Serum zinc levels and goitre epidemiology in Germany.
Zinc is an essential element involved in many basic biochemical reactions. Animal experiments and clinical data support the hypothesis that in the presence of inadequate iodine supply zinc deficiency may be a stimulus for the development of goitre. Within the framework of a study to record goitre prevalence and iodine supply of Germany we determined the serum zinc levels in 5932 clinically healthy persons. The mean value was 13.1 +/- 1.75 mumol/l. There were no differences neither in age, in sex nor in regional distribution. Furthermore there were no correlations between serum zinc level, thyroid volume and urinary iodide excretion. Only 1% of all cases had low zinc levels (< 10 mumol/l). Older people (> 40 years) with lowered serum zinc levels showed significantly larger thyroids. We conclude that there is no relevant undersupply of zinc and therefore the trace element is not involved in the goitre endemy of Germany.


Positive affect and fatigue as predictors of anti-inflammatory IL-10 concentrations among colorectal cancer patients during adjuvant chemotherapy.
(1) To examine the relationships of positive and negative affect and symptoms of depression, anxiety, and fatigue at baseline with the anti-inflammatory cytokine IL-10 concentrations in serum at three points in colorectal cancer patients; and (2) to assess the relationship between these factors and disease recurrence or mortality after a median follow-up of 24 months. In a prospective trial, 92 stage II or III colorectal cancer patients scheduled to receive standard chemotherapy were enrolled. Blood samples were collected prior to start of chemotherapy onset (T0), 3 months later (T1), and upon chemotherapy completion (T2). IL-10 concentrations were similar across the time points. Linear mixed-effects model analysis showed that controlling for confounders, higher positive affect and lower fatigue pretreatment (T0) predicted IL-10 concentrations across the time points (estimate = 0.18, SE = 0.08, 95% CI = 0.03, 0.34, p < .04 and estimate = -0.25, SE = 0.12, 95% CI = -0.50, 0.01, p < .04, respectively). Depression at T0 significantly predicted higher disease recurrence and mortality (estimate = 0.17, SE = 0.08, adjusted OR = 1.18, 95% CI = 1.02, 1.38, p = .03). We report on associations not previously assessed between positive affect and fatigue and the anti-inflammatory cytokine IL-10. Results add to previous findings suggesting that positive affect and fatigue could have a role in anti-inflammatory cytokine dysregulation.


Enzymatic hydrolysis of long-chain N-heterocyclic fatty esters.
Incubation of a 1-pyrroline ester [viz. methyl 8-(5-hexyl-1-pyrroline-2-yl)octanoate, 1] with bakers' yeast (Saccharomyces cerevisiae) gave the corresponding free fatty acid (1a, 52%). The C = N bond of the 1-pyrroline was not reduced by the yeast. Complete hydrolysis of compound 1 was successful using lipase of Candida cylindracea (CCL) or Lipolase (Rhizomucor miehei) under stirred or ultrasound condition. Fatty esters containing a pyrrolidine [viz. methyl 8-(cis/trans-5-hexyl-pyrrolidine-2-)octanoate, 2] or N-methyl pyrrolidine [viz. methyl 8-(cis-5-hexyl-N-methyl-pyrrolidine-2-)octanoate, 3] system in the alkyl chain were not hydrolyzed by either CCL or Lipolase, unless conducted in an ultrasonic bath. The hydrolytic activities of the enzymes appeared to be strongly affected by the stereochemistry of the N-heterocyclic ring system. Chemical hydrolysis of compounds 1-3 gave the corresponding fatty acid N-HCl salts.


The evaluation of evidence in the forensic investigation of fire incidents (Part I): an approach using Bayesian networks.
The forensic investigation of the origin and cause of a fire incident is a particularly demanding area of expertise. As the available evidence is often incomplete or vague, uncertainty is a key element. The present study is an attempt to approach this through the use of Bayesian networks, which have been found useful in assisting human reasoning in a variety of disciplines in which uncertainty plays a central role. The present paper describes the construction of a Bayesian network (BN) and its use for drawing inferences about propositions of interest, based upon a single, possibly non replicable item of evidence: detected residual quantities of a flammable liquid in fire debris.


Mononuclear cell infiltrate and HLA-Dr expression in intra- and extradural secondary carcinomas.
Frozen sections from 30 secondary carcinomas (22 intra- and 8 extradural) were reacted with a panel of monoclonal antibodies to macrophages, lymphocytes, NK cells and HLA-Dr invariant chain. A moderate number of macrophages was demonstrated in 91% of tumours. CD 8 and CD 4 lymphocytes were detected in smaller numbers in 68% and 28% of tumours respectively. B lymphocytes were present in only one tumour and NK cells were absent. There was no significant difference between the mononuclear cell infiltrate in the intra- and extradural tumours and in respect to the histological tumour type. HLA-Dr antigen was expressed by macrophages in most cases and by tumour cells in 5/22 (22%) (4 adeno and 1 anaplastic carcinoma; of which one adenocarcinoma was extradural and the remaining intradural). The results may represent evidence for a degree of cellular immune response to secondary carcinomas which is independent of the intra- or extradural location of the tumour.


Early cardiac changes related to radiation therapy.
To investigate the incidence and severity of possible radiation-induced cardiac changes, 21 women without heart disease were investigated serially by echocardiography and by measuring systolic time intervals before and up to 6 months after postoperative radiation therapy because of breast cancer. Radiation was associated with a decrease in fractional systolic shortening of the left ventricular (LV) minor-axis diameter, from 0.35 +/- 0.05 to 0.32 +/- 0.06 (p less than 0.005), and in the systolic blood pressure/end-systolic diameter ratio, from 4.4 +/- 1.2 to 3.9 +/- 0.9 mm Hg/mm (p less than 0.005). The mitral E point-septal separation increased, from 2.8 +/- 1.5 to 4.2 +/- 2.5 mm (p less than 0.005). The preejection period/LV ejection time ratio of systolic time intervals increased, but only the decrease within 6 months after therapy was significant (p less than 0.005). All these changes reflect slight transient depression of LV function, which became normalized within 6 months after therapy. Up to 6 months after therapy, a slight pericardial effusion was found in 33% of the patients. Hence, conventional radiation therapy appeared to cause an acute transient and usually symptomless decrease in LV function, and later, slight pericardial effusion in one-third of the patients.


Low glycemic index diet in children and young adults with refractory epilepsy: first Italian experience.
This is the report on the first Italian experience with the low glycemic index diet (LGIT) in a group of children, adolescents and young adults with refractory epileptic encephalopathies. A retrospective chart review was performed on patients initiating the LGIT in an outpatient setting from 2005 to 2010. Demographic and clinical information including seizure type, baseline seizure frequency, medications, blood chemistry, side effects, and anthropometrics were collected. Patients were educated and followed by a dietician to restrict foods with high glycemic index and to limit total daily carbohydrates to 40-60g. Change in seizure frequency was assessed at each 3-month follow-up intervals in the first year and then at each 6-month intervals. Fifteen consecutive patients (13 males and 2 females, aged between 11.3 years and 22 years), almost all affected by generalized cryptogenic or symptomatic refractory epilepsy, were enrolled in the study. After a mean follow-up period of 14.5±6.5 months (median 12.0; range 1-60 months), 6 patients (40%) had a 75-90% seizure reduction, while seizures decreased by 50% in other 2 (13.3%) and were unchanged in 7 (46.7%). The diet was discontinued in 4 patients within the first 5 months. No adverse events occurred during the diet. In conclusion, this initial experience confirms that some refractory patients may improve on the LGIT, even as first dietary option.


Clinical-pathological issues in thyroid pathology: study on the routine application of NIFTP diagnostic criteria.
In 2017, the WHO classification of tumours of the endocrine organs established the criteria for a NIFTP diagnosis. The present paper considers some aspects that are still debated or unresolved: the real incidence and clinical meaning of multifocal/multinodular lesions, the biological behaviour of micro-NIFTP, the sprinkling phenomenon and the corresponding modifications to the FNA reporting systems based on changes to the ROM. Moreover, the paper suggests possible scenarios for the clinical-pathological management of this entity. From the initial 1470 cases, a group of 68 NIFTPs was recruited in a 9 year-long period. The average age at diagnosis was 55 years. The average diameter of the lesion was 1.7 cm (0.1 cm-10 cm). In 41 cases (60.1%), the lesion was inserted in the context of a multinodular background. In 12 cases, the diagnosis was incidental and the pre- operative FNA was performed on a different target. In 10 out of 68 cases, there was a multifocal NIFTP; in 14.7% of patients, PTC-like nuclear features showed sprinkling phenomenon. The cytological revision allocated 21 cases (49%) to the SIAPEC TIR3 indeterminate class and a nuclear score 2 or 3 were identified in 25 smears. Multifocality is part of the spectrum of NIFTPs, that can arise in a multinodular background with variable sizes from microscopic lesions to very large ones. Cytopathological criteria such as an evaluation of the nuclear score may help the pathologists in promoting a NIFTP diagnosis in the preoperative setting.


Pharmacokinetic Modeling of the Glucoregulatory System.
A pharmacokinetic model is proposed to describe the glucoregulatory process. The model describes the dynamics of glucose, amino acids, and fatty acids, as well as both the hormonal actions and dynamics of insulin, glucagon, epinephrine, and glucagon-like peptide-one. The model was developed assuming that the dynamics of each species occurrs in only one compartment. Several forms of the metabolic absorption and elimination rates, along with possibilities for increasing the complexity of each compartmental model are discussed. Once properly identified and validated, the novel model has the potential to be more descriptive than other models describing glucose dynamics in the body.


Measurement error in statistical models of shape.
Active shape models (ASMs) are popular and sophisticated methods of extracting features in (especially medical) images. Here we analyse the error in placing ASM points on the boundary of the feature. By using replications, a corrected covariance matrix is presented that should reduce the effects of placement error. We show analytically and via simulations that the cumulative variability for a given number of eigenvalues retained in principal components analysis (PCA) ought to be reduced by increasing levels of point-placement error. Results for predicted errors are in excellent agreement with the set-up parameters of two simulated shapes and with anecdotal evidence from the trained experts for real data taken from the OSTEODENT project. We derive an equation for the reliability of placing the points and we find values of 0.79 and 0.85 (where 0=bad and 1=good) for the two clinical experts for the OSTEODENT data. These analyses help us to understand the sources and effects of measurement error in shape models.


Augmentation of human neutrophil and alveolar macrophage LTB4 production by N-acetylcysteine: role of hydrogen peroxide.
1. The actions of N-acetylcysteine (NAC) on hydrogen peroxide (H2O2) and leukotriene B4 (LTB4) production by human resting and stimulated peripheral blood neutrophils and alveolar macrophages were investigated. 2. At a concentration of 100 microM, NAC significantly (P < 0.01) suppressed the accumulation of H2O2 in the incubation medium of resting and opsonized zymosan (OZ; 0.5 mg ml[-1])- or N-formylmethionyl-leucyl-phenylalanine (fMLP; 1 microM)-stimulated neutrophils and of resting and OZ-stimulated macrophages. At concentrations of 10 microM and above, NAC augmented significantly the level of LTB4 in the supernatants of OZ- and fMLP-stimulated neutrophils (P < 0.01 and P < 0.05, respectively) and OZ-stimulated macrophages (P < 0.05 at 10 microM, P < 0.01 at 100 microM NAC). 3. NAC (100 microM) caused a significant (P < 0.01) reduction in the quantity of measurable H2O2 when incubated with exogenous H2O2 concentrations equivalent to those released from OZ-stimulated neutrophils and macrophages. At no concentration did NAC affect quantitites of measurable LTB4 when incubated with exogenous LTB4. 4. Superoxide dismutase (SOD), which catalyzes the conversion of superoxide anion to H2O2 had no significant effect on LTB4 production by human neutrophils. In contrast, catalase, which catalyzes the conversion of H2O2 to H2O and O2, caused a pronounced, statistically significant (P < 0.01) increase in the levels of LTB4 measured in the supernatants of OZ- and fMLP-stimulated neutrophils. 5. H2O2 (12.5 microM and 25 microM, concentrations equivalent to those measured in the supernatants of activated neutrophils and alveolar macrophages, respectively) caused a small (13%) decrease in the quantity of measurable LTB4 (P = 0.051 and P < 0.05 at 12.5 microM and 25 microM, respectively) that was inhibited by NAC (100 microM) but not by catalase (400 u ml[-1]). 6. In conclusion, the anti-oxidant drug, NAC, increases LTB4 production by human neutrophils and alveolar macrophages, probably through the elimination of cell-derived H2O2. LTB4 undergoes a H2O2-dependent oxidation that is inhibited by NAC but this is unlikely to account fully for the increased levels of LTB4, suggesting that NAC may increase LTB4 production by blocking the H2O2-dependent inhibition of a synthetic enzyme, such as 5-lipoxygenase.


Neovascular age-related macular degeneration in which exudation predominantly occurs as a subretinal fluid during anti-vascular endothelial growth factor treatment.
We investigated the characteristics of neovascular age-related macular degeneration (AMD) in which exudation predominantly occurs as a subretinal fluid (SRF) during anti-vascular endothelial growth factor (VEGF) treatment. A total of 509 treatment-naïve neovascular AMD patients treated with anti-VEGF for 24 months were retrospectively analyzed. The baseline characteristics to determine the odds of occurrence of SRF alone were evaluated using multivariate modeling. SRF was the sole manifestation of lesion activity in 209 (40.9%) eyes during follow-up. The visual outcome of eyes with only SRF occurrence during follow-up was comparable to that of eyes without exudative recurrence. In addition, the incidence of macular atrophy was significantly lower in eyes with only SRF occurrence (9.6%, 20 of 208 eyes) than in eyes without exudative recurrence (16.7%, 9 of 54 eyes, P = 0.018). Multivariate analysis revealed that better best-corrected visual acuity (BCVA) at baseline (odds ratio [OR], 0.306; P = 0.001), presence of SRF alone at baseline (OR, 5.256; P < 0.001), lower pigment epithelial detachment (PED) height (less than 100 µm; OR, 4.113; P = 0.025), and aneurysmal type 1 macular neovascularization (MNV) (OR, 2.594; P = 0.002) were associated with an increased likelihood of SRF occurrence during follow-up. In conclusion, the eyes with only SRF occurrence during anti-VEGF treatment showed more favorable visual outcomes and a lower incidence of macular atrophy. The baseline characteristics, including better baseline BCVA, presence of SRF alone at baseline, lower PED height, and MNV subtype, might influence the predominant development of SRF during anti-VEGF treatment.


Lipid domain in cancer cell plasma membrane shown by 1H NMR to be similar to a lipoprotein.
Human blood lipoproteins have been characterised by 1H NMR methods and chemical analysis, and comparisons made with the properties of the triglyceride-rich plasma membrane domain found in cancer cells. By means of selective and non-selective T1 experiments, the lipids in HDL and LDL are shown to be in diffusive exchange. In contrast, the lipids of chylomicra and VLDL do not exhibit lipid diffusion, and therefore resemble the neutral lipids of cancer cell plasma membranes. 2D scalar correlated NMR (COSY) spectra of cancer cells or solid tumours are similar to those obtained from VLDL and LDL. The long T2 relaxation value observed for neutral lipid methylenes in metastatic cancer cells (greater than 300 ms) was not observed for any of the 4 lipoproteins studied. None of the lipoprotein classes gave a T2 longer than 250 ms.


Exome sequencing reveled a compound heterozygous mutations in RTTN gene causing developmental delay and primary microcephaly.
RTTN (Rotatin) (OMIM 614833) is a large centrosomal protein coding gene. RTTN mutations are responsible for syndromic forms of malformation of brain development, leading to polymicrogyria, microcephaly, primordial dwarfism, seizure along with many other malformations. In this study we have identified a compound heterozygous mutation in RTTN gene having NM_173630 c.5225A > G p.His1742Arg in exon 39 and NM_173630 c.6038G > T p.Cys2013Phe in exon 45 of a consanguineous Saudi family leading to brain malformation, seizure, developmental delay, dysmorphic feature and microcephaly. Whole exome sequencing (WES) techniques was used to identify the causative mutation in the affected members of the family. WES data analysis was done and obtained data were further confirmed by using Sanger sequencing analysis. Moreover, the mutation was ruled out in 100 healthy control from normal population. To the best of our knowledge the novel compound heterozygous mutation observed in this study is the first report from Saudi Arabia. The identified compound heterozygous mutation will further explain the role of RTTN gene in development of microcephaly and neurodevelopmental disorders.


Disseminated cutaneous sporotrichosis. Three illustrative cases.
Three cases of disseminated cutaneous sporotrichosis are reported, including a rare presentation of the disease as vesicular dermatitis. The diagnosis was confirmed by culture of a skin biopsy specimen, and all patients responded well to amphotericin B therapy. The sporotrichosis slide latex agglutination titer was used to determine the end point of therapy.


Evaluation of different protocols for culturing mesenchymal stem cells derived from murine bone marrow.
Culturing bone marrow mesenchymal stem cells (BM-MSCs) is a key point in different fields of research, including tissue engineering and regenerative medicine and studies of the bone marrow microenvironment. However, isolating and expanding murine BM-MSCs in vitro has challenged researchers due to the low purity and yield of obtained cells. In this study, we aimed to evaluate five different protocols to culture murine BM-MSCs in vitro. All protocols were based on the adhesion capacity of BM-MSCs to the tissue culture plastic surface and varied in the types of plate, culture media, serum, additional supplementation and initial cell density. Flow cytometry analysis was used to investigate lineage purity after expansion. The expression of CD45 and CD11b was detected in the cultures generated according to all protocols, indicating low purity with the presence of hematopoietic cells and macrophages. The cellular growth rate and morphology varied between the cultures performed according to each protocol. Cells cultured according to protocol 5 (8 × 107cells/plate, Roswell Park Memorial Institute (RPMI) culture medium during first passage and then Iscove's Modified Delbecco's Medium (IMDM) culture medium, both supplemented with 9% fetal bovine serum, 9% horse serum, 12µM L-glutamine) presented the best performance, with a satisfactory growth rate and spindle-shape morphology. Our results point out that the purity and satisfactory growth rate of murine BM-MSC cultures are not easily achieved and additional approaches must be tested for a proper cell expansion.


Displacement of the optic nerve head by acute changes in intraocular pressure in monkey eyes.
The authors used the Humphrey Retinal Analyzer to study the effect of acute changes in intraocular pressure (IOP) on the topography of the optic nerve head in normotensive and hypertensive eyes of cynomolgus monkeys. Chronically elevated IOP was produced monocularly in monkeys by argon laser angle treatment. In 8 hypertensive eyes, the mean IOP of 51 mmHg was lowered acutely to 15 mmHg with a needle placed in the anterior chamber. In 12 normotensive eyes, the mean IOP of 16 mmHg was similarly increased to 45 mmHg. Simultaneous stereophotographs were taken both before and within 15 minutes after the IOP change. The surface of normotensive optic nerve heads moved posteriorly a mean of 17.8 microns after IOP elevation (P less than 0.02). The surface of hypertensive optic nerve heads moved anteriorly 47.4 microns after IOP lowering (P = 0.1); this change was significantly less in nerve heads with larger and deeper cups (P less than 0.02). Significant changes were not detected in disc diameter or vein diameter with acute changes in IOP. Measurable shifts in the position of the optic nerve head surface when IOP is altered may provide a future prognostic test for glaucomatous optic neuropathy.


Functional gastrointestinal disorders in children: agreement between Rome III and Rome IV diagnoses.
To evaluate the agreement between the Rome III and Rome IV criteria in diagnosing pediatric functional gastrointestinal disorders (FGIDs), we conducted a prospective cohort study in a public school in Cali, Colombia. Children and adolescents between 11 and 18 years of age were given the Spanish version of the Questionnaire on Pediatric Functional Gastrointestinal Disorders Rome III version on day 0 and Rome IV version on day 2 (48 h later). The study protocol was completed by 135 children. Thirty-nine (28.9%) children were excluded because of not following the instructions of the questionnaire. The final analysis included data of 96 children (mean 15.2 years old, SD ± 1.7, 54% girls). Less children fulfilled the criteria for an FGID according to Rome IV compared to Rome III (40.6% vs 29.2%, p=0.063) resulting in a minimal agreement between the two criteria in diagnosing an FGID (kappa 0.34, agreement of 70%). The prevalence of functional constipation according to Rome IV was significantly lower compared to Rome III (13.5% vs 31.3%, p<0.001), whereas functional dyspepsia had a higher prevalence according to Rome IV than Rome III (11.5% vs 0%).Conclusion: We found an overall minimal agreement in diagnosing FGIDs according to Rome III and Rome IV criteria. This may be partly explained by the differences in diagnostic criteria. However, limitations with the use of questionnaires to measure prevalence have to be taken into account. What is Known: • The Rome IV criteria replaced the previous Rome III criteria providing updated criteria to diagnose functional gastrointestinal disorders (FGIDs). • Differences found between Rome IV and historic Rome III FGID prevalence may have been affected by changes in prevalence over time or differences in sample characteristics. What is New: • We found a minimal agreement between Rome III and Rome IV FGID diagnosis, especially in the diagnoses of functional constipation, irritable bowel syndrome, and functional dyspepsia. • The minimal agreement may be partly explained by changes in diagnostic criteria, but limitations with the use of questionnaires to measure prevalence have to be taken into account.


Initial experience of endoscopic ultrasound-guided antegrade covered stent placement with long duodenal extension for malignant distal biliary obstruction (with video).
This study aimed to evaluate the feasibility of endoscopic ultrasound (EUS)-guided antegrade covered stent placement with long duodenal extension (EASL) for malignant distal biliary obstruction (MDBO) with duodenal obstruction (DO) or surgically altered anatomy (SAA) after failed endoscopic retrograde cholangiopancreatography (ERCP). Outcomes were technical and clinical success, reintervention rate, adverse events, stent patency, and overall survival. Inverse probability of treatment weighting (IPTW) and competing-risk analysis were performed to compare with conventional EUS-BD. Twenty-five patients (DO, n = 18; SAA, n = 7) were included. The technical and clinical success rates were 96% and 84%, respectively. Reintervention occurred in two patients (8.3%). Adverse events occurred in six patients (24%; two cholangitis, 16%; four mild postprocedural pancreatitis [24% (n = 4/17) in patients with non-pancreatic cancers]). The median patency was 9.4 months, and the overall survival was 2.73 months. After IPTW adjustment, the median patency in the EASL (n = 25) and conventional EUS-BD (n = 29) were 10.1 and 6.5 months, respectively (P = .018). EASL has acceptable clinical outcomes with a low reintervention rate but higher rate of postprocedural pancreatitis in patients with non-pancreatic cancers. Randomized trials comparing EASL and conventional EUS-BD for MDBO with pancreatic cancers and DO/SAA after failed ERCP are needed to validate our findings.


A silicon-glass microwell platform for high-resolution imaging and high-content screening with single cell resolution.
We present a novel microwell array platform suited for various cell-imaging assays where single cell resolution is important. The platform consists of an exchangeable silicon-glass microchip for cell biological applications and a custom made holder that fits in conventional microscopes. The microchips presented here contain arrays of miniature wells, where the well sizes and layout have been designed for different applications, including single cell imaging, studies of cell-cell interactions or ultrasonic manipulation of cells. The device has been designed to be easy to use, to allow long-term assays (spanning several days) with read-outs based on high-resolution imaging or high-content screening. This study is focused on screening applications and an automatic cell counting protocol is described and evaluated. Finally, we have tested the device and automatic counting by studying the selective survival and clonal expansion of 721.221 B cells transfected to express HLA Cw6-GFP compared to untransfected 721.221 B cells when grown under antibiotic selection for 3 days. The device and automated analysis protocol make up the foundation for development of several novel cellular imaging assays.


Umbilical cord androgens and estrogens in relation to verbal and nonverbal abilities at age 10 in the general population.
Sex differences in verbal and nonverbal abilities are a contentious area of research. Prenatal steroids have been shown to have masculinizing effects on the brain that may affect the development of nonverbal and verbal abilities in later life. The current study examined a wide range of biologically active sex steroids (both androgens and estrogens) in umbilical cord blood at birth in a large pregnancy cohort in relation to performance on nonverbal (Raven's Coloured Progressive Matrices) and verbal (Clinical Evaluation of Language Fundamentals-3 and the Peabody Picture Vocabulary Test-III) measures at age 10 years. Overall, Androgen and Estrogen composites in cord blood were not found to be predictive of performance on verbal and nonverbal measures at age 10. These data suggest that late gestation sex steroids do not exert a major effect on nonverbal and verbal abilities in middle childhood.


Significantly improved precision of cell migration analysis in time-lapse video microscopy through use of a fully automated tracking system.
Cell motility is a critical parameter in many physiological as well as pathophysiological processes. In time-lapse video microscopy, manual cell tracking remains the most common method of analyzing migratory behavior of cell populations. In addition to being labor-intensive, this method is susceptible to user-dependent errors regarding the selection of "representative" subsets of cells and manual determination of precise cell positions. We have quantitatively analyzed these error sources, demonstrating that manual cell tracking of pancreatic cancer cells lead to mis-calculation of migration rates of up to 410%. In order to provide for objective measurements of cell migration rates, we have employed multi-target tracking technologies commonly used in radar applications to develop fully automated cell identification and tracking system suitable for high throughput screening of video sequences of unstained living cells. We demonstrate that our automatic multi target tracking system identifies cell objects, follows individual cells and computes migration rates with high precision, clearly outperforming manual procedures.


Capnocytophaga in the amniotic fluid of a woman in preterm labor with intact membranes.
Chorioamnionitis is thought to be uncommon with intact fetal membranes. A case is reported of a woman in preterm labor with intact membranes from whose amniotic fluid, placenta, and infant's gastric aspirate an unusual organism, Capnocytophaga, was identified. The findings are discussed.


[Hospital inhouse contact tracing-significant impact to patient care during the coronavirus pandemic].
As part of the SARS-CoV‑2 pandemic, the district of Heinsberg developed into an infectiological epicentre for Germany in February 2020. Our hospital, which is located in the immediate vicinity, reacted very quickly in addition to adapting patient care by implementing an organizational structure for recording SARS-CoV-2-positive employees, patients and their contact persons. The infections recorded in contact tracing were analysed and, based on an exemplary outbreak, infection chains and follow-up processes were evaluated. Comprehensive data on contact types, oropharyngeal swab results for SARS-CoV‑2 and quarantine days were documented and retrospectively evaluated using a self-developed database. Of the 568 employees recorded by in-house contact tracing, 32 employees (1.2%, n = 2567) were detected as SARS-CoV‑2 positive. Of those, 50% (n = 16) tested positive due to contact tracing, 15.6% (n = 5) were recorded by routine smears and 34.4% (n = 11) were returning travellers. The variable PCR results of the control smears from these positive employees were noticeable. In 18.8% (n = 6) of the initially negative control smears, positive PCR results were found in the following control smear. The inhouse contact tracing team was able to detect infection clusters on non-COVID-19 wards at an early stage and, together with clinical hygiene and the public health department, initiated comprehensive measures to limit the spread of the virus. Infection chains could thus be interrupted. The work of the clinic's own contact tracing unit has proven to be an essential part of clinical pandemic management not least against the background of new waves of infection and is indispensable for the detection of local infection clusters.


Carbohydrate-binding motif in chitinase 3-like 1 (CHI3L1/YKL-40) specifically activates Akt signaling pathway in colonic epithelial cells.
Host-microbial interactions play a key role during the development of colitis. We have previously shown that chinase 3-like 1 (CHI3L1) is an inducible molecule overexpressed in colonic epithelial cells (CECs) under inflammatory conditions. In this study, we found that chitin-binding motif (CBM) of CHI3L1 is specifically associated with the CHI3L1-mediated activation of the Akt-signaling in CEC by transfecting the CBM-mutant CHI3L1 vectors in SW480 CECs. Downstream, CHI3L1 enhanced the secretion of IL-8 and TNFα in a dose-dependent manner. We previously show that 325 through 339 amino-acids in CBM are crucial for the biological function of CHI3L1. Here we demonstrated that 325th-339th residues of CBM in CHI3L1 is a critical region for the activation of Akt, IL-8 production, and for a specific cellular localization of CHI3L1. In conclusion, CBM region of CHI3L1 is critical in activating Akt signaling in CECs, and the activation may be associated with the development of chronic colitis.


Spontaneous thermal motion of the GABA(A) receptor M2 channel-lining segments.
The gamma-aminobutyric acid type A (GABA(A)) receptor channel opening involves translational and rotational motions of the five channel-lining, M2 transmembrane segments. The M2 segment's extracellular half is loosely packed and undergoes significant thermal motion. To characterize the extent of the M2 segment's motion, we used disulfide trapping experiments between pairs of engineered cysteines. In alpha1beta1 gamma2S receptors the single gamma subunit is flanked by an alpha and beta subunit. The gamma2 M2-14' position is located in the alpha-gamma subunit interface. Gamma2 13' faces the channel lumen. We expressed either the gamma2 14' or the gamma2 13' cysteine substitution mutants with alpha1 cysteine substitution mutants between 12' and 16' and wild-type beta1. Disulfide bonds formed spontaneously between gamma2 14'C and both alpha1 15'C and alpha1 16'C and also between gamma2 13'C and alpha1 13'C. Oxidation by copper phenanthroline induced disulfide bond formation between gamma2 14'C and alpha1 13'C. Disulfide bond formation rates with gamma2 14'C were similar in the presence and absence of GABA, although the rate with alpha1 13'C was slower than with the other two positions. In a homology model based on the acetylcholine receptor structure, alphaM2 would need to rotate in opposite directions by approximately 80 degrees to bring alpha1 13' and alpha1 15' into close proximity with gamma2 14'. Alternatively, translational motion of alphaM2 would reduce the extent of rotational motion necessary to bring these two alpha subunit residues into close proximity with the gamma2 14' position. These experiments demonstrate that in the closed state the M2 segments undergo continuous spontaneous motion in the region near the extracellular end of the channel gate. Opening the gate may involve similar but concerted motions of the M2 segments.


Dissemination of metallo-β-lactamase in Pseudomonas aeruginosa isolates in Egypt: mutation in blaVIM-4.
This study was designed to investigate the prevalence of metallo-β-lactamase (MBL) in Pseudomonas aeruginosa isolates collected from Suez Canal University Hospital in Ismailia, Egypt. Antibiotic susceptibility testing and phenotypic and genotypic screening for MBLs were performed on 147 isolates of P. aeruginosa. MICs were determined by agar dilution method for carbapenem that was ≥2 μg/mL for meropenem. MBL genes were detected by multiplex and monoplex PCR for P. aeruginosa-harbored plasmids. Mutation profile of sequenced MBL genes was screened using online software Clustal Omega. Out of 147 P. aeruginosa, 39 (26.5%) were carbapenem-resistant isolates and 25 (64%) were confirmed to be positive for MBLs. The susceptibility rate of P. aeruginosa toward polymyxin B and norfloxacin was 99% and 88%, respectively. Identification of collected isolates by API analysis and constructed phylogenetic tree of 16S rRNA showed that the isolates were related to P. aeruginosa species. The frequency of blaGIM-1, blaSIM-1, and blaSPM-1 was 52%, 48%, and 24%, respectively. BlaVIM and blaIMP-like genes were 20% and 4% and the sequences confirm the isolate to be blaVIM-1, blaVIM-2, blaVIM-4, and blaIMP-1. Three mutations were identified in blaVIM-4 gene. Our study emphasizes the high occurrence of multidrug-resistant P. aeruginosa-producing MBL enzymes.


Correction to: EVERYTHING HAS CHANGED FOR THE WORSE: EXPERIENCES OF ELDERS DISPLACED BY ARMED CONFLICT IN ETHIOPIA.
[This corrects the article DOI: 10.1093/geroni/igac059.2131.].


Vitrectomy for epiretinal membrane with Candida chorioretinitis.
Four eyes underwent vitrectomy and epiretinal membrane dissection for tractional retinal detachment (RD) or macular pucker associated with Candida chorioretinitis. The epiretinal membranes were vascularized and, in three of four eyes, were associated with full-thickness retinal scars, presumably at the site of previous active Candida chorioretinitis. The epiretinal membranes were successfully removed in all four cases. Visual results depended on the degree of macular pathology and the presence and location of a full-thickness retinal scar.


Facial morphology and malocclusions.
The aim of this study was to determine whether sagittal and vertical aberrations in intermaxillary incisal tooth and jaw relationships were reflected in facial morphology. One hundred seventy-two children (79 girls and 93 boys), 12 to 14 years of age, were selected from the Orthodontic Department, University of Giessen. Pretreatment profile and frontal facial photographs as well as profile roentgenograms in centric occlusion for each child were made. The method consisted of (1) a personal appraisal of the existing incisal tooth and jaw relationship by visual inspection of facial photographs, (2) a validity assessment of these personal appraisals, and (3) a comparison of linear and angular measurements made from facial photographs and from lateral headfilms. The result of this investigation revealed that a large overjet and a Class II, Division 2 incisal tooth relationship were most often reflected in the face. A Class III incisal tooth relationship and an open bite, however, were difficult to detect. Sagittal maxillary and mandibular positions could only occasionally be determined on the basis of facial appearance, while a Class II jaw base relationship (large ANB angle) could easily be seen. The vertical jaw relationship (mandibular plane angle and anterior facial height index) could be determined with relatively high precision. When the measurements from the facial photographs were compared with those from the lateral headfilms, moderate to high correlations were found between skeletal and soft tissue readings: ANB angle (r = +0.63), mandibular plane angle (r = +0.93), and anterior facial height index (r = +0.86). It was concluded that sagittal and vertical dental and skeletal intermaxillary malrelationships were only partly reflected in the face.


[Magnetic Drug Targeting--a new approach in locoregional tumor therapy with chemotherapeutic agents. Experimental animal studies].
Advanced squamous cell carcinomas of the head and neck region were often treated with combined radio-chemotherapy. Radiotherapy allows a focused treatment of the tumor, and healthy tissue can be protected from radiation. Chemotherapy, however, is mostly given systemically and the unwanted negative side effects also develop in many other organs. Locoregional application of chemotherapeutic agents with Magnetic Drug Targeting on an animal experimental study. Magnetic Drug Targeting is a new approach to the locoregional treatment of tumors. Ferrofluids (colloidal dispersion of magnetic nanoparticles) were reversibly bound to chemotherapeutic agents and injected intra-arterially, while focused with an external magnetic field to a certain body compartment (i.e. the tumor). With only 20% or 50% percent of the regular systemic chemotherapeutic dose, we achieved an up to 26 times higher concentration in the tumor region with this application compared to the usual systemic administration. Magnetic Drug Targeting offers an unique opportunity to treat tumors locoregionally with chemotherapeutic agents.


Percent Insular Ribbon Infarction for Predicting Infarct Growth Rate and 90-Day Outcomes in Large-Vessel Occlusive Stroke: Secondary Analysis of Prospective Clinical Trial Data.
BACKGROUND. Insight into the natural history of infarct growth could help identify patients with slowly progressing stroke who may benefit from delayed endovascular thrombectomy (EVT). OBJECTIVE. The purpose of this article is to evaluate associations of percent insular ribbon infarction (PIRI) with infarct growth rate (IGR) and 90-day outcomes in patients with large-vessel occlusive stroke. METHODS. This retrospective study was a secondary analysis of a prior clinical trial that enrolled patients with acute stroke not treated with reperfusion therapies from January 2007 to June 2009. The present analysis evaluated 31 trial patients (median age, 71 years; 12 women, 19 men) with anterior-circulation large-vessel occlusion who underwent serial MRI examinations. Two neuroradiologists independently scored PIRI on presentation MRI examinations on the basis of the ratio of the length of the portion of the insula showing restricted diffusion to the insula's total length using a previously described 0-4 scale; scores were categorized (mild [0-1], moderate [2], or severe [3-4]), and discrepancies were resolved by consensus. The 90-day modified Rankin Scale (mRS) was obtained. As part of earlier clinical trial analyses, collateral pattern on CTA was classified as symmetric, malignant, or other, and infarct volumes were measured on DWI during the initial 48 hours after presentation and on FLAIR at 90 days. RESULTS. Interrater agreement for PIRI category was strong (κ = 0.89). PIRI was mild in 10, moderate in four, and severe in 17 patients. For mild, moderate, and severe PIRI, median IGR from onset to presentation was 1.6 cm3/h, 8.5 cm3/h, and 17.5 cm3/h (p < .001); median IGR from presentation to 48 hours was 0.3 cm3/h, 0.2 cm3/h, and 1.2 cm3/h (p = .005); median 90-day infarct volume was 9.4 cm3, 39.8 cm3, and 108.6 cm3 (p = .01); and 90-day mRS of 2 or less occurred in 78%, 67%, and 6% of patients (p = .001). In multivariable models controlling for age, internal carotid artery occlusion, and collateral pattern, PIRI category independently predicted onset-to-presentation IGR (β = 1.5), presentation-to-48-hour IGR (β = 1.3), and 90-day mRS of 2 or less (OR = 0.2). For predicting 90-day mRS of 2 or less, mild-to-moderate PIRI had sensitivity of 90.0% and specificity of 84.2%; symmetric collateral pattern had sensitivity of 70.0% and specificity of 73.7%. CONCLUSION. PIRI was independently associated with IGR and 90-day outcome. CLINICAL IMPACT. PIRI may help identify patients who could benefit from late-window EVT when requiring transfer to EVT-capable centers.


Perforating ocular injury caused by acupuncture: A case report.
Acpuncture as a branch of traditional Chinese medicine is popular in China. It can regulate the running of meridian qi to stimulate the ocular nerve activity and increase blood supply. Periocular acupuncture treatment is very frequent, but it can lead to safety hazards that cannot be ignored. For instance, ocular trauma may develop if done improperly, resulting in impaired vision and even blindness. We report a rare case of perforating ocular injury caused by acupuncture.


Self-Peptidome Variation Shapes Individual Immune Responses.
The relationship between human genetic variation and disease has not been fully elucidated. According to the present view on infectious diseases pathogen resistance is linked to human leukocyte antigen (HLA) class I/II variants and their individual capacity to present pathogen-derived peptides. Yet, T cell education in the thymus occurs through negative and positive selection, and both processes are controlled by a combination of HLA class I/II variants and peptides from the self. Therefore, the capacity of given HLA class I/II variants to bind pathogen-derived peptides is only one part of the selective process to generate effective immune responses. We thus propose that peptidome variation contributes to shaping T cell receptor (TCR) repertoires and hence individual immune responses, and that this variation represents inherent modulator epitopes.


Characteristics of the receptors in the isolated capsule of the hip in the cat.
The properties of the afferent fibres from the capsule of the hip joint have been studied in the cat in situ, in relations to joint rotation, and in an isolated capsule preparation which was opened and stretched directly with an actuator. In situ two types of afferent fibres were found, those having a full range of sensitivity and others having only a limited range in response to the joint rotation. When studied in isolated tissue the afferent fibres of the capsule were uniform in threshold and sensitivities, and no full range receptors were found. We conclude that the full range receptors which enter the articular nerve of the hip are spindle afferents and not capsule receptors. On the basis of these and previous results in animals and man the role of joint receptors in kinaesthesia and position sense is discussed.


Efficacy and safety of autologous adipose-derived stromal vascular fraction enriched with platelet-rich plasma in flap repair of transsphincteric cryptoglandular fistulas.
Transanal advancement flap repair of transsphincteric fistulas is a sphincter-preserving procedure, which frequently fails, probably due to ongoing inflammation in the remaining fistula tract. Adipose-derived stromal vascular fraction (SVF) has immunomodulatory properties promoting wound healing and suppressing inflammation. Platelet-rich plasma (PRP) reinforces this biological effect. The aim of this study was to evaluate the efficacy and safety of autologous adipose-derived SVF enriched with PRP in flap repair of transsphincteric cryptoglandular fistulas. A prospective cohort study was conducted including consecutive patients with transsphincteric cryptoglandular fistula in a tertiary referral center. During flap repair, SVF was obtained by lipoharvesting and mechanical fractionation of adipose tissue and combined with PRP was injected around the internal opening and into the fistulous wall. Endpoints were fistula healing at clinical examination and fistula closure on postoperative magnetic resonance imaging (MRI). Adverse events were documented. Forty-five patients with transsphincteric cryptoglandular fistula were included (29 males, median age 44 years [range 36-53 years]). In the total study population, primary fistula healing was observed in 38 patients (84%). Among the 42 patients with intestinal continuity at time of surgery, primary fistula healing was observed in 35 patients (84%). In one patient, the fistula recurred, resulting in a long-term healing rate of 82%. MRI, performed in 37 patients, revealed complete closure of the fistula tract in 33 (89.2%). In the other patients, the tract was almost completely obliterated by scar tissue. During follow-up, none of these patients showed clinical signs of recurrence. The postoperative course was uneventful, except for three cases; venous thromboembolism in one patient and bleeding under the flap, necessitating intervention in two patients. Addition of autologous SVF enriched with PRP during flap repair is feasible, safe and might improve outcomes in patients with a transsphincteric cryptoglandular fistula. Dutch Trial Register, Trial Number: NL8416, https://www.trialregister.nl/.


Targeting Viral Methyltransferases: An Approach to Antiviral Treatment for ssRNA Viruses.
Methyltransferase enzymes have been associated with different processes within cells and viruses. Specifically, within viruses, methyltransferases are used to form the 5'cap-0 structure for optimal evasion of the host innate immune system. In this paper, we seek to discuss the various methyltransferases that exist within single-stranded RNA (ssRNA) viruses along with their respective inhibitors. Additionally, the importance of motifs such as the KDKE tetrad and glycine-rich motif in the catalytic activity of methyltransferases is discussed.


Arachnoid cyst within the fourth ventricle--a case report.
We describe a case of congenital intra-fourth ventricular arachnoid cyst and review the literature. A one-and-half-year-old boy had presented with delayed milestones, intermittent raised intracranial pressure and ataxia. Excision of the arachnoid cyst has cured him.


Radiographic features of hand and wrist arthroplasties.
The normal and abnormal radiographic appearances of various arthroplasties of the hand and wrist have been presented. It is hoped that increased familiarity with these procedures and their radiographic features will lead to more meaningful radiographic interpretations by consulting radiologists.


Glossopharyngeal neuralgia treated with an endoscopic assisted midline suboccipital subtonsillar approach: technical note.
Glossopharyngeal nerve neuralgia (GPN) is an exceptionally rare skull-based disorder. It is treated similar to other neurovascular conflicts. The approach, however, is debatable, as the caudal cranial nerves and especially the exit zones at the brainstem are barely visible via the retrosigmoid approach. We demonstrate a case of a typical GPN with microsurgical neurovascular decompression according to standard procedures. Continuous electrophysiological monitoring, including motor-evoked potentials of the glossopharyngeal nerve in addition to somatosensory evoked potential (SSEP) monitoring, was set up. In prone position a midline suboccipital osteoplastic craniotomy was performed. Intradurally, an endoscopic-assisted subtonsillar exposure of the lateral recess and the nerve root exit zone of the glossopharyngeal nerve has been chosen. A large compressing branch of the posterior inferior cerebellar artery (PICA) was identified as the cause of the neurovascular conflict and transposed. A midline subtonsillar approach with endoscopic assistance to the lateral brainstem is a straightforward time- and morbidity-sparing procedure.


Development of the nursing practice scale for end-of-life family conferences in critical care.
This study aimed to develop the Nursing Practice Scale for End-of-life Family conferences in critical care and to clarify the current status of nursing practice regarding family conferences. We conducted a cross-sectional, self-administered questionnaire survey with 955 critical care unit nurses in 97 hospitals. Content validity, factor validity and criterion-related validity, known-group validity, internal consistency and test-retest reliability were evaluated. Data were then analysed statistically. Adult intensive care units or high dependency units in Japan. Three factors with 39 items were extracted through item analysis and confirmatory factor analysis as hypothesised (Factor 1: Preparation, Factor 2: Discussion and Factor 3: Follow-up), and the mean score per item for each factor was 3.57, 3.73 and 3.75, respectively. Nurses who had any certification or had worked in critical care unit for >5 years had a significantly greater score than the others. The Cronbach's α were 0.86-0.96 and the intraclass correlation coefficients were 0.79-0.87. The Nursing Practice Scale for End-of-life Family conferences in critical care is a valid and reliable scale. This study could effectively facilitate communication among patients, their families and healthcare providers.


Asymmetric Synthesis of Oxa-Bridged Oxazocines through a Catalytic RhII /ZnII Relay [4+3] Cycloaddition Reaction.
Oxa-bridged oxazocines bearing three chiral carbon centers were synthesized efficiently through a bimetallic catalytic asymmetric tandem reaction of β,γ-unsaturated α-ketoesters with diazoimides. The process contained a rhodium-promoted in situ generation of isomünchnone from diazoimide decomposition, and a [4+3]-cycloaddition of β,γ-unsaturated α-ketoester catalyzed by a chiral N,N'-dioxide-ZnII complex. Ligand-accelerated catalysis was found, and a possible transition-state model was proposed to explain the origin of stereoselectivity.


Evolutionary genomics of recent clinical Bordetella pertussis isolates from Iran: wide circulation of multiple ptxP3 lineages and report of the first ptxP3 filamentous hemagglutinin-negative B. pertussis.
Here we investigated nationwide clinical Bordetella pertussis isolated during 2005-2017 from different provinces of Iran, a country with more than 50 years whole cell vaccine immunisation history. Our results revealed the ongoing increase in the population of ptxP3/fim3-2 B. pertussis isolates in different provinces which were differentiated into nine clades. The largest clade (clade 8) which was previously found to be prevalent in Tehran was also prevalent across the country and clade 5 with ptxP3/prn9 genotype has also increased in frequency (14% of all ptxP3 isolates) in recent years. Furthermore, we detected the first ptxP3 B. pertussis isolates that does not express filamentous hemagglutinin (FhaB) as one of the major antigens of the pathogen and a key component of the acellular pertussis vaccine.


Cytochrome P-450 3A: interactions with dermatologic therapies.
Recent case reports and studies suggest that interactions involving the cytochrome P-450 mixed function oxidase system are important causes of medication toxicity and decreased efficacy during combination drug therapy. The cytochrome P-450 3A3/4 isoenzyme is involved in many significant drug interactions. New and familiar drugs continue to be implicated as having potentially serious interactions with this group of enzymes. An understanding of the basic principles of these interactions may have a major impact on patient outcome.


The health needs of Aboriginal and Torres Strait Islander children in out-of-home care.
Children in statutory out-of-home care (OOHC) are known to have poorer physical, developmental and mental health outcomes compared with their peers. The number of children in OOHC is increasing in Australia, with Aboriginal and Torres Strait Islander children disproportionately represented. To document the health needs of Aboriginal and Torres Strait Islander children and adolescents in OOHC attending the paediatric service at the Victorian Aboriginal Health Service (VAHS) between February 2014 and February 2016. The electronic medical records of all patients in OOHC seen by a paediatrician at VAHS between February 2014 and February 2016 were audited. Demographic data, including age and type of OOHC placement, were recorded, as were their health needs. Of the 200 children seen by a paediatrician over this 2-year period, 103 were identified as being in OOHC for all or some of this same period: 43 in foster care, 8 in residential care and 52 in kinship care. Of the 103 children, 35 were <6 years of age, 51 between 6 and 13 years and 17 between 13 and 18 years; 66% were identified as having mental health problems, 37% hearing problems, 34% vision problems, 40% dental caries and 46% developmental delay including 36% with speech delay. Aboriginal and Torres Strait Islander children in OOHC seen by the paediatric service at VAHS have high rates of physical, developmental and psychological health needs. These data have significant potential to inform government policy and investment in effective models of care to better meet the needs of Aboriginal and Torres Strait Islander children in OOHC.


Synthesis of planar chiral ferrocenes via enantioselective remote C-H activation.
Planar chiral ferrocenes are widely studied structures in asymmetric catalysis, materials science and medicinal chemistry. Although synthetic methods for 1,2-disubstituted planar chiral ferrocenes are well known, methods for the direct construction of 1,3-disubstituted planar chiral ferrocenes remain elusive. Here we report a modular platform for the construction of planar chirality in 1,3-disubstituted ferrocenes/ruthenocenes via an enantioselective relay remote C-H activation strategy. This method demonstrates a mechanism for remote enantiocontrol via enantiodetermining initial C‒H activation at the C2 position, enabled by a chiral mono-N-protected natural amino-acid ligand, and subsequent relay to the remote C3 position by a bridgehead-substituted norbornene mediator. A wide variety of 1,3-disubstituted planar chiral metallocenes are prepared with high enantioselectivity (96‒99% e.e.). The reaction shows good functional-group tolerance and high step-economy, and aryl iodides/bromides are compatible as coupling partners. The resulting metallocenes can be readily derivatized to yield planar chiral ligands and catalysts for asymmetric catalysis as well as building blocks for other applications.


Bilateral renal malakoplakia.
A case of malakoplakia is presented which was confined to the renal parenchyma of both kidneys in a thirty-five-year-old woman in whom fatal renal failure developed in three weeks. Bilateral renal malakoplakia is rare, and unlike malakoplakia of the bladder, it behaves as a progressive, destructive, and fatal disease.


Pneumonia surveillance and its attendant clinical risk stratification for COVID-19 in low-risk patients.
The objective of the study is to determine the prevalence of COVID-19 in the context of a secondary pneumonia surveillance program targeted at low-risk patients and to identify clinical characteristics associated with COVID-19. This study design is a retrospective cohort study. This study is conducted in Tan Tock Seng Hospital, a University affiliated 1600-bed public hospital in Singapore. Patients with pneumonia admitted under our Enhanced Pneumonia Surveillance (EPS) program from 7 February 2020 to 20 March 2020 were included. Relevant clinical variables were collated. Of 1295 patients admitted under our EPS program, 47 (3.6%) patients tested positive for COVID-19. The prevalence of a radiologist-reported normal chest X-ray (CXR) in the COVID-19-positive group was 62.8% compared with 6.2% in the COVID-19-negative group. In patients with a normal CXR, a low normal white blood cell (WBC) count and minimal C-reactive protein (CRP) elevation were associated with COVID-19. The pick-up rate of COVID-19 in low-risk patients with pneumonia is 3.6%. However, at least 7.9% of patients who were isolated had a normal CXR. For patients with pneumonia-like illness at presentation but a normal CXR, higher WBC and CRP values may guide early deisolation. Ultimately, this informs resource allocation for both COVID-19 and non-COVID-19 clinical services.


Comprehensive analysis of microRNA expression in lumbar facet joint capsules and synovium of patients with osteoarthritis: Comparison between early-stage and late-stage osteoarthritis samples from a single individual.
MicroRNA is attracting attention as a therapeutic target for osteoarthritis. We focused on joint capsules and synovium in lumbar facet joint osteoarthritis. The purpose of this study was to identify microRNAs that are upregulated in lumbar facet joint capsules and synovium with osteoarthritis. We included patients who underwent spinal fusion for degenerative lumbar spine diseases. We selected patients who had both early-stage and late-stage facet joint osteoarthritis in a single individual. We extracted joint capsule and synovium samples from these patients and isolated microRNAs. During the screening phase, we compared early-stage and late-stage osteoarthritis samples from the same individual. We identified microRNAs with >2-fold change in expression in 75% or more of patients with late-stage osteoarthritis using next generation sequencing. During the technical validation phase, the same samples were used for real-time polymerase chain reaction. We identified microRNAs with >2-fold change in expression in 62.5% or more of patients with late-stage osteoarthritis. Of 40 patients who underwent spinal fusion, we selected eight patients with both early-stage and late-stage facet joint osteoarthritis. During the screening phase, we identified eight upregulated microRNAs out of 2274 microRNAs in late-stage OA. In late-stage OA, two microRNAs (miR-133a-5p and miR-144-3p) were upregulated in seven patients and six microRNAs (miR-133a-3p, miR-133b, miR-206, miR-20a-5p, miR-301a-3p, and miR-32-5p) were upregulated in six patients. During the technical validation phase, we found significant upregulation of miR-144-3p expression in late-stage osteoarthritis compared with early-stage osteoarthritis. Expression of the other microRNAs was not significantly different according to the paired-t test. However, miR-133a-3p, miR-133b, and miR-206 were upregulated >2-fold in 62.5% or more of patients with late-stage osteoarthritis. Some of the microRNAs identified in this study might be involved in joint capsule degeneration or synovitis.


Lysine-specific demethylase 1 (LSD1) serves as an potential epigenetic determinant to regulate inflammatory responses in mastitis.
It is well-established that lysine-specific demethylase 1 (LSD1) is the first identified histone demethylase. Based on its demethylase enzymatic activity, LSD1 plays a pivotal role in vast range of cellular processes and cancers, but the understanding of its effects on inflammation is relatively limited. Using in vivo models of lipopolysaccharide (LPS)-induced inflammation and in vitro assays in mouse mammary epithelial cells, we identified the novel regulatory roles and underlying mechanisms of LSD1 on LPS-induced mastitis. Mammary gland and cells were collected for the following experiments after treatment. Histological changes were determined by H&E. Western blot analysis was used to detect the protein expression. ELISA and real-time PCR were used to evaluate protein and mRNA expression of inflammatory genes. Our results showed that LPS treatment resulted in a significant increase in LSD1 protein expression. GSK-LSD1 is a selective inhibitor of LSD1 enzyme activity. Treatment of mice with GSK-LSD1 inhibited LSD1 activity, reduced inflammatory cells recruitment to tissues and attenuated LPS-induced damage in mammary gland. Mechanistic investigations suggested that LSD1 inhibition led to the increase of histone H3K4me2 and H3K9me2. Furthermore, GSK-LSD1 inhibition of LSD1 further inhibited nuclear factor κ-B (NF-κB) signaling cascades, and subsequently inhibited the production of cytokines (TNF-α, IL-6 and IL-1β) in mammary gland. Taken together, our data reveal LSD1 as a potential regulator of inflammation and improve our understanding of epigenetic control on inflammation.


Ocular vestibular evoked myogenic potentials in patients with acoustic neuroma.
To assess the usefulness of vestibular testing in patients with acoustic neuroma, considering two main aspects: to compare diagnostic sensitivity of the current vestibular tests, especially considering ocular vestibular evoked myogenic potentials (OVEMPs) and to identify pre-operative localization of the tumor (inferior vestibular nerve vs. superior vestibular nerve) only with the help of vestibular electrophysiological data. Twenty-six patients with unilateral acoustic neuroma (mainly intracanalicular type) were studied with a full audio-vestibular test battery (pure tone and speech audiometry, caloric bithermal test, vibration-induced nystagmus test (VIN), cervical and OVEMPs). 18 patients (69 %) showed abnormal caloric responses. 12 patients (46.2 %) showed a pattern of VIN test suggestive of vestibular asymmetry. 16 patients (61.5 %) showed abnormal OVEMPs (12 only to AC, 4 both to AC and BC). 10 patients (38.5 %) showed abnormal cervical vestibular evoked myogenic potentials (5 both to AC and BC, 5 only to AC). In one case, results of vestibular evoked potentials and caloric test were confirmed by intra-operative and post-operative findings. Results of electrophysiological tests in AN patients could be helpful for planning the proper surgical approach, considering that sensitivity of every exam is quite low in intracanalicular lesion; clinical data allow a better interpretation of vestibular evoked myogenic potentials.


Isosteviol prevents the prolongation of action potential in hypertrophied cardiomyoctyes by regulating transient outward potassium and L-type calcium channels.
Cardiac hypertrophy is a thickening of the heart muscle that is associated with cardiovascular diseases such as hypertension and myocardial infarction. It occurs initially as an adaptive process against increased workloads and often leads to sudden arrhythmic deaths. Studies suggest that the lethal arrhythmia is attributed to hypertrophy-induced destabilization of cardiac electrical activity, especially the prolongation of the action potential. The reduced activity of Ito is demonstrated to be responsible for the ionic mechanism of prolonged action potential duration and arrhythmogeneity. Isosteviol (STV), a derivative of stevioside, plays a protective role in a variety of stress-induced cardiac diseases. Here we report effects of STV on rat ISO-induced hypertrophic cardiomyocytes. STV alleviated ISO-induced hypertrophy of cardiomyocytes by decreasing cell area of hypertrophied cardiomyocytes. STV application prevented the prolongation of action potential which was prominent in hypertrophied cells. The decrease and increase of current densities for Ito and ICaL observed in hypertrophied myocytes were both prevented by STV application. In addition, the results of qRT-PCR suggested that the changes of electrophysiological activity of Ito and ICaL are correlated to the alterations of the mRNA transcription level.


25-Hydroxycholesterol exerts both a cox-2-dependent transient proliferative effect and cox-2-independent cytotoxic effect on bovine endothelial cells in a time- and cell-type-dependent manner.
25-hydroxycholesterol (25-OHC) is a product of oxidation of dietary cholesterol present in human plasma. 25-OHC and other oxidized forms of cholesterol are implicated in modulating inflammatory responses involved in development of atherosclerosis and colon carcinogenesis. Primary lymphatic, venous and arterial endothelial cells isolated from bovine mesentery (bmLEC, bmVEC, bmAEC) were treated with 25-OHC and tested for several different cellular parameters. We found 25-OHC to be a potent inducer of cyclooxygenase-2 (Cox-2, prostaglandin G-H synthase-2) expression in bovine mesenteric lymphatic, venous, and arterial endothelial cells. The induction of Cox-2 expression in endothelial cells by 25-OHC led to an initial increase in cellular proliferation that was inhibited by the Cox-2 selective inhibitor celecoxib (Celebrex). Prolonged exposure to 25-OHC was cytotoxic. Furthermore, endothelial cells induced to express Cox-2 by 25-OHC were more sensitive to the effects of the Cox-2 selective inhibitor celecoxib (Celebrex). These results suggest that some effects of 25-OHC on cells may be dependent on Cox-2 enzymatic activity. Cox-2 dependent elevating effects of 25-OHC on endothelial cell proliferation was transient. Prolonged exposure to 25-OHC caused cell death and enhanced celecoxib-induced cell death in a cell-type dependent manner. The lack of uniform response by the three endothelial cell types examined suggests that our model system of primary cultures of bmLECs, bmVECs, and bmAECs may aid the evaluation of celecoxib in inhibiting proliferation of different types of tumour-associated endothelial cells.


Guidelines on controlling latent tuberculosis infection to support tuberculosis elimination.
Latent TB infection (LTBI) affects 25% of the world's population. As long as this reservoir exists, the elimination of TB will not be feasible. The Assembly of the World Health Organization adopted the "Global End TB" strategy for the elimination of TB in 2014. The objective of this review is to present strategies for risk groups that are candidates for the detection and treatment of LTBI. There is sufficient evidence of screening in: immunocompromised people (HIV-infected, biological therapies, alternative renal therapy, organ transplantation), recent immigrants, inmates in prison, people injecting drugs and homeless people, and workers from at-risk settings. Tests to diagnose LTBI include tuberculin skin test (TST) and gamma release assay interferon (IGRA). There is no reference test and the choice of one or the other will depend on logistical considerations, such as avoiding injection (TST) or not needing a second visit (IGRA). Treatment of LTBI is based on the use of isoniazide and rifampicin in short period of 3 or 4 months, using associations of rifampicin and isoniazide or rifampicin alone. Given the estimated high prevalence of LTBI, renewed efforts are required to reduce the number of people with LTBI that includes a registration and monitoring system to observer progress, increased testing, and the use of short treatment guidelines.


Enhancement of algicidal activity by immobilization of algicidal bacteria antagonistic to Stephanodiscus hantzschii (Bacillariophyceae).
Enhancement of algicidal activity by immobilization of algicidal bacteria antagonistic to Stephanodiscus hantzschii. In laboratory studies, A diatom-lysing bacterium, Pseudomonas fluorescens HYK0210-SK09 showed strong algicidal activity against S. hantzschii, but a natural mesocosm study revealed that this bacterium failed to fully control natural blooms of Stephanodiscus at the low water temperatures that favour these blooms. Here, we sought to develop an effective immobilization strategy for enhancing the algicidal activity of HYK0210-SK09 in the natural setting. Bacterium HYK0210-SK09 was immobilized with various carriers including agar, alginate, polyurethane and cellulose sponge. The bacterial cells immobilized with cellulose sponge (CIS) induced more rapid and complete lysis of S. hantzschii than other carriers, and had a higher packing ability than polyurethane. Furthermore, CIS-immobilized cells showed higher lysis of S. hantzschii at the same concentrations as that of free cells (< or =1 x 10(7) cells ml(-1)), and had especially strong algicidal activity at the low temperatures (<10 degrees C). Based on these laboratory studies, we assessed the possible application of HYK0210-SK09 cells in the field by performing a mesocosm study during the winter season. The CIS-immobilized cells with species-specific activity towards the genera Stephanodiscus showed extremely high algicidal activity (up to 95%) against a bloom of Stephanodiscus hantzschii even at low water temperatures, because of high cell packing and subsequent cell protection against low temperatures and predators, whereas free cells showed negligible algicidal activities under these conditions. Immobilizing cells of HYK0210-SK09 in CIS foam, rather than in the other matrices tested, could achieve more efficient control of Stephanodiscus blooms and showed a significant algicidal activity on in vitro and in vivo blooms, even at low water temperature. Collectively, these results indicate that CIS of algicidal bacteria may form an important strategy for effective management of Stephanodiscus blooms at low water temperatures.


Radiation-induced chromosomal instability and gene expression profiling: searching for clues to initiation and perpetuation.
Radiation-induced genomic instability (RIGI) manifests in the progeny of cells surviving ionizing radiation (IR), and can be measured using such endpoints as delayed mutation, micronuclei formation, and chromosomal instability. The frequency of RIGI is relatively high, exceeding the gene mutation rate of IR by orders of magnitude, leading to conjecture that a gene mutation is not the cause of the phenotype. We have started to explore whether differential gene expression patterns are associated with the instability phenotype, in order to shed light on its initiation and perpetuation. Using GM10115 human-hamster hybrid-derived chromosomally stable and radiation-induced unstable clones, gene expression patterns were analyzed using microarray analysis. Two methods were used to find differentially expressed genes, and all candidate genes identified by these methods were under-expressed relative to the chromosomally stable reference sample. Among this set differentially expressed genes identified were two candidates with a relationship to the ubiquitin/proteasome pathway. While follow-up gene expression analyses have confirmed the under-expression of these two genes in some of our chromosomally unstable clones, preliminary functional studies have been unable to demonstrate a link to instability. It is anticipated that as we apply this technology to the study of radiation-induced genomic instability, clues to its onset will be revealed, ultimately contributing to a greater understanding of the mechanisms of radiation carcinogenesis.


Predicting postoperative delirium after hip arthroplasty for elderly patients using machine learning.
Postoperative delirium (POD) is a common and severe complication in elderly hip-arthroplasty patients. This study aims to develop and validate a machine learning (ML) model that determines essential features related to POD and predicts POD for elderly hip-arthroplasty patients. The electronic record data of elderly patients who received hip-arthroplasty surgery between January 2017 and April 2021 were enrolled as the dataset. The Confusion Assessment Method (CAM) was administered to the patients during their perioperative period. The feature section method was employed as a filter to determine leading features. The classical machine learning algorithms were trained in cross-validation processing, and the model with the best performance was built in predicting the POD. Metrics of the area under the curve (AUC), accuracy (ACC), sensitivity, specificity, and F1-score were calculated to evaluate the predictive performance. 476 Arthroplasty elderly patients with general anesthesia were included in this study, and the final model combined feature selection method mutual information (MI) and linear binary classifier using logistic regression (LR) achieved an encouraging performance (AUC = 0.94, ACC = 0.88, sensitivity = 0.85, specificity = 0.90, F1-score = 0.87) on a balanced test dataset. The model could predict POD with satisfying accuracy and reveal important features of suffering POD such as age, Cystatin C, GFR, CHE, CRP, LDH, monocyte count, history of mental illness or psychotropic drug use and intraoperative blood loss. Proper preoperative interventions for these factors could reduce the incidence of POD among elderly patients.


Trends in the prevalence of peptic ulcer disease and Helicobacter pylori infection in family physician-referred uninvestigated dyspeptic patients in Hong Kong.
Peptic ulcer disease is mainly caused by Helicobacter pylori infection and the use of non-steroidal anti-inflammatory drugs. To investigate the trends in the prevalence of peptic ulcer disease, H. pylori infection and non-steroidal anti-inflammatory drug use in uninvestigated dyspeptic patients over recent years in Hong Kong. Data from consecutive patients with uninvestigated dyspeptic symptoms referred by family physicians for open access upper endoscopy during 1997 and 2003 were analysed in relation to peptic ulcer disease, H. pylori infection and non-steroidal anti-inflammatory drug use. Among 2700 patients included, 405 (15%) had peptic ulcer disease and 14 (0.5%) had gastric cancer. There was a reduced trend from 1997 to 2003 in the prevalence of peptic ulcer disease (17, 20, 14, 16, 13, 14 and 14%, respectively, chi2 = 5.80, P = 0.016) (mainly because of decrease in duodenal ulcers), H. pylori infection (44, 50, 49, 44, 40, 40, 36 and 43%, respectively, chi2 = 13.55, P < 0.001) and non-steroidal anti-inflammatory drug use (13, 5, 5, 6, 3, 4, 4 and 5% respectively, chi2 = 13.61, P < 0.001). The prevalence of peptic ulcer disease, H. pylori infection and non-steroidal anti-inflammatory drug use between 2001 and 2003 were significantly lower than that between 1997 and 2000 (17% vs. 13%, OR = 0.78, 95% CI: 0.63-0.96, P = 0.020 for peptic ulcer disease; 47% vs. 39%, OR =0.72, 95% CI: 0.60-0.86, P < 0.001 for H. pylori infection; and 6% vs. 4%, OR = 0.56, 95% CI: 0.39-0.82, P = 0.002 for non-steroidal anti-inflammatory drug use). H. pylori infection was associated with both duodenal ulcer (OR = 15.87, 95% CI: 10.60-23.76, P < 0.001) and gastric ulcer (OR = 3.12, 95% CI: 2.15-4.53, P < 0.001) whereas non-steroidal anti-inflammatory drug use was only associated with gastric ulcer (OR = 2.97, 95% CI: 1.70-5.20, P < 0.001). The prevalence of peptic ulcer disease, mainly duodenal ulcers, was reduced in association with a decreasing trend in the prevalence of H. pylori infection and non-steroidal anti-inflammatory drug use from 1997 to 2003.


The association between body esteem and sexual desire among college women.
Relationships between body image variables and sexuality have been found among several groups of women. However, research has largely focused on generalized experiences of sexuality. With the exception of two studies which focused on specific medical populations, to our knowledge there has been no investigation of the relationship between body image and acute measures of sexual response. In the current study, we investigated the relationships between body esteem, sexual response to erotica in a laboratory-setting, and self-reported sexual functioning in a non-clinical sample of 85 college women. Women participated in one study session, during which mental sexual arousal, perceptions of physical arousal, and sexual desire were assessed. Results showed that higher body esteem was significantly positively related to sexual desire in response to erotica in the laboratory setting. Similarly, higher body esteem was positively related to self-reported measures of sexual desire, as assessed by a validated measure of sexual function. The sexual attractiveness and weight concern subscales of the Body Esteem Scale, which relate to body characteristics that are most likely to be under public scrutiny, were particularly linked to sexual desire. This is the first study to show that body esteem is related to sexual responses to a standardized erotic stimulus in a laboratory setting.


Mast Cells Modulate the Immune Response and Redox Status of the Gastrointestinal Tract in Induced Venom Pathogenesis.
The pathogenesis of Androctonus autralis hector (Aah) scorpion venom involved cellular and molecular mechanisms resulting in multi-organ dysfunction. However, little is reported about the effects of venom on the gastrointestinal axis. Mast cells (MCs) are known to play a crucial role in modulating immune response of the gut. This study aims to investigate the involvement of this cell type in venom-induced gastric and intestinal disorders in a time course (3 and 24h). The obtained results revealed that Aah scorpion venom induced inflammatory cell infiltration as shown by the increase of the myeloperoxidase and eosinophil peroxidase activities. Overexpression of the c-kit receptor (CD117) severely imbalanced the redox status with depletion of antioxidant systemic accompanied by gastrointestinal tissue damage. Moreover, an increased level of lactate dehydrogenase in the serum was correlated with tissue injuries. Pharmacological inhibition of MCs targeting tyrosine kinase (TK) reduces the generation of reactive oxygen species and normalizes catalase, and gluthation S-transferase activities to their physiological levels. In addition, histopathological alterations were restored after pretreatment with c-kit receptor inhibitor associated with a considerable reduction of MC density. Interestingly, obtained results indicate that MCs might be involved in gastric modulation and intestinal inflammation through c-kit signaling following sub-cutaneous Aah venom injection.


Changes in the microbial community of an anammox consortium during adaptation to marine conditions revealed by 454 pyrosequencing.
The anammox activity of a freshwater anammox consortium was strongly inhibited at low-salinity level. Stepwise adaptation from 0 to 3 g NaCl L-1 took 153 days. Further adaptation to high-salinity concentration (from 3 to 30 g L-1) took only 40 days, and no inhibition was observed. A comprehensive insight into the salinity-induced successions of the total and the anammox communities was obtained by 454 pyrosequencing of 16S rRNA gene amplicons and statistical analysis. A major succession in the anammox community was observed at 3 g L-1 where the dominating population shifted from Candidatus Brocadia fulgida to Ca. Kuenenia stuttgartiensis. The latter dominated at high salinity and seemed to be essential for the high (˃96%) ammonium and nitrite removal efficiencies achieved. SIMPER analysis indicated that these two dominating anammox species explained most to the differences in community structure among samples and helped in identifying other important members at different salinities.


The GPR55 antagonist CID16020046 mitigates advanced glycation end products (AGEs)- induced chondrocyte activation.
Osteoarthritis (OA) is one of the most common degenerative joint diseases in aging people. The activation of chondrocytes and their dysregulation are closely related to the pathogenesis of OA. GPR55 is an unique orphan G-receptor which binds to cannabinoids. In this study, we explored the role of GPR55 in advanced glycation end productions (AGEs)- induced chondrocytes activation in cultured cells. We showed that AGEs dose dependently induced GPR55 expression in ATDC5 chondrocytes. The blockage of GPR55 by its newly discovered antagonist-CID16020046 mitigated AGEs- induced increase in cellular ROS and decrease in antioxidant NRF2. Moreover, CID16020046 showed a dose-response suppressive effect on AGEs- induced expression of the major inflammatory mediators, including COX-2 and iNOS, and the production of NO and PGE2. CID16020046 also dose responsively inhibited AGEs- induced key effectors of cartilage degradation such as MMP-3 and MMP-13. In consequence, CID16020046 showed robust inhibition on AGEs- induced type II collagen degradation. Mechanistically, our data demonstrated that CID16020046 mediated GPR55 blockage ameliorated AGEs- induced NF-κB activation as revealed by its inhibition on IκBα, nuclear p65 translocation and NF-κB promoter activity. Collectively, our study demonstrates that GPR55 signaling mediates AGEs- induced chondrocyte activation, and the targeted blockage of GPR55 pathway could be therapeutic choice in the treatment of osteoarthritis.


Dietary behavior of school-going adolescents in Bhutan: Findings from the global school-based student health survey in 2016.
Bhutan is experiencing a dual burden of undernutrition and overnutrition among adolescents. Understanding dietary behavior is vital to designing evidence-based interventions to improve adolescent nutrition and prevent non-communicable diseases in adults. The aim of this study was to assess the pattern of dietary behavior and associated sociodemographic, behavioral, and metabolic risk factors among school-going adolescents in Bhutan. The Bhutan Global School-based Student Health Survey 2016 studied students in grades 7 to 11 (N = 7576), sampled from 50 schools, randomly selected based on probability proportional to enrollment size, using a standardized self-administered questionnaire. Consumption of adequate fruits and vegetables (each at least twice daily, or a combination of at least five times daily), high-protein food at least twice weekly) in the past 30 d, no fast food in the past week, and no carbonated/sweetened drinks in the past 30 d were studied. Weighted prevalence of dietary behaviors and adjusted prevalence ratio (95% confidence interval) for factors associated with them were calculated. Of 5809 students from 13 to 17 y of age comprising 3255 (56%) girls and 3184 (54.8%) day students, 1166 (20.1%) were underweight, 1655 (28.5%) were tobacco users, and 1349 (23.2%) were alcohol users. Adequate fruit and vegetable intake, high protein consumption, not consuming fast foods and carbonated beverages were reported by 29.6%, 31.8%, 9.6%, and 14.9%, respectively. Being a day student, sex, and not reporting health risk behaviors were significantly associated with any healthy dietary behavior. Healthy eating behavior was low among Bhutanese adolescents. Policies influencing availability, affordability, and acceptability of healthy diets through peer-led, school- and community-based interventions are required to promote adolescent health and prevent non-communicable diseases.


Effects of fluoxetine withdrawal in the brainstem and hypothalamus of overnourished rats: Chronic modulation on oxidative stress levels.
Poor nutritional quality in the early stages of development is associated with neurological diseases in adulthood. Studies showed that obesity-induced oxidative stress contributes to the genesis of neurological diseases through dysregulation of the brainstem and hypothalamus. Fluoxetine (Fx) is an antidepressant member in the family of selective serotonin reuptake inhibitors (SSRI) that can induce positive effects by reducing oxidative damage in brain tissues. We aimed to evaluate the late effect of Fx in the brainstem and hypothalamus of overnourished rats during development. Male Wistar rats, after birth, were randomly divided into the normal-nourished group (N, n = 9) and the overnourished group (O, n = 3). On the 39th day of life, the groups were subdivided into normofed, and the overnourished group treated or not with fluoxetine (10 mg/kg daily) (NF, NV, OF, and OV). All groups were treated from the 39th to the 59th day of life, and within 90 days, the tissues were collected for oxidative stress analysis. Briefly, our results showed that Fx treatment induced a tissue-dependent long-lasting effect in overfed animals, increasing the enzymatic defense (i.e., CAT and GST activity) in the hypothalamus, but more intensive, increasing the non-enzymatic defense (i.e., Total Thiols and GSH levels) in the brainstem. Overall, our study suggests that serotonin modulation at the final stage of brain development causes a long-lasting impact on brain structures in overfed rats at a different mode.


American College of Cardiology Body Mass Index Counseling Quality Improvement Initiative.
Overweight/obesity, prevalent cardiovascular risk factors in children, can be associated with increased risk of adverse outcomes in children with heart disease. The American College of Cardiology (ACC) developed quality metrics including a BMI metric related to identifying and counseling overweight and obese children presenting to cardiology clinics. This metric was used for a multicenter collaborative learning Quality improvement (QI) Project through the ACC Quality Network (QNet). Our aim was to increase the percentage of children between ages 3 and 18 years presenting to cardiology clinics at participating centers with BMI > 85th percentile who received appropriate counseling. Participating centers submitted data quarterly to QNet for a sample of patients who received counseling. A Key Driver Diagram was created to help teams drive improvement. Individual centers customized interventions and participated in network-wide educational learning sessions about QI and shared experience. Statistical process control charts were used. From 04/01/2017 to 09/30/2019, 27,511 patient visits were included. Among 32 participating centers, overall counseling rate was 54%. The BMI counseling rate increased from 25% in 2017Q2 to 54% in 2019Q3. There was a wide variation from 10 to 100% in the performance of individual centers. The overall rate of identification and counseling of overweight and obese children presenting to ambulatory cardiology clinics in participating centers is low. There is wide variation in the performance of centers, providing an opportunity for improvement. Using this multicenter learning approach, individual centers have demonstrated improvement. This demonstrates that collaborative learning approaches in QI can increase implementation of the metric.


Transcriptional profiling of murine macrophages stimulated with cartilage fragments revealed a strategy for treatment of progressive osteoarthritis.
Accumulating evidence suggests that synovitis is associated with osteoarthritic process. Macrophages play principal role in development of synovitis. Our earlier study suggests that interaction between cartilage fragments and macrophages exacerbates osteoarthritic process. However, molecular mechanisms by which cartilage fragments trigger cellular responses remain to be investigated. Therefore, the current study aims at analyzing molecular response of macrophages to cartilage fragments. To this end, we analyzed the transcriptional profiling of murine macrophages exposed to cartilage fragments by RNA sequencing. A total 153 genes were differentially upregulated, and 105 genes were down-regulated in response to cartilage fragments. Bioinformatic analysis revealed that the most significantly enriched terms of the upregulated genes included scavenger receptor activity, integrin binding activity, TNF signaling, and toll-like receptor signaling. To further confirm our results, immunohistochemical staining was performed to detected regulated molecules in synovial tissues of OA patients. In consistence with RNA-seq results, MARCO, TLR2 and ITGα5 were mainly detected in the intima lining layer of synovial tissues. Moreover, blockade of TLR2 or ITGα5 but not Marco using specific antibody significantly reduced production of TNF-α in stimulated macrophages by cartilage fragments. Our data suggested that blocking TLR2 or ITGα5 might be promising therapeutic strategy for treating progressive osteoarthritis.


Activation of group-II metabotropic glutamate receptors blocks induction of long-term potentiation and depotentiation in area CA1 of the rat in vivo.
The metabotropic glutamate receptor group-II agonist (1S,3S)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3S-ACPD; 5 microliters/10 mM, i.c.v.) prevented the induction of long-term potentiation and depotentiation in the CA1 area of the hippocampus in urethane-anaesthetised rats. These effects were prevented by the group-II metabotropic glutamate receptor antagonists alpha-methyl-(2S,3S,4S)-alpha-(carboxycyclopropyl)glycine (MCCG; 5 microliters/100 mM) and (RS)-alpha-methyl-4-tetrazolylphenylglycine (MTPG; 5 microliters/500 mM). The group-I antagonist (RS)-1-aminoindan-1,5-dicarboxylic acid (AIDA; 5 microliters/200 mM) or the group-III antagonist alpha-methyl-L-2-amino-4-phosphonobutyrate (MAP4; 5 microliters/100 mM) did not affect the block of the induction of long-term potentiation by 1S,3S-ACPD. It is concluded that activation of group-II metabotropic glutamate receptors can block both high-frequency stimulation-induced long term potentiation and low-frequency stimulation-induced depotentiation in the CA1 area in vivo.


Origin of the desmoplasia in desmoplastic malignant melanoma.
Four cases of desmoplastic malignant melanoma were examined light microscopically and immunohistochemically. Electron microscopy was performed in three cases. Light microscopy showed that all tumors were composed of neoplastic spindle cells that infiltrated between mature collagen bundles in the reticular dermis. Some of the spindle cells had bizarre nuclei, whereas other spindle cells resembled normal fibroblasts. Melanin could not be demonstrated in any of the tumors by histochemical techniques. Electron microscopic examination of the spindle cells showed prominence of rough endoplasmic reticulum, which was dilated and filled with flocculent material and occasional collagen fibrils. The same cells contained aggregates of non-membrane-bound melanin granules and pre-melanosomes. Some cells also showed features of myofibroblasts. Immunoperoxidase staining with anti-S100 protein antibody demonstrated positivity of the spindle cells as well as of melanocytes in the basal layer of the epidermis. Scar tissue and fibroblasts did not stain. These findings show that the desmoplastic component of these malignant melanomas derives from melanocytes that have undergone adaptive fibroplasia. Therefore, in assessing depth of invasion in a malignant melanoma, measurements should include the desmoplastic areas.


Feasibility of laparoscopic common bile duct exploration in a rural centre.
Laparoscopic common bile duct exploration has emerged as a preferred option for the management of choledocholithiasis. The present study sought to review the feasibility of this technique in a rural centre. A comprehensive retrospective review was undertaken of all patients who underwent surgical treatment of biliary calculi in Lismore, NSW (Australia), between January 1996 and December 2002. During the study period, 1567 consecutive patients underwent laparoscopic cholecystectomy, of whom 82 (5.2%) had choledocholithiasis identified at intraoperative cholangiography. A total of 86 laparoscopic common bile duct explorations were undertaken in these patients, 37 (43%) via a transcystic approach, and 49 (57%) via a laparoscopic choledochotomy. All common bile duct calculi were successfully removed in 78 cases, representing an overall duct clearance rate of 90.7%. Complications were noted in seven patients, a morbidity rate of 8.5%. Median operative time for the procedure over the study period was 173 min. Median hospital stay was 6 days for all patients. Laparoscopic common bile duct exploration can be successfully undertaken in a rural setting by general surgeons who have appropriate laparoscopic experience, and should be the procedure of choice for the management of choledocholithiasis in these patients. It should not be restricted to specialized surgical departments in major referral centres.


One-step synthesis of a suspended ultrathin graphene oxide film: application in transmission electron microscopy.
Ultrathin graphene films find their use as advantageous support for nano- and biomaterials investigations. Thin film causes a very slight deterioration to measured signals, thus providing more details of the object's structure at nanoscale. The ultimate thinness of graphene works in the best way for this purpose. However, obtaining suspended thin film of a large-area, which is convenient for applications, is often a relatively complicated and time-consuming task. Here we present a one-step 1-min technique for synthesis of an extremely thin (about 1-2 nm) continuous film suspended over cells of a conventional copper grid (50-400 μm mesh). This technique enables us to acquire a large-area film which is water-resistant, stable in organic solvents and can act as a support when studying nanoparticles or biomaterials. Moreover, the very mechanism of the film formation can be interesting from the point of view of other applications of ultrathin graphene oxide papers.


Ruthenium(II)-Catalyzed Oxidative Double C-H Activation and Annulation Reaction: Synthesis of Indolo[2,1-a]isoquinolines.
The first metal-catalyzed double aryl C(sp2)-H bond activation of antipyrine and alkyne annulation reaction is reported. This Ru(II)-catalyzed reaction was accomplished in the presence of 20 mol % phosphine ligand tricyclohexylphosphine tetrafluoroborate to afford indolo[2,1-a]isoquinolines that are very important compounds because of their bioactivity and interesting optical properties.


L-methionine antagonism of cis-platinum nephrotoxicity.
L-Methionine administered simultaneously with cis-platinum (CDDP) iv results in a significant reduction of the nephrotoxicity normally associated with CDDP without any apparent effect on the antineoplastic activity for rats bearing the Walker 256 carcinosarcoma. CDDP given with L-methionine at a 1:20 mole ratio can be administered to rats at doses up to 35 mg/kg iv with the survival of all treated animals (3/3) and up to 56 mg/kg iv (bolus injection) with the survival of 3/6 animals, while CDDP administered alone at these levels is lethal. A reduced level of protection against the nephrotoxicity was also achieved at lower mole ratios of L-methionine to CDDP. Renal function was monitored using BUN and serum creatinine levels, and gastrointestinal toxicity by weight changes during the course of the experiments. A histopathological examination of the kidneys was also performed to evaluate the protection provided by L-methionine. Under the conditions used, the reaction between L-methionine and CDDP does not appear to proceed so rapidly as to interfere with the antitumor activity of the CDDP. The examination of structural analogs as agents for the control of CDDP-induced nephrotoxicity revealed that the C-S-C-group is the essential group for the protective action in these structures. Although L-methionine can provide renal protection in rats given high doses of CDDP, it does not prevent the accumulation of platinum in the kidney.


Seasonal variation in placental abruption.
To characterize seasonal patterns of placental abruption among Jewish and Bedouin parturients in the Southern part of Israel. A retrospective population-based study comparing all singleton pregnancies of patients with and without placental abruption was conducted. Deliveries occurred between the years 1988 and 2010. A 'classical' model of time series was used, allowing to assess trend and periodic patterns of placental abruption. During the study period, 241,408 deliveries took place, of which 1685 (0.7%) were complicated with placental abruption. Placental abruption was significantly more common among Bedouin parturients: 0.77% (n = 948) vs. 0.623% (n = 737), p < 0.001. A non-linear negative correlation was noted in the incidence of placental abruption (coefficient = -0.002) during the entire study period. Time series analysis demonstrated annual cycle frequency, seasonal cycle and weekly cycle of placental abruption. The seasonal incidence of placental abruption was higher during spring (B = 7.15) and lower during summer (reference) for both populations (Jewish and Bedouins). Weekly cycle showed significantly higher incidence on Saturday (B = 3.4) and lowest on Tuesday (B = -4.66) for both groups. The daily differences were accentuated in the Bedouin population (B = 3.7 vs. B = 2.93 in the Jewish population). Placental abruption was significantly more common in the Bedouin population. Both populations demonstrated the same annual and seasonal patterns, with higher incidence in spring and autumn.


Gallbladder contractility and gallstone formation in the Richardson Ground Squirrel.
The pathogenesis of cholesterol cholelithiasis in man is probably multifactorial and the mechanism by which gallbladder stasis occurs in gallstone patients has not been studied in detail. In the present study, time-dependent changes in gallbladder motility of the Richardson Ground Squirrels were investigated. 100 of animals were examined for their contractile responses of gallbladders to CCK-OP and Ach in vitro. Also, biochemical changes of serum and bile were investigated. There were no significant differences in motility of gallbladders to CCK-OP or Ach between control and cholesterol-fed animals. The results of this study indicate that gallbladder muscle contractility remains unchanged during the dietary induction of cholesterol gallstones in the Richardson Ground Squirrel.