the CREW civility scale, and the Utrecht work engagement scale (UWES) over time. We found no significant effects. The effect size (Cohen's d) for EssenCES was 0.35 from baseline to post-installation for all staff members. Effect sizes for EssenCES for medical doctors and UWES for nurses were 0.79 and 0.56, respectively, from baseline to post-program. Differences in social climate and work engagement among Japanese healthcare workers between the baseline and post-installation of the CREW program were non-significant.


Different mechanisms of release of vasoactive amines by mast cells occur in T cell-dependent compared to IgE-dependent cutaneous hypersensitivity responses.
It has recently been shown that the classical 24-48-h component of delayed-type hypersensitivity (DTH) in mice is preceded by an early 2-h component. This early component of DTH resembles IgE-dependent hypersensitivity reactions, but is mediated by an antigen-specific T cell factor that activates cutaneous mast cells (MC). In the current study of cutaneous hypersensitivity reactions, evidence is presented that serotonin and histamine are released from the granules of MC in IgE-dependent responses, but that T cell activation of MC induces preferential release of serotonin. In T cell-dependent reactions, it was found that serotonin was released under experimental conditions in which MC granules were depleted of serotonin, but catabolism of serotonin in the cytosol was prevented. This suggests that T cells induce differential release of serotonin from non-granule storage sites in MC, such as transport vesicles in the cytoplasm. Morphological observations of MC in IgE-dependent compared to T cell-dependent cutaneous reactions were consistent with this hypothesis. Exocytosis of granules characterized IgE activation and was not found in MC that were activated by T cells, which instead showed mild degranulation accompanied by numerous cytoplasmic vesicles.


Effect of chemical modifications on freeze denaturation of lactate dehydrogenase.
Lactate dehydrogenase (LDH) was chemically ethyl-acetimidated (EA-), dimethyladipimidated (DMA-), carbamylated, acetylated, acetoacetylated, or succinylated in order to alter the ionic charges on the epsilon-amino group of lysine residues. Acetylation, acetoacetylation, and succinylation, which change the positive charge at the lysine side chains to a negative one, inactivated the enzymic activity, but the rest of the modifications exerted no such inactivating effects. The active modified enzymes were subjected to freeze denaturation study, using the enzymic activity as an indication of the degree of the denaturation. The active enzymes were diluted with deionized water and stored in a freezer (-23 degrees C) for 1-3 days. Enzymic activity was assayed immediately after thawing. All the modified enzymes retained their activity even after the 3-day frozen storage, while the control or native enzyme lost its activity within 1 day of storage. Furthermore, the modified LDHs freeze-stored in 0.2 M monosodium glutamate (MSG) or 0.2 M lysine-hydrochloride (Lys-HCl) retained their activity. The cryoprotective effects exerted by the modifications and by 0.2 M MSG seemed to be synergistic, whereas those exerted by the modifications and by 0.2 M Lys-HCl did not. The mechanisms of cryoprotection and freeze denaturation are discussed in relationship with the cryoprotective effect exerted by already known cryoprotectants, such as sucrose or dimethyl sulfoxide.


[Ambivalence, old age and agency: Meaning of age-specific ambivalence for the construction of narrative identity].
Different models of the formation of age identity explain the empirically determined difference between the chronological age and the age which is subjectively perceived by elderly people themselves. From a biographical point of view and on the basis of two maximally contrasting empirical cases, this article investigates the ways ambivalence appears in the narrations of elderly people about personal aging and the narrative strategies of coping with this experience of ambivalence. The results of the analysis indicate that the conceptualization of age identity in the different models should be seen more as descriptions of potential but not necessary forms of coping with age-specific ambivalences.


Visual Interpretation with Three-Dimensional Annotations (VITA): three-dimensional image interpretation tool for radiological reporting.
This paper introduces a software framework called Visual Interpretation with Three-Dimensional Annotations (VITA) that is able to automatically generate three-dimensional (3D) visual summaries based on radiological annotations made during routine exam reporting. VITA summaries are in the form of rotating 3D volumes where radiological annotations are highlighted to place important clinical observations into a 3D context. The rendered volume is produced as a Digital Imaging and Communications in Medicine (DICOM) object and is automatically added to the study for archival in Picture Archiving and Communication System (PACS). In addition, a video summary (e.g., MPEG4) can be generated for sharing with patients and for situations where DICOM viewers are not readily available to referring physicians. The current version of VITA is compatible with ClearCanvas; however, VITA can work with any PACS workstation that has a structured annotation implementation (e.g., Extendible Markup Language, Health Level 7, Annotation and Image Markup) and is able to seamlessly integrate into the existing reporting workflow. In a survey with referring physicians, the vast majority strongly agreed that 3D visual summaries improve the communication of the radiologists' reports and aid communication with patients.


Deep brain stimulation mechanisms: the control of network activity via neurochemistry modulation.
Deep brain stimulation (DBS) has revolutionized the clinical care of late-stage Parkinson's disease and shows promise for improving the treatment of intractable neuropsychiatric disorders. However, after over 25 years of clinical experience, numerous questions still remain on the neurophysiological basis for the therapeutic mechanisms of action. At their fundamental core, the general purpose of electrical stimulation therapies in the nervous system are to use the applied electric field to manipulate the opening and closing of voltage-gated sodium channels on neurons, generate stimulation induced action potentials, and subsequently, control the release of neurotransmitters in targeted pathways. Historically, DBS mechanisms research has focused on characterizing the effects of stimulation on neurons and the resulting impact on neuronal network activity. However, when electrodes are placed within the central nervous system, glia are also being directly (and indirectly) influenced by the stimulation. Mounting evidence shows that non-neuronal tissue can play an important role in modulating the neurochemistry changes induced by DBS. The goal of this review is to evaluate how DBS effects on both neuronal and non-neuronal tissue can potentially work together to suppress oscillatory activity (and/or information transfer) between brain regions. These resulting effects of ~ 100 Hz electrical stimulation help explain how DBS can disrupt pathological network activity in the brain and generate therapeutic effects in patients. Deep brain stimulation is an effective clinical technology, but detailed therapeutic mechanisms remain undefined. This review provides an overview of the leading hypotheses, which focus on stimulation-induced disruption of network oscillations and integrates possible roles for non-neuronal tissue in explaining the clinical response to therapeutic stimulation. This article is part of a special issue on Parkinson disease.


Patient safety in the rehabilitation of children with traumatic brain injury and cerebral palsy.
With the advent of newer and better therapies available, patient safety is emerging as a new topic. Pediatric patient safety is relatively new, in that there are few guidelines available. Safety in children with traumatic brain injury (TBI) given the incidence of TBI is very vital. This is an attempt to identify the key points in TBI.


Action of angiotensin converting enzyme inhibitors in rat brain: interaction with isoproterenol assessed by Fos immunocytochemistry.
Low and high doses of angiotensin converting enzyme (ACE) inhibitors have been shown either to enhance or suppress, respectively, the water intake of rats induced by acute administration of isoproterenol. In order to assess the role and sites of action of angiotensin II (Ang II) in this dual action of ACE inhibitors, rats were administered either low or high doses of enalapril or captopril, followed by isoproterenol, and were sacrificed 1 h later for determination of Fos-like immunoreactivity (FLI) in brain. Isoproterenol induced strong FLI in the magnocellular paraventricular (PVN) and supraoptic (SON) nuclei, and moderate staining along the structures of the rostral wall of the lamina terminalis (LT). Low doses of ACE inhibitors either had no effect or slightly increased FLI along the LT following isoproterenol. Enalapril reduced FLI in some other regions, including the parvocellular PVN. In contrast, high doses of ACE inhibitors abolished FLI along the LT, and reduced FLI in the PVN and SON. Captopril, but not enalapril, induced some FLI in the LT, SON and PVN. The data are discussed in terms of access of ACE inhibitors to the brain, and interactions with structures involved in Ang-related water intake.


Modeling and optimization of the flocculation processes for removal of cationic and anionic dyes from water by an amphoteric grafting chitosan-based flocculant using response surface methodology.
In this study, an amphoteric grafting chitosan-based flocculant (carboxymethyl chitosan-graft-poly(2-methacryloyloxyethyl) trimethyl ammonium chloride, denoted as CMC-g-PDMC) was applied to removal of the anionic and cationic dyes, acid Green 25 (AG25) and Basic Bright Yellow (7GL), from water. Flocculation conditions have been optimized by response surface methodology (RSM) on the basis of central composite design (CCD) using flocculant dosage, initial solution pH and temperature as input variables. The second-order and cubic regression models, which have been both tested by the analysis of variance (ANOVA), were constructed to link the output response (the dye removal factor) with the aforementioned input variables, respectively. The second-order regression model well described the process of AG25 removal, whereas the cubic one is more suitable for that of 7GL. The effects of those variables on the flocculation performance of CMC-g-PDMC for removal of the two dyes containing opposite charges from aqueous solutions have been studied, and the flocculation mechanisms including the interactive effects between various influencing factors have been discussed in detail also.


Regulation of protein kinase C by sphingosine and lysosphingolipids.
Protein kinase C (PKC), a calcium and phospholipid dependent protein kinase C, has emerged as a key element in signal transduction and cell regulation. It is activated by sn-1,2-diacylglycerol (DAG) second messengers and it serves as the receptor for phorbol esters, potent tumor promoters. PKC is now known to occur as a family of isoenzymes sharing similar structural features that allow regulation of activity by calcium, phospholipid, and DAG. In vitro mechanisms of PKC regulation by phospholipid, DAG, and phorbol esters have been studied using mixed micelles of Triton X-100/lipids. PKC activation occurs at physiologic mole fractions of phospholipid and DAG, does not require a bilayer, and appears to occur by a two-step mechanism whereby PKC initially interacts with a phospholipid surface and is then activated by the addition of DAG. Similar methodology has been used to explore the inhibition of PKC by different inhibitors that interact with its regulatory domain. Sphingosine and lysosphingolipids are potent inhibitors of PKC that prevent its interaction with DAG/phorbol esters. These naturally occurring metabolites have been shown to affect PKC activity in different cell systems. Disturbances in sphingolipid metabolism may lead to accumulation of lysosphingolipids with consequent inhibition of PKC. Additionally, these naturally occurring metabolites may have physiologic functions in regulating PKC activity by counteracting the action of DAG. The mechanism of action of sphingosine/lysosphingolipids and their possible physiologic function will be discussed.


Male breast carcinoma. An ultrastructural study.
A case of infiltrating ductal carcinoma of the male breast was studied by electron microscopy. The ultrastructure of the tumor is similar to that described in the female breast. Myofibroblasts are the preponderant cells in the stroma. Their significance and possible relation to hormonal stimuli are discussed.


Cerebral lesions in mice infected with Plasmodium berghei are the result of an immunopathological reaction.
Cerebral lesions in mice with Plasmodium berghei infections can be prevented by timely treatment with immune serum, by splenectomy, and by administration of dexamethasone. T cell deficient mice do not develop cerebral lesions. The results of these studies are compatible with the hypothesis that a pathological reaction is responsible for development of cerebral lesions in mice infected with P. berghei.


[Design, synthesis and characterization of cyanopyrrolidine-bearing compounds as DPP-4 inhibitors].
To synthesize novel cyanopyrrolidine-bearing compounds as dipeptidyl peptidase 4 (DPP4) inhibitors and characterize their biological activities in vitro. Eleven analogues of carbonitrilpyrrolidine were designed and synthesized by substitution reaction of (S)-2-(2-cyanopyrrolidin-1-yl)acetyl bromide with substituted phenyl piperazine pyridazinones. The structures of the compounds were characterized by (1)H-NMR and MS spectra. Biological evaluation indicated that most of the compounds exhibited moderate inhibitory activities against DPP4. The preliminary bioassay indicates that all the synthesized compounds have moderate DPP-4 inhibition activity, especially the compounds 1j and 1k with inhibition rates reaching 26.14% and 34.15% at the concentration of 1×10(5) nmol/L, respectively.


Elucidating Fibroblast Growth Factor-Induced Kinome Dynamics Using Targeted Mass Spectrometry and Dynamic Modeling.
Fibroblast growth factors (FGFs) are paracrine or endocrine signaling proteins that, activated by their ligands, elicit a wide range of health and disease-related processes, such as cell proliferation and the epithelial-to-mesenchymal transition. The detailed molecular pathway dynamics that coordinate these responses have remained to be determined. To elucidate these, we stimulated MCF-7 breast cancer cells with either FGF2, FGF3, FGF4, FGF10, or FGF19. Following activation of the receptor, we quantified the kinase activity dynamics of 44 kinases using a targeted mass spectrometry assay. Our system-wide kinase activity data, supplemented with (phospho)proteomics data, reveal ligand-dependent distinct pathway dynamics, elucidate the involvement of not earlier reported kinases such as MARK, and revise some of the pathway effects on biological outcomes. In addition, logic-based dynamic modeling of the kinome dynamics further verifies the biological goodness-of-fit of the predicted models and reveals BRAF-driven activation upon FGF2 treatment and ARAF-driven activation upon FGF4 treatment.


Staging of intrauterine growth-restricted fetuses.
The purpose of this study was to evaluate the value of cardiovascular, ultrasonographic, and clinical parameters for developing a staging classification of intrauterine growth-restricted (IUGR) fetuses delivered at 32 weeks or earlier. Intrauterine growth restriction was defined as the presence of an estimated fetal weight below the 10th percentile. Intrauterine growth-restricted fetuses were staged according to the following parameters, with the presence of any 1 parameter in a stage placing the fetus in that stage: stage I, an abnormal umbilical artery or middle cerebral artery pulsatility index; stage II, an abnormal middle cerebral artery peak systolic velocity, umbilical artery absent/reversed diastolic flow, umbilical vein pulsation and an abnormal ductus venosus pulsatility index; and stage III, reversed flow at the ductus venosus or reversed flow at the umbilical vein, an abnormal tricuspid E wave (early ventricular filling)/A wave (late ventricular filling) ratio, and tricuspid regurgitation. Each stage was divided into A (amniotic fluid index [AFI] <5 cm) and B (AFI >5 cm). The presence of maternal abnormalities was also reported. Seventy-four IUGR fetuses delivered at 32 weeks or earlier were included. Gestational age at delivery was greater in stage I fetuses compared with the other stages. Birth weight decreased with advancing stages. Stage III fetuses had the lowest AFI. There was a direct correlation between the severity of staging and both perinatal mortality and mortality occurring between 20 weeks' gestation and before the neonates were discharged from the hospital (P < .05). The staging system proposed here may allow comparison of outcome data for IUGR fetuses and may be valuable in determining more timely delivery for these high-risk fetuses.


Composition and subcellular distribution of glycoproteins and glycosaminoglycans undergoing axonal transport in garfish olfactory nerves.
The study examined the subcellular distribution of [3H]glucosamine-labeled glycoconjugates undergoing axonal transport in 100,000 x g soluble and two membranous subfractions of the garfish olfactory nerve. Analysis was made of intact glycoconjugates and of glycopeptides and glycosaminoglycans derived from these molecules by limit protease digestion. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed labeling of a variety of high-molecular-weight molecules with a lower molecular weight distribution in the soluble fraction than in the membranous fractions. Following protease digestion, nearly two-thirds of transported radioactivity in glycopeptides was recovered in the plasma membrane-enriched subfraction, with the remainder equally divided between soluble and higher density membrane fraction. Comparison of the distribution of glycopeptide radioactivity and chemically assayed hexosamine revealed transport labeling of a large variety of different-sized neutral and acidic glycopeptides in all subfractions. Transport labeling of most glycoprotein carbohydrate chains was in proportion of their hexosamine content. Transported glycosaminoglycan label was most heavily concentrated in the plasma membrane fraction, whereas hexosamine was most concentrated in the higher density membrane fraction. The labeling pattern suggested both transported and nontransported pools of these molecules. The specific glycosaminoglycans chondroitin sulfate and heparan sulfate were recovered in all subfractions, whereas hyaluronic acid was confined to the soluble fraction.


Road to elimination of HCV: Clinical challenges in HCV management.
Since its discovery in 1989, the road to a cure for hepatitis C virus (HCV) has been slow, but most patients can now expect to achieve a sustained virological response (SVR). With direct-acting antiviral (DAA) combination therapies such as glecaprevir/pibrentasvir and velpatasvir/sofosbuvir, 98% of patients successfully eradicate the virus, even if previous treatments failed or if resistance-associated substitutions (RASs) are present. Adverse events are rare or mild, and patients with compensated cirrhosis and other co-morbidities are often eligible for treatment. However, a small number of patients fail to eradicate the virus even after retreatment. The cause of failure is mainly due to emergence of NS5A P32 deletion mutants after initial DAA therapy in genotype 1b patients, although the reason is unknown for some patients. Alternative therapies that do not rely on NS5A inhibitors, such as sofosbuvir plus ribavirin, can be attempted in these patients. While scaled-up treatment efforts present a challenge, another problem is that many carriers are unaware of their infection. Long-term damage to the liver becomes irreversible, and patients who are not diagnosed in time can develop liver cancer or liver failure even after eliminating the virus. The long-term costs of treatment of advanced liver disease in undiagnosed patients relative to the immediate costs of DAA therapy should be considered. As no vaccine is yet available, eventual elimination of the virus requires identifying and treating undiagnosed cases and screening of high-risk populations such as injection drug users and men who have sex with men and female sex workers.


Alz-50/Gallyas-positive lysosome-like intraneuronal granules in Alzheimer's disease and control brains.
The Gallyas-Braak silver impregnation method revealed neurons containing well-defined intraneuronal granules in both Alzheimer's disease and normal control brains. The granules were immunostained prominently with the Alz-50 antibody and, to a lesser degree, with the tau-2 antibody, but not with other anti-tau antibodies examined. The areas of distribution of granule-containing neurons detected by the Gallyas-Braak method appeared to overlap with the reported main sites of subcortical distribution of neurofibrillary tangles. They, however, were not observed in the cerebral cortex, including the hippocampal region. The Alz-50 immunoreactive granules showed ultrastructural features similar to those of lysosomes or lipofuscin. These findings suggest that denatured tau might be degraded in lysosomes.


Synthesis and characterization of a new Positron emission tomography probe for orexin 2 receptors neuroimaging.
The orexin receptors (OXRs) have been involved in multiple physiological and neuropsychiatric functions. Identification of PET imaging probes specifically targeting OXRs enables us to better understand the OX system. Seltorexant (JNJ-42847922) is a potent OX2R antagonist with the potential to be an OX2R PET imaging probe. Here, we describe the synthesis and characterization of [18F]Seltorexant as an OX2R PET probe. The ex vivo autoradiography studies indicated the good binding specificity of [18F]Seltorexant. In vivo PET imaging of [18F]Seltorexant in rodents showed suitable BBB penetration with the highest brain uptake of %ID/cc = 3.4 at 2 min post-injection in mice. The regional brain biodistribution analysis and blocking studies showed that [18F]Seltorexant had good binding selectivity and specificity. However, pretreatment with unlabelled Seltorexant and P-gp competitor CsA observed significantly increased brain uptake of [18F]Seltorexant, indicating [18F]Seltorexant could interact P-gp at the blood-brain barrier. Our findings demonstrated that [18F]Seltorexant is a potential brain OX2R PET imaging probe, which paves the way for new OX2R PET probes development and OX system investigation.


DNA methylation markers in peripheral blood for psoriatic arthritis.
The clinical manifestations of psoriatic arthritis (PsA) are highly heterogeneous and no reliable diagnostic biomarkers exist. We explored the role of DNA methylation CpG markers in the diagnosis of PsA. DNA methylation array was used to screen for differentially methylated sites (DMSs) in the discovery phase (PsA, n = 25; healthy controls [HCs], n = 19; psoriasis vulgaris [PsV], n = 20). In the validation phase, pyrosequencing was used to identify the DMSs in an expanded cohort (PsA, n = 60; HCs, n = 91; PsV, n = 48; rheumatoid arthritis [RA], n = 60). Logistic regression prediction models were established based on the identified DMSs for the diagnosis of PsA. A total of 17 DMSs differentiating PsA and HCs as well as 11 DMSs differentiating PsA and PsV were screened in the discovery phase. A total of six DMSs (chr14: cg07940072, chr14: 38061320, chr9: cg15734589, chr6: cg12800266, chr3: cg12992827, chr6: cg24500972) differentiating PsA and HCs and two DMSs (chr12: cg16459382, chr2: cg16348668) differentiating PsA and PsV were identified using pyrosequencing. Three logistic regression prediction models were established based on the identified DMSs, which distinguished PsA, RA, PsV, and HCs (P < 0.001). The models performed well in differentiating PsA from HCs, RA, and PsV (AUC: 0.858, 0.851, and 0.976, respectively). The models based on methylated CpG sites are useful for distinguishing patients with PsA from HCs and those with RA or PsV and are a highly sensitive and specific diagnostic biomarker for PsA.


Pelvifemoral rhythm during unilateral hip flexion in standing.
To quantify pelvifemoral rhythm during unilateral hip flexion in standing subjects, under different degrees of resistance. Descriptive and correlational. Pelvic rotation contributes to the change in hip flexion angle while individuals are supine. Degree of contribution in standing has not been described. Pelvic and femoral motions, relative to vertical, were documented while 14 subjects performed three trials of standing unilateral hip flexion, both with and without a 4.53 kg ankle weight. Pelvic and thigh segments were marked with reflective spheres, and motion was captured by video analysis. Pelvic rotation contributed 18.1% of the change in hip flexion angle. Pelvic rotation occurred throughout the movement. Neither presence of external resistance nor direction of movement significantly changed this contribution. A strong relationship exists between pelvic rotation and hip flexion in standing subjects. Hip flexion, while standing, is a common element of many functional activities. Knowledge of the pelvic contribution increases our understanding of hip and lumbar spine motion during standing activities.


Disagreement between patient- and physician-reported outcomes on symptomatic adverse events as poor prognosis in patients treated with first-line cetuximab plus chemotherapy for unresectable metastatic colorectal cancer: Results of Phase II QUACK trial.
The status and prognostic value of the disagreement between physician and patient assessments of symptomatic adverse events (AEs) remain unclear for patients with metastatic colorectal cancer treated with first-line cetuximab plus chemotherapy. Paired data on patient-reported outcomes using the EORTC QLQ-C30 and physician-reported outcomes using the NCI-CTCAE for eight symptomatic AEs (fatigue, pain, insomnia, dyspnea, constipation, appetite loss, nausea/vomiting, and diarrhea) were collected from a prospective trial assessing the relationships between treatment efficacy, AEs, and quality of life. The overall agreement rates between patient and physician reporting at 4 weeks ranged from 40.2% to 76.5% for 129 patients. The level of agreement based on Cohen's κ statistics was slight to poor for dyspnea, pain, fatigue, and insomnia, while it was moderate to fair for the remaining AEs. No clinicopathological characteristics of disagreement were found. The underreporting by physicians ranged from 12.5% (nausea/vomiting) to 56.7% (fatigue). The 2-year overall survival (OS) rate was more favorable for patients with high agreement than for those with low agreement (71.2% vs. 46.5%, p = .016), and the agreement status was an independent factor of OS (HR, 2.31; 95% CI, 1.13-4.71; p = .022). For patients who were reported as asymptomatic by the physician, the presence of patient-reported symptoms resulted in a trend toward poor prognostic outcomes for appetite loss, dyspnea, diarrhea, and constipation. These findings provide the clinical importance of the monitoring of patient-reported symptoms that can be complementary to physician-reported data to ensure more accurate clinical outcomes.


Using Graph-Based Signatures to Guide Rational Antibody Engineering.
Antibodies are essential experimental and diagnostic tools and as biotherapeutics have significantly advanced our ability to treat a range of diseases. With recent innovations in computational tools to guide protein engineering, we can now rationally design better antibodies with improved efficacy, stability, and pharmacokinetics. Here, we describe the use of the mCSM web-based in silico suite, which uses graph-based signatures to rapidly identify the structural and functional consequences of mutations, to guide rational antibody engineering to improve stability, affinity, and specificity.


CKD Progression Prediction in a Diverse US Population: A Machine-Learning Model.
Chronic kidney disease (CKD) is a major cause of morbidity and mortality. To date, there are no widely used machine-learning models that can predict progressive CKD across the entire disease spectrum, including the earliest stages. The objective of this study was to use readily available demographic and laboratory data from Sonic Healthcare USA laboratories to train and test the performance of machine learning-based predictive risk models for CKD progression. Retrospective observational study. The study population was composed of deidentified laboratory information services data procured from a large US outpatient laboratory network. The retrospective data set included 110,264 adult patients over a 5-year period with initial estimated glomerular filtration rate (eGFR) values between 15-89 mL/min/1.73 m2. Patient demographic and laboratory characteristics. Accelerated (ie, >30%) eGFR decline associated with CKD progression within 5 years. Machine-learning models were developed using random forest survival methods, with laboratory-based risk factors analyzed as potential predictors of significant eGFR decline. The 7-variable risk classifier model accurately predicted an eGFR decline of >30% within 5 years and achieved an area under the curve receiver-operator characteristic of 0.85. The most important predictor of progressive decline in kidney function was the eGFR slope. Other key contributors to the model included initial eGFR, urine albumin-creatinine ratio, serum albumin (initial and slope), age, and sex. The cohort study did not evaluate the role of clinical variables (eg, blood pressure) on the performance of the model. Our progressive CKD classifier accurately predicts significant eGFR decline in patients with early, mid, and advanced disease using readily obtainable laboratory data. Although prospective studies are warranted, our results support the clinical utility of the model to improve timely recognition and optimal management for patients at risk for CKD progression. Defined by a significant decrease in estimated glomerular filtration rate (eGFR), chronic kidney disease (CKD) progression is strongly associated with kidney failure. However, to date, there are no broadly used resources that can predict this clinically significant event. Using machine-learning techniques on a diverse US population, this cohort study aimed to address this deficiency and found that a 5-year risk prediction model for CKD progression was accurate. The most important predictor of progressive decline in kidney function was the eGFR slope, followed by the urine albumin-creatinine ratio and serum albumin slope. Although further study is warranted, the results showed that a machine-learning model using readily obtainable laboratory information accurately predicts CKD progression, which may inform clinical diagnosis and management for this at-risk population.


Comparative Whole-Transcriptome Profiling of Liver Tissue from Wistar Rats Fed with Diets Containing Different Amounts of Fat, Fructose, and Cholesterol.
Differential expression of 30,003 genes was studied in the liver of female Wistar rats fed with isocaloric diets with the excess of fat, fructose, or cholesterol, or their combinations for 62 days using the method of whole-transcriptome profiling on a microchip. Relative mRNA expression levels of the Asah2, Crot, Crtc2, Fmo3, GSTA2, LOC1009122026, LOC102551184, NpY, NqO1, Prom1, Retsat, RGD1305464, Tmem104, and Whsc1 genes were also determined by RT-qPCR. All the tested diets affected differently the key metabolic pathways (KEGGs). Significant changes in the expression of steroid metabolism gene were observed in the liver of animals fed with the tested diets (except the high-fat high fructose diet). Both high-fat and high-fructose diets caused a significant decrease in the expression of squalene synthase (FDFT1 gene) responsible for the initial stage of cholesterol synthesis. On the contrary, in animals fed with the high-cholesterol diet (0.5% cholesterol), expression of the FDFT1 gene did not differ from the control group; however, these animals were characterized by changes in the expression of glucose and glycogen synthesis genes, which could lead to the suppression of glycogen synthesis and gluconeogenesis. At the same time, this group demonstrated different liver tissue morphology in comparison with the animals fed with the high-fructose high-fat diet, manifested as the presence of lipid vacuoles of a smaller size in hepatocytes. The high-fructose and high-fructose high-fat diets affected the metabolic pathways associated with intracellular protein catabolism (endocytosis, phagocytosis, proteasomal degradation, protein processing in the endoplasmic reticulum), tight junctions and intercellular contacts, adhesion molecules, and intracellular RNA transport. Rats fed with the high-fructose high-fat or high-cholesterol diets demonstrated consistent changes in the expression of the Crot, Prom1, and RGD1305464 genes, which reflected a coordinated shift in the regulation of lipid and carbohydrate metabolisms.


Simple association analysis combining data from trios/sibships and unrelated controls.
We consider genetic association analysis that combines data from case-parent trios/sibships and unrelated controls. A general and simple methodology is proposed, using a weighted least-squares approach to combine separate information from the case-parent/case-sibling analysis and the case-unrelated control analysis. The new proposal improves over the existing methods in that it requires no assumptions and estimation on the mating-type distribution. Simulation results show that the new method competes well with the likelihood-based method when the latter is applicable, and achieves substantial power gains over separate analyses in general. Therefore, the proposed combined association analysis can enjoy wide applications, including the multiallele/locus, haplotype, and genome-wide association studies.


Chlorhexidine exposure of clinical Klebsiella pneumoniae strains leads to acquired resistance to this disinfectant and to colistin.
Chlorhexidine is widely used as a disinfectant in hospitals, which may impose a selective pressure on bacteria. This study aimed to determine whether continuous exposure to chlorhexidine could lead to adaptive resistance and cross-resistance as well as investigating potential resistance mechanisms. Three clinical Klebsiella pneumoniae strains susceptible to conventional antimicrobials were selected and were continuously cultured in broth with gradually increasing concentrations of chlorhexidine. Antimicrobial susceptibility was determined. Mechanisms of acquired resistance to chlorhexidine and colistin were analysed by PCR and reverse transcription quantitative PCR (RT-qPCR). Furthermore, fitness was assessed through growth curve assays. Increased resistance to chlorhexidine and colistin was observed in all strains. Expression of the cepA gene was upregulated in the adapted strains, suggesting that hyperexpression of CepA was probably the main mechanism of adaptive resistance to chlorhexidine. The amino acid substitutions Leu82Arg and Arg256Gly in PmrB were detected in all of the adapted strains, whilst Leu344Pro was only identified in one adapted strain, indicating that the PmrB substitution was responsible for the cross-resistant phenotype. Moreover, chlorhexidine adaptation might have an effect on bacterial growth.


National intercomparison program for radiopharmaceutical activity measurements.
Since 1998, the National Laboratory for Ionizing Radiation Metrology has lead an intercomparison program for activity measurements of radiopharmaceuticals administered to patients in the Nuclear Medicine Services (NMS) with the purpose of promoting the quality control. In this work, intercomparison results with the radionuclides 67Ga, 123I, 99mTc and 201Tl from the beginning of the program to the present will be presented establishing as a consequence, the radionuclide calibrators' traceability used by the participants. The analysis of the results showed that 68% were within the limits of +/-10% required by the Brazilian Regulatory Authority that demands this accuracy band for the radiopharmaceutical measurements in the NMS. The results also demonstrate an improvement in the measurement quality that this program has provided since its beginning.


Ocular involvement in primary antiphospholipid syndrome. Ocular involvement in primary APS.
The purpose of this study is to evaluate the ocular findings in patients with the primary antiphospholipid syndrome (APS). Twenty-two patients (44 eyes) with primary APS (17 women, 5 men) were examined. All patients were younger than 50 years (median age; 37.5 years). In 18 patients, fundus flourescein angiography was performed in addition to the ophthalmologic examination. Sixteen patients (72.7%) described visual symptoms. Anterior segment was normal in 19 patients (86.4%). Posterior segment abnormalities were observed in 15 patients (68.2%). Venous dilatation and tortuosity were the most common ocular findings. Retinal vascular occlusive disease was detected in 5 patients (22.7%). Flourescein angiography abnormalities were noted in 14 of the 18 patients (77.8%). The most common angiographic finding was pigment epithelial window defects. Our results indicate that posterior eye segment involvement is relatively common in the primary APS. It also seems that the screening for APS is important in young patients with retinal vascular occlusion, especially in those without conventional risk factors.


Simple light guide for measuring irradiance in an aqueous oxygen electrode chamber.
The light-dependent reactions of photosynthesis are often measured with Clark-type oxygen electrodes yet the irradiance level inside aqueous oxygen electrode reaction vessels is seldom reported due to the difficulty of measuring light inside a small volume chamber. We describe a simple light guide terminating in a 90 degrees prism that can be inserted into a reaction vessel. Incoming irradiation is directed to a commercially available quantum sensor positioned at the other end of the light guide. Both materials for and construction of the device are inexpensive.


Effect of in vitro gastrointestinal digestion on the antibacterial activity of bioactive dairy formulas supplemented with lactoferrin against Cronobacter sakazakii.
Milk is a source of proteins with high nutritional value and relevant biological activities. Bioactive milk proteins, like lactoferrin, are important for newborn development and can also be used as ingredients in functional products to improve health. Lactoferrin is essential in infant's diet, since protects against infections and promotes immune system maturation. Bovine lactoferrin is used to supplement formula milk in order to strengthen baby's defences against some pathogenic bacteria. Thus, lactoferrin supplemented formula can be a barrier against emergent pathogens, such as Cronobacter sakazakii, which has caused great concern in the last few years. Milk proteins generate bioactive peptides in the digestion process, and it is known that industrial processing can modify their susceptibility to digestion. Treatments such as heating have been shown to denature whey proteins and make them more easily digestible. Therefore, the aim of this study was to analyze the effect of technological treatments and gastrointestinal digestion on the antibacterial activity against C. sakazakii of proteins present in dairy formulas supplemented with lactoferrin. Commercial bovine lactoferrin has been shown to have antibacterial activity against C. sakazakii, both in the native state and after static in vitro gastrointestinal digestion. In addition, the digests obtained from dairy formulas subjected to technological treatments, either homogenization or pasteurization, have higher antibacterial activity than non-treated formulas. The release of low molecular weight peptides during the in vitro gastric digestion is probably the cause that would explain the enhanced antibacterial activity of the digested dairy formulas.


A novel PDZ protein regulates the activity of guanylyl cyclase C, the heat-stable enterotoxin receptor.
Secretory diarrhea is the leading cause of infectious diarrhea in humans. Secretory diarrhea may be caused by binding of heat-stable enterotoxins to the intestinal receptor guanylyl cyclase C (GCC). Activation of GCC catalyzes the formation of cGMP, initiating a signaling cascade that opens the cystic fibrosis transmembrane conductance regulator chloride channel at the apical cell surface. To identify proteins that regulate the trafficking or function of GCC, we used the unique COOH terminus of GCC as the "bait" to screen a human intestinal yeast two-hybrid library. We identified a novel protein, IKEPP (intestinal and kidney-enriched PDZ protein) that associates with the COOH terminus of GCC in biochemical assays and by co-immunoprecipitation. IKEPP is expressed in the intestinal epithelium, where it is preferentially accumulated at the apical surface. The GCC-IKEPP interaction is not required for the efficient targeting of GCC to the apical cell surface. Rather, the association with IKEPP significantly inhibits heat-stable enterotoxin-mediated activation of GCC. Our findings are the first to identify a regulatory protein that associates with GCC to modulate the catalytic activity of the enzyme and provides new insights in mechanisms that regulate GCC activity in response to bacterial toxin.


Separation of stereoisomers of some terpene derivatives by capillary gas chromatography-mass spectrometry and high-performance liquid chromatography using beta-cyclodextrin derivative columns.
Gas chromatographic separations of the stereoisomers of menthol derivatives, important intermediates in the synthesis of physiologically active natural products, were carried out on several substituted beta-cyclodextrin (CD) columns, including per-O-methyl-beta-cyclodextrin (PME-beta-CD), heptakis(2,3-di-O-acetyl-6-tert-butyldimethylsilyl)-beta-CD (DIAC-6-TBDS-beta-CD), and heptakis(2,3-di-O-methyl-6-tert-butyldimethylsilyl)-beta-CD (DIME-6-TBDS-beta-CD) as chiral stationary phases (CSPs). With the DIME-6-TBDS-beta-CD column, a separation of the Z- and E-isomers of methylidenementhol was accomplished; no separation was achieved with the other columns. The stereoisomers of methylidenementhol and the corresponding tert-butyldimethylsilyl (TBS) ether were separated on both the beta-CD and the heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin (TME-beta-CD) columns by high-performance liquid chromatography (HPLC) with a mobile phase involving acetonitrile and H(2)O. For the separation of the Z- and E-isomers of methylidenementhol, the TME-beta-CD column was superior. In contrast, the beta-CD column was preferable in the case of the corresponding TBS ether.


CT-findings in cystic meningiomas.
Cystic meningiomas are rare, about 10% occurring in children. Only 61 cases have been reported in the literature. Based on 6 of our own cases and the reports in the literature typical CT-findings are described and differentiating features from other cystic mass lesions discussed. If a tumour on CT shows a cystic component, is smoothly outlined, enhances markedly and is attached to the dura, then a preoperative diagnosis of cystic meningioma should be taken into account. This holds especially true if the patient is a child.


Medicare chronic care improvement program puts the spotlight on case management.
A 3-year initiative to provide chronic care services for Medicare fee-for-service (FFS) beneficiaries with diabetes or congestive heart failure is putting the spotlight on how case management teamed with disease management can improve clinical outcomes, generate cost savings, and improve patient and provider satisfaction.


Reverse shoulder arthroplasty after failed pectoralis major tendon transfer with a minimum follow-up of 5 years. A case series.
Treatment strategies after failed pectoralis major tendon transfer (PMTT) are scarce in literature and no information is available for patients treated with reverse shoulder arthroplasty (RSA) for failed PMTT. We performed a retrospective outcome study of all patients who underwent revision with RSA after failed PMTT at our institution. From 1999 to 2009 we included 8 patients (8 shoulders). The minimum follow-up was 5 years with a mean follow-up time of 97 months (range, 64-134). Clinical and radiological evaluation comprised range of motion, Constant Murley score (CMS), Simple Shoulder Test (SST) as well as standard X-rays in 2 planes. Pain was measured using VAS pain scale. The patients were asked to rate their satisfaction at final follow-up. All outcome measures improved significantly post-surgical compared to pre-surgical, including the CMS (17.8-62.8), SST (1.8-7.3) and VAS (7.1-1). Active abduction and anterior flexion improved significantly (p < .001) from 65.6° to 125° and from 62.5° to 136.3°, respectively. There were 2 complications. One patient sustained transient musculocutaneous nerve palsy that resolved completely after 5 months and one patient sustained postoperative hematoma and had to be revised 4 days after surgery. 4 (50%) patients rated their results as excellent and 4 (50%) rated them as good. RSA is a good option for treating patients after failed pectoralis major tendon transfer. After a minimum follow-up of 5 years, functional outcome is good and patient satisfaction is high. Level IV, Case series, Treatment study.


Interactions between light and growing season temperatures on, growth and development and gas exchange of Semillon (Vitis vinifera L.) vines grown in an irrigated vineyard.
High-light intensities and temperatures of the warm climate regions of Australia and elsewhere have a major effect on the growth and development of grapevines (Vitis vinifera L.). The objective of this research was to assess interactions between the light and seasonal temperatures by shading some vines and comparing these with vines exposed to high-light intensities. Canopy temperatures were monitored using infrared radiometers and budbreak, phenology, growth, yield, berry ripening and gas exchange determined over three growing seasons. Results showed canopies were generally about 4 °C cooler than air and shading extended this cooling. Irradiance, irrespective of seasonal temperatures, had no effect on time of budbreak, shoot phenology, stem growth, yield and bunch fresh weights while bunch and leaf dry weights were reduced in low-light. Bunch ripening was initially delayed by low-light but thereafter the ripening process was highly temperature-dependent. Rates increased linearly with increasing temperature in both low and high-light and were optimal at about 35 °C. Maximum photosynthetic capacity was impaired by low irradiance, in accordance with shade leaf attributes, and attributable to stomatal closure. No effects of the low photosynthetic capacity apparently carried-over to sugar accumulation, consistent with the strong sink capacity of bunches.


Great debates in trauma biomechanics.
At the 2021 annual meeting of the Orthopaedic Trauma Association, the Basic Science Focus Forum hosted its first ever debate-style symposium focused on biomechanics and fracture repair. The 3 subjects of debate were "Mechanics versus Biology-Which is 'More Important' to Consider?" "Locked Plate versus Forward Dynamization versus Reverse Dynamization-Which Way Should I Go?" and "Sawbones versus Cadaver Models-What Should I Believe Most?" These debates were held because fracture healing is a highly organized synergistic response between biological factors and the local mechanical environment. Multiple studies have demonstrated that both factors play roles in governing bone healing responses, and the causal relationships between the 2 remain unclear. The lack of clarity in this space has led to a spectrum of research with the common goal of helping surgeons make good decisions. Before reading further, the reader should understand that the questions posed in the debate titles are unanswerable and might represent a false choice. Instead, the reader should appreciate that the debates were held to gain a more thorough understanding of these topics based on the current state of the art of experimental and clinical studies, by using an engaging and thought-provoking format.


Histopathological evaluation of anterior lamellar corneal tissue-on/-off storage conditions on DSAEK donor tissue after storage in organ culture.
To compare the quality of corneal endothelium of precut Descemet-stripping automated endothelial keratoplasty (DSAEK) tissue when transported with and without the anterior lamellar corneal tissue (ALCT) when organ-culture corneal storage methods are used. After microkeratome-assisted excision of anterior corneal lamella, five pairs of corneas (10 eyes) were stored either with the ALCT on the stroma (five eyes) or with ALCT off the stroma (five eyes) in organ-culture medium. Three pairs (six matched corneas) were left in the transport medium for 24 h prior to the microkeratome cut. Two pairs (four matched corneas) were left in the transport medium for 48 h prior to the microkeratome cut. All cuts were performed using a 300 (four eyes) or 350 (six eyes) microns head. A vital dye assay (alizarin red S and trypan blue) was used to identify devitalized and necrotic endothelial cells. In all matched cases, there was no difference between the endothelial cell appearance with or without the anterior corneal lamella. In all cases, there was no evidence for trypan blue stained cells beyond that normally seen on acceptable transplantable corneas and there was no evidence of loss of cells or any lifting of Descemet's membrane. There is no difference between the quality of the donor endothelial cell appearance with ALCT-on or -off when the donor cornea is stored using the organ-culture system of corneal storage. Organ-culture storage system is a safe and effective system in regard to preparation and transport of donor lenticules for DSAEK.


Cytotoxic Sesquiterpenoids from Atractylodes chinensis (DC.) Koidz.
Four new sesquiterpenoids named atrchiterpenes A-D (1-4), a new natural product (5), and twelve known compounds (6-17) were isolated from Atractylodes chinensis (DC.) Koidz. Compound 1 was a rare N-containing eudesmane-type sesquiterpenoid. Structure elucidation was performed by spectroscopic techniques, including 1D, 2D NMR spectra, and HR-ESI-MS. Compounds 6-11, 14, and 17 were reported from Atractylodes for the first time. All the isolated compounds were evaluated for cytotoxicity activity. Compound 16 showed moderate cytotoxicity against HepG2 cells with an IC50 value of 5.81±0.47.


Structure of the O-polysaccharide from the lipopolysaccharide of Providencia alcalifaciens O33.
Mild acid degradation of the lipopolysaccharide from Providencia alcalifaciens O33 resulted in an O-polysaccharide along with core and O-unit-bearing core oligosaccharides. Composition of the oligosaccharides was inferred by ESI mass spectrometry. Based on sugar and methylation analyses, Smith degradation and (1)H and (13)C NMR spectroscopy data, the following structure of the tetrasaccharide O-unit of the O-polysaccharide was established: Another O-polysaccharide structure has been reported earlier for Providencia stuartii О33 but later found to belong to a P. stuartii О52 strain.


Nearest-neighbour graphical analysis of spatial pattern and a test for competition in populations of singing crickets (Teleogryllus commodus).
The most crowded individuals in a population often compete for space and develop a regular pattern of spacing. Such regularity is not normally recognized because it occurs within high-density regions of a populated area showing overall aggregation. Thus competition for space, as reflected by spatial pattern, often goes undetected when standard tests for spatial randomness are used. The test described in this paper makes use of truncated samples of nearest-neighbour distances arranged in ascending order, so that only the pattern of spacing of the most crowded individuals is analysed. This is the basis of Pielou's test (1962, 1977) for spatial competition. An advantage of the approach described is that the density of the most crowded individuals is determined graphically. The graphical method also provides a means for choosing an appropriate, non-arbitrary, truncation point for the test. The test was applied to two samples of singing crickets to demonstrate the procedure. The biological significance of the spacing patterns identified is discussed.


Functionality Assessment of Patients With Cleft Hands.
Ectrodactyly, commonly referred to as cleft hand, is a rare pathology characterized by a deficiency and/or complete absence of the central ray in each hand. In order to customize treatment and improve the patient's quality of life, a more detailed functional evaluation is required. Although several studies evaluate functionality in different types of cleft hands, there are only a few studies that show self-reported evaluations. The objective of this study is to assess the hand function of cleft hand patients. An observational retrospective study was performed on 12 cleft hand patients who were treated between 2008 and 2018. There were 8 male patients and 4 female patients. Patients were divided into 2 groups according to their ages: (Group 1) 6 patients between 1 and 7 years of age, and (Group 2) 6 patients between 8 and 18 years of age, respectively. Each group was sub-stratified into 5 subgroups according to the classification system created by Manske and Halikis. Regardless of age, intragroup hand type comparisons within Groups 1 and 2 did not demonstrate statistically significant differences (P > 0.05) between hand outcomes according to Manske and Halikis classification. Comparison between cleft hand patients and their age matched controls demonstrated statistically significant differences (P < 0.05), as the patients in the control group had higher outcome scores. Regardless of cleft hand type and patient age, patients with cleft hands experience impaired hand function and present lower outcome scores in comparison to their age matched controls.


Urethral pressure variations in healthy females during rest and sleep.
Recent studies have shown the occurrence of rhythmic urethral pressure variations (UPV) in the normal female urethra in the awake state. The nature of these variations still needs further elucidation. In this study we investigated the changes in urethral pressure (UP) and in UPV as they occur in healthy females awake and asleep. UPV, intravesical-(IVP) and rectal pressure (RP), together with anal electromyography (EMG) and electroencephalography (EEG) were measured in 10 healthy fertile female volunteers. The registration was done in the awake state, during falling asleep and during sleep. During falling asleep the average urethral pressure as well as the UPV amplitude and frequencies decreased and remained at a lower stable level during sleep. Urethral pressure and its variations seemed to be interrelated since the UPV frequency and amplitude decreased when the average UP decreased. From this study it appears that urethral pressure is influenced by extrinsic factors such as the waking, resting and sleeping states. Although some extrinsic modulation of urethral pressure variations occurs, their persistance in sleep suggests their local nature.


Simultaneous production of immunoaffinity membranes.
We simultaneously separated antibodies for transferrin, the third component of complement (C3), haptoglobin and transthyretin by multi-sample non-denaturing two-dimensional electrophoresis (2-DE), transferred them to a polyvinylidene difluoride (PVDF) membrane and then stained them using direct blue 71 to obtain membrane-immobilized antibodies. The antigens, transferrin, C3, haptoglobin and transthyretin were specifically bound to the membrane-immobilized antibodies and were eluted only after rinsing the membrane with acid solution. The antigens specifically bound to the membrane-immobilized antibodies were separated by SDS-PAGE and identified by peptide mass fingerprinting using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Furthermore, transferrin and transthyretin were trapped and eluted by each membrane-immobilized antibody and detected by MALDI-TOF MS directly without separations. Using membrane-immobilized anti-transferrin antibody, transferrin in flowing blood was directly trapped and analyzed. The results indicated that membrane-immobilized antibodies are simultaneously produced, and that the immunoaffinity membranes can capture specific substances in flowing fluids.


Anodic concentration loss and impedance characteristics in rotating disk electrode microbial fuel cells.
A rotating disk electrode (RDE) was used to investigate the concentration loss and impedance characteristics of anodic biofilms in microbial fuel cells (MFCs). Amperometric time-current analysis revealed that at the rotation rate of 480 rpm, a maximum current density of 168 µA cm(-2) can be achieved, which was 22.2 % higher than when there was no rotation. Linear sweep voltammetry and electrochemical impedance spectroscopy tests showed that when the anodic potential was set to -300 mV vs. Ag/AgCl reference, the power densities could increase by 59.0 %, reaching 1385 mW m(-2), the anodic resistance could reduce by 19 %, and the anodic capacitance could increase by 36 %. These results concur with a more than 85 % decrease of the diffusion layer thickness. Data indicated that concentration loss, diffusion layer thickness, and the mixing velocity play important roles in anodic resistance reduction and power output of MFCs. These findings could be helpful to the design of future industrial-scale MFCs with mixed bacteria biofilms.


Data-driven comorbidity analysis of 100 common disorders reveals patient subgroups with differing mortality risks and laboratory correlates.
The populational heterogeneity of a disease, in part due to comorbidity, poses several complexities. Individual comorbidity profiles, on the other hand, contain useful information to refine phenotyping, prognostication, and risk assessment, and they provide clues to underlying biology. Nevertheless, the spectrum and the implications of the diagnosis profiles remain largely uncharted. Here we mapped comorbidity patterns in 100 common diseases using 4-year retrospective data from 526,779 patients and developed an online tool to visualize the results. Our analysis exposed disease-specific patient subgroups with distinctive diagnosis patterns, survival functions, and laboratory correlates. Computational modeling and real-world data shed light on the structure, variation, and relevance of populational comorbidity patterns, paving the way for improved diagnostics, risk assessment, and individualization of care. Variation in outcomes and biological correlates of a disease emphasizes the importance of evaluating the generalizability of current treatment strategies, as well as considering the limitations that selective inclusion criteria pose on clinical trials.


Clinical evaluation of the CORDIS vascular access port systems: a multicenter study.
Two non-metallic vascular access port systems, the Multipurpose Access Port (MPAP) and Miniport, developed by CORDIS S.A., France, have been evaluated clinically in 78 cancer patients. During the investigational period covering a total experience of 369 treatment cycles and 1,370 infusion days, no cases of infection or septicemia were observed. Serious complications such as drug extravasation and catheter occlusion occurred, although the incidence was relatively low (+/- 1%) when compared with the number of treatment courses (cycles), but in relation to the number of patients included in this study, the procedure-related complication rate was 17.5% for the MPAP and 15.8% for the Miniport. Procedure-related complications can be avoided by proper handling and use of suitable drug combinations to minimize crystallization reactions within the port-catheter systems. The final complication rate (total minus procedure-related) in terms of termination of treatment, i.e. explantation of the port-catheter system was 12.1% for the MPAP and 12.5% for the MINIPORT, which generally confirms the results of other groups. More than 87% of both port-catheter systems were still functional at the end of evaluation.


Changes in adenosine deaminase activity in ageing cultured human cells and the role of zinc.
The level of adenosine deaminase (ADA; EC 126.96.36.199) was estimated at different passages in six confluent fibroblast cultures established from forearm skin biopsies of healthy adult normal volunteers. After determination of the zinc concentration in standard growth medium, ADA activity was estimated at different passages of subculture in media with different zinc concentrations. The results indicated that the specific activity of ADA in control confluent skin fibroblast cultures (passage 2) cultivated in standard growth medium containing 15.4 microM zinc (similar to that present in normal human plasma) was equal to 226.6+/-19.64 micromol min(-1) mg(-1) protein. The results showed that there were no significant changes in ADA specific activity in any of the control cultures as the zinc concentration of the medium was increased. To characterize the passage of subculture at which fibroblasts enter the ageing phase, three marker enzymes were assayed namely, phosphofructokinase, lactate dehydrogenase and glycogen phosphorylase. The result showed that the cells enter the ageing phase at passage 20 and beyond. Further investigation showed that ADA activity of serially subcultured confluent cultures cultivated in standard growth medium significantly dropped at passages 20, 25 and 30. ADA activity however was not significantly altered in cells at passage 2, 10 and 15 cultivated in standard growth medium and in the presence of higher zinc levels (23.1, 34.6, 53.8 and 73.1 microM). Furthermore there was significant lowering of ADA activities in cells at passages 20, 25 and 30 when cells were cultured in the presence of 15.4, 23.1 and 34.6 microM zinc. Such lowered activities of ADA were restored to normal when the cells were cultured in the presence of higher zinc concentration equal to 53.8 and 73.1 microM. From the results we concluded that it is possible to restore ADA activity in aged skin fibroblasts to normal levels by raising the zinc concentration in the culture medium to four or five times the control normal plasma zinc level.


The invasion and growth of Babesia bovis in tick tissue culture.
Erythrocytic forms of Babesia bovis inoculated into cell cultures of the tick Boophilus microplus invaded the tick cells and showed multiplication for up to 48 h after inoculation.


Human herpesvirus 6 infections after bone marrow transplantation: clinical and virologic manifestations.
Human herpesvirus 6 (HHV-6) DNA levels in peripheral blood mononuclear cells were prospectively evaluated in 20 cytomegalovirus-seronegative allogeneic marrow transplant patients and in 10 healthy control subjects. Blood and saliva specimens obtained weekly for 3 months after transplant were evaluated by quantitative HHV-6 polymerase chain reaction. One of 20 patients experienced primary HHV-6 infection after marrow transplant (seroconversion, HHV-6 viremia, skin rash); 18 of 20 had increased peripheral blood mononuclear cell HHV-6 DNA levels consistent with asymptomatic reactivations, and 1 patient experienced a reactivation-associated skin rash. Genotyping revealed HHV-6 variant B DNA in all cases. Therapy with acyclovir or intravenous immunoglobulin was not correlated with lower HHV-6 DNA levels. Thus, asymptomatic HHV-6 reactivations appear to be common following allogeneic marrow transplantation. Among HHV-6-seronegative and viral DNA-negative patients, primary HHV-6 infection can ensue in association with self-limited clinical symptoms, including diffuse maculopapular rash.


A Unique Immunofluorescence Protocol to Detect Protein Expression in Vascular Tissues: Tacking a Long Standing Pathological Hitch.
Autofluorescence induced interference is one of the major drawbacks in immunofluorescence analysis of formalin-fixed paraffin-embedded tissues, as it decreases the signal-to-noise ratio of specific labeling. Apart from aldehyde-fixation induced artifacts; collagen and elastin, red blood cells and endogenous fluorescent pigment lipofuscin are prime sources of autofluorescence in vascular and aging tissues. We describe herein, an optimized indirect-immunofluorescence method for archival formalin-fixed paraffin-embedded tissues tissues and cryo sections, using a combination of 3-reagents in a specific order, to achieve optimal fluorescence signals and imaging. Human telomerase reverse transcriptase, a protein implicated as a proliferation marker, was chosen relevant to its expression in solid tumors along with 3 other intracellular proteins exhibiting nuclear and/or cytoplasmic expression. Staining was performed on 10 glioma tissue sections along with 5 of their cryo sections, 5 sections each of hepatocellular, lung, papillary-thyroid and renal cell carcinoma, with 10 non-malignant brain tissue samples serving as control. Specimens were imaged using epifluorescence microscopy, followed by software-based quantification of fluorescence signals for statistical analysis and validation. We observed that the combined application of sodium-borohydride followed by crystal violet before antigen retrieval and a Sudan black B treatment after secondary antibody application proved to be most efficacious for masking autofluorescence/non-specific background in vascular tissues. This unique trio-methodology provides quantifiable observations with maximized fluorescence signal intensity of the target protein for longer retention time of the signal even after prolonged storage. The results can be extrapolated to other human tissues for different protein targets.


Young's procedure for the treatment of valgus flatfoot deformity caused by a posterior tibial tendon dysfunction, stage II.
Young's procedure contains an action mechanism that works better than other techniques on the pathophysiology of FFD. It respects the anatomy and biomechanics of the foot to reach the necessary muscular balance. The benefits of this technique include that the ATT is not detached, so its function mechanism is still active; the new trajectory of the ATT provides a powerful sling function at the level of the navicular; and the horizontal trajectory of the ATT and the osteoperiosteal flaps constitute a powerful inner capsular–tendinous–ligamentous support. What is more, an insufficiency of the ATT is created, which results in a predominance of the peroneus lateral longus, that descends and prones the forefoot. Additional procedures, such as medial displacement calcaneal osteotomy, should be considered to correct the entire deformity. The combination of these techniques do not sacrifice the joint mobility.


Fluorouracil for the treatment of massive periretinal proliferation.
A single intravitreal injection of fluorouracil was effective in the treatment of an experimental model of massive periretinal proliferation. When given with an intravitreal injection of 250,000 heterologous fibroblasts, fluorouracil decreased the rate of tractional retinal detachment from 36.8% in controls (seven of 19 eyes) to 5.2% in treated animals (one of 19 eyes) at one week, and from 73.6% in controls (14 of 19 eyes) to 31.5% in treated animals (six of 19 eyes) after four weeks (P less than .05). Intraocular neovascularization was reduced from 52.6% in controls (ten of 19 eyes) to 5.2% in treated animals (one of 19 eyes) after one week and 36.8% in controls (seven of 19 eyes) to 5.2% in treated animals (one of 19 eyes) after four weeks. When supplemented by repeated 10-mg subconjunctival injections of fluorouracil, or in combination with intravitreally administered indomethacin, this effect appeared to be enhanced. Intravitreal and subconjunctival injections of fluorouracil were well tolerated and may prove to be of significant value in the treatment of human disease.


Seasonal variations and sources study by way of back trajectories and ANOVA for ambient air pollutants (particulates and metallic elements) within a mixed area at Longjing, central Taiwan: 1-year observation.
This study measured the concentrations of particulates and metallic elements in ambient air by using PS-1 sampler (TSP) at Longjing area. And this study focuses on the collection of ambient air particulates, metallic elements, particulate-bound mercury Hg(p), concentrations. In addition, the sources of ambient pollutants by way of back trajectory analysis are found. Moreover, test mean concentration variance differences for metallic elements (PM, Hg(p), Mn, Fe, Zn, Cr, Cu, and Pb) among the four seasons (spring, summer, autumn and winter) through ANOVA are calculated. The result indicates that the average highest particulate concentration occurred in winter season, and the order was winter > spring > autumn > summer, and the mostly highest average metallic element (Mn, Fe, Zn, Cr, Cu, Pb) concentrations occurred in autumn. Moreover, the mostly average lowest metallic element concentrations occurred in summer. In addition, the above results of backward trajectories that the major particulate pollutants parcel mainly come from northeastern Taiwan. Moreover, when comparing the results of the first half year to that of the second half year, the they indicated that all metallic elements displayed significant differences in concentrations except those of Hg(p), Mn, Fe, Zn. Finally, metallic element Hg(p) is the only one which showed no significant concentration difference from either seasonal variations or half-year observations.


Transcriptome analysis of air-breathing land slug, Incilaria fruhstorferi reveals functional insights into growth, immunity, and reproduction.
Incilaria (= Meghimatium) fruhstorferi is an air-breathing land slug found in restricted habitats of Japan, Taiwan and selected provinces of South Korea (Jeju, Chuncheon, Busan, and Deokjeokdo). The species is on a decline due to depletion of forest cover, predation by natural enemies, and collection. To facilitate the conservation of the species, it is important to decide on a number of traits related to growth, immunity and reproduction addressing fitness advantage of the species. The visceral mass transcriptome of I. fruhstorferi was enabled using the Illumina HiSeq 4000 sequencing platform. According to BUSCO (Benchmarking Universal Single-Copy Orthologs) method, the transcriptome was considered complete with 91.8% of ortholog genes present (Single: 70.7%; Duplicated: 21.1%). A total of 96.79% of the raw read sequences were processed as clean reads. TransDecoder identified 197,271 contigs that contained candidate-coding regions. Of a total of 50,230 unigenes, 34,470 (68.62% of the total unigenes) annotated to homologous proteins in the Protostome database (PANM-DB). The GO term and KEGG pathway analysis indicated genes involved in metabolism, phosphatidylinositol signalling system, aminobenzoate degradation, and T-cell receptor signalling pathway. Many genes associated with molluscan innate immunity were categorized under pathogen recognition receptor, TLR signalling pathway, MyD88 dependent pathway, endogenous ligands, immune effectors, antimicrobial peptides, apoptosis, and adaptation-related. The reproduction-associated unigenes showed homology to protein fem-1, spermatogenesis-associated protein, sperm associated antigen, and testis expressed sequences, among others. In addition, we identified key growth-related genes categorized under somatotrophic axis, muscle growth, chitinases and collagens. A total of 4822 Simple Sequence Repeats (SSRs) were also identified from the unigene sequences of I. fruhstorferi. This is the first available genomic information for non-model land slug, I. fruhstorferi focusing on genes related to growth, immunity, and reproduction, with additional focus on microsatellites and repeating elements. The transcriptome provides access to greater number of traits of unknown relevance in the species that could be exploited for in-depth analyses of evolutionary plasticity and making informed choices during conservation planning. This would be appropriate for understanding the dynamics of the species on a priority basis considering the ecological, health, and social benefits.


Umbilical cord blood gas assessment of twins.
Umbilical cord arterial and venous blood gas values were compared in 63 twin pairs, of which 57 pairs had birth weights of 1500 g or more each. Small differences between the first and second twins existed for PO2, PCO2, and pH. However, bicarbonate values did not differ significantly. These cord gas differences represent minor respiratory aberrations, as reflected by a tendency toward carbon dioxide retention by the second twin. Route of delivery, time interval between deliveries, and nonvertex presentations were not associated with significant deviations from these observed acid-base patterns.


Joint custody and shared parenting. Research and interventions.
Joint custody is a reality for many children of divorce. This article first examines the structure and forms joint custody takes, and then explores what is known about shared parenting from research that is relevant to clinical work with families of divorce. An overview of interventions for children and families of divorce that are relevant to coparenting issues follows, along with what is known about their effectiveness. The final section presents several modes of intervention to assist families in maximizing the effectiveness of shared parenting situations for children and their parents.