Dengue fever mortality score: A novel decision rule to predict death from dengue fever.
Dengue fever (DF) is still a major challenge for public health, especially during massive outbreaks. We developed a novel prediction score to help decision making, which has not been performed till date. We conducted a retrospective case-control study to recruit all the DF patients who visited a medical center during the 2015 DF outbreak. Demographic data, vital signs, symptoms/signs, chronic comorbidities, laboratory data, and 30-day mortality rates were included in the study. Univariate analysis and multivariate logistic regression analysis were used to identify the independent mortality predictors, which further formed the components of a DF mortality (DFM) score. Bootstrapping method was used to validate the DFM score. In total, a sample of 2358 DF patients was included in this study, which also consisted of 34 deaths (1.44%). Five independent mortality predictors were identified: elderly age (≥65 years), hypotension (systolic blood pressure <90 mmHg), hemoptysis, diabetes mellitus, and chronic bedridden. After assigning each predictor a score of "1", we developed a DFM score (range: 0-5), which showed that the mortality risk ratios for scores 0, 1, 2, and ≥3 were 0.2%, 2.3%, 6.0%, and 45.5%, respectively. The area under the curve was 0.849 (95% confidence interval [CI]: 0.785-0.914), and Hosmer-Lemeshow goodness-of-fit was 0.642. Compared with score 0, the odds ratios for mortality were 12.73 (95% CI: 3.58-45.30) for score 1, 34.21 (95% CI: 9.75-119.99) for score 2, and 443.89 (95% CI: 86.06-2289.60) for score ≥3, with significant differences (all p values <0.001). The score ≥1 had a sensitivity of 91.2% for mortality and score ≥3 had a specificity of 99.7% for mortality. DFM score was a simple and easy method to help decision making, especially in the massive outbreak. Further studies in other hospitals or nations are warranted to validate this score.


Safety, tolerability, and efficacy of darunavir (TMC114) with low-dose ritonavir in treatment-experienced, hepatitis B or C co-infected patients in POWER 1 and 3.
This subanalysis examines the safety and efficacy of darunavir with low-dose ritonavir (DRV/r) in hepatitis B or C virus (HBV or HCV) co-infected patients in POWER 1 and 3 trials. POWER 1 and 3 enrolled treatment-experienced, HIV-infected patients with > or =1 primary protease inhibitor (PI) mutation and HIV-1 RNA >1,000 copies/mL. All patients received an optimized background regimen plus either control PI (almost all ritonavir boosted) or one of four DRV/r doses (POWER 1) or DRV/r 600/100 mg bid (POWER 3). Patients with active HBV or HCV co-infection who did not require treatment for hepatitis were included. Safety parameters were evaluated. Of 634 DRV/r and 63 control (97% ritonavir boosted) patients assessed, 13% and 16%, respectively, had active co-infection. In both groups, more patients with active co-infection than without co-infection had liver-related adverse events (AEs). These AEs were mainly asymptomatic liver transaminase elevations, although changes were slightly less in the DRV/r group (DRV/r, 13% vs. 8%; control PI, 20% vs. 12%). Only two patients (one per treatment arm) discontinued therapy due to grade 3 or 4 alanine and aspartate transaminase elevations. DRV/r was generally well tolerated in treatment-experienced, HBV or HCV co-infected patients. No differences in liver-related AEs were observed between treatment groups.


Polymorphism of Fas and Fas ligand in preterm premature rupture of membranes in singleton pregnancies.
Fetal inflammatory response has been previously shown to be involved in the pathogenesis of preterm premature rupture of membranes. We investigated the association between polymorphisms at position -670 in the Fas gene and position -124 in the Fas ligand gene demonstrated in neonatal oral mucosa cells and preterm premature rupture of membranes. Thirty-six singleton pregnancies were studied. Eighteen pregnancies were complicated by preterm premature rupture of membranes, and 18 were delivered at term without preterm premature rupture of membranes. Buccal swabs were obtained from each neonate, and extracted deoxyribonucleic acid was analyzed by polymerase chain reaction-based restriction fragment length polymorphism for an adenine (A) to guanine (G) substitution at position -670 in the Fas promoter gene and at position -124 in the Fas ligand gene. chi2 and Fisher's exact tests were used for statistical analysis. There was no difference with respect to race, maternal age, and parity between the 2 groups. Frequencies of Fas -670 AG, -AA, and -GG genotypes in preterm premature rupture of membranes group were significantly different from those in the control group (P = .004). The frequency of the heterozygous AG genotype was significantly higher in preterm premature rupture of membranes group as compared with controls (83.3% versus 33.3%, P = .003). Fas -670 AA genotype was not observed among patients with preterm premature rupture of membranes. Similarly, the difference of frequencies of the Fas ligand -124 AG, -AA, and -GG genotypes among preterm premature rupture of membranes group and controls was observed but did not reach statistical significance. Neither of the groups demonstrated homozygous GG genotype at position -124 of the Fas ligand gene. Our data indicate an association between preterm premature rupture of membranes and increased prevalence of neonatal AG genotype at -670 Fas promoter gene. Genetically predetermined regulation of the Fas/Fas ligand system appears to play an important role in the pathogenesis of the preterm premature rupture of membranes.


Effects of metformin on early pregnancy loss in the polycystic ovary syndrome.
Polycystic ovary syndrome is the most common form of female infertility in the United States. In addition to poor conception rates, pregnancy loss rates are high (30-50%) during the first trimester. We hypothesized that hyperinsulinemic insulin resistance contributes to early pregnancy loss in the syndrome, and that decreasing hyperinsulinemic insulin resistance with metformin during pregnancy would reduce the rate of early pregnancy loss. We conducted a retrospective study of all women with polycystic ovary syndrome who were seen in an academic endocrinology clinic within the past 4.5 yr and who became pregnant during that time. Sixty-five women received metformin during pregnancy (metformin group) and 31women did not (control group). The early pregnancy loss rate in the metformin group was 8.8% (6 of 68 pregnancies), as compared with 41.9% (13 of 31 pregnancies) in the control group (P < 0.001). In the subset of women in each group with a prior history of miscarriage, the early pregnancy loss rate was 11.1% (4 of 36 pregnancies) in the metformin group, as compared with 58.3% (7 of 12 pregnancies) in the control group (P = 0.002). Metformin administration during pregnancy reduces first-trimester pregnancy loss in women with the polycystic ovary syndrome.


From cellular receptors to transduction-transcription pathways for cytokines: at which level should the inhibition be targeted in inflammation?
An imbalance in cytokine homeostasis is considered to play a major part in the pathogenesis of immuno-inflammatory diseases. Since the identification and cloning of cytokines and their receptors, therapeutic approaches have been developed with the purpose of impeding the interaction between the ligand (cytokine) and its specific receptor, or interactions that involve the use of anti-inflammatory cytokines to switch off inflammation. Although some diseases have been treated successfully with cytokines or anticytokines (i.e., anti-TNF, and to a lesser extent recombinant IL-1 receptor antagonist, in rheumatoid arthritis; IFN-beta in multiple sclerosis), the fact remains that these therapies do not abrogate the concomitant use of steroids or immunosuppressive drugs, and that a significant percentage of patients do not respond to such therapies; these are important limitations. The identification of signalling pathways preferentially used in inflammatory conditions has boosted approaches that target these intracellular mechanisms. This review examines the different therapeutic approaches that may be considered for the treatment of immuno-inflammatory diseases, and discusses the advantages and disadvantages of targeting extracellular or intracellular mechanisms.


Idiotypic T cells specific for Morbillivirus nucleocapsid protein process and present their TCR to cognate anti-idiotypic CD8+ T cells.
CD8(+) T cells are activated by the presentation of antigenic peptide through MHC class I molecules. Newly synthesized proteins formed as defective ribosomal products (DRiPs) can act as a major source of antigenic peptides for MHC class I presentation pathway. Majority of these peptides are generated from the intracellular degradation of self antigens. In the present study, we have shown that newly synthesized T cell receptor (TCR) beta chains formed as DRiPs in T cells are ubiquitinated and degraded by the proteasomes. These TCR-DRiPs are processed and presented by activated T cells to cognate anti-idiotypic CD8(+) T cells. Presentation of TCR idiopeptide (peptide derived from the variable region of idiotypic TCR) by activated T cells leads to Bcl-2 expression and cytokine secretion by anti-idiotypic CD8(+) T cells. Presentation of intracellular antigen by T cells may have important implications in immunoregulation, control of lymphotropic virus infection and autoimmune diseases.


Tegaderm CHG IV Securement Dressing for Central Venous and Arterial Catheter Insertion Sites: A NICE Medical Technology Guidance.
Catheters are widely used for vascular access and for the administration of drugs or fluids in critically ill patients. This exposes patients to an infection risk. Tegaderm chlorhexidine gluconate (CHG) (developed by 3M)-a transparent securement dressing-covers and protects catheter sites and secures devices to the skin. It comprises a transparent adhesive dressing to act as a barrier against external contamination and an integrated gel pad containing an antiseptic agent. The Medical Technologies Advisory Committee (MTAC) at the National Institute for Health and Care Excellence (NICE) selected Tegaderm CHG for evaluation. One study was identified by the sponsor as relevant to the decision problem. From this, the sponsor concluded that compared with standard dressings, Tegaderm CHG is associated with lower rates of catheter-related infection, but increased dermatitis incidence. The External Assessment Centre (EAC) identified four paired comparative studies between Tegaderm CHG, other CHG dressings or standard dressings. The EAC agreed with the sponsor's conclusion, finding that CHG dressings reduce infections compared with standard dressings. The sponsor constructed a de novo costing model. Tegaderm CHG generated cost savings of £77.26 per patient compared with standard dressings and was cost saving in 98.5 % of a sample of sets of inputs (2013 prices). The EAC critiqued and updated the model's inputs, yielding similar results to those the sponsor estimate. The MTAC reviewed the evidence and decided to support the case for adoption, issuing a positive draft recommendation. After a public consultation, NICE published this as Medical Technology Guidance 25.


Effects of environment, seedling age, and seed source on leaf resistance of three species of Chamaecyparis and Tsuga chinensis.
The objectives of this study were to determine how stomata of Chamaecyparis spp. react to environmental changes and to determine in what ways leaf resistance patterns may have adaptive value for these confers. Leaf resistances were measured of Chamaecyparis taiwanensis, C. formosensis and C. lawsoniana seedlings in a forest nursery, and of saplings of the first two species and Tsuga chinensis growing in natural forest and clearcuts. Light, temperature, humidity, and sometimes xylem pressure potential were measured concurrently.In the nursery, C. lawsoniana, the species from the driest climate, had the lowest resistance, daily means for well-watered yearold plants being 2-5 s cm-1. Leaf resistances of well-watered C. formosensis and C. taiwanensis from more humid habitats were two and three times greater. Leaf resistance was more sensitive to environment in those nursery populations of C. formosensis from habitats where a dry season is more severe. There were no clear patterns of variation within the other species. Resistance of scale leaves increased with seedling size and age. Primary leaves of C. formosensis had higher average resistance than its scale leaves, and were more responsive to changes in the environment.In the field, C. taiwanensis had higher resistance than C. formosensis. Leaf resistance of Tsuga chinesis was quite similar to that of C. formosensis. Resistance decreased from winter to spring except during a severe dry spell in March.Changes in leaf resistance were most consistently correlated with humidity, which often increased during the day as humid air rose from the lowlands. Stomata opened as humidity rose even when plant xylem pressure potentials were -15 to -23 bars. Stomatal reaction of Chamaecyparis spp. varies in ways apparently adaptive to their environment.


The Microbiome in Osteoarthritis: a Narrative Review of Recent Human and Animal Model Literature.
The microbiome has recently emerged as a powerful contributor to health and illness in chronic, systemic disorders. Furthermore, new microbiome niches beyond traditional gut locations are frequently being described. Over the past 5 years, numerous pivotal studies have demonstrated associations between changes in various microbiome niches and the development of osteoarthritis (OA). Herein, we review the most impactful recent literature, including microbiome associations with disease and the potential therapeutic value of microbiome manipulation. The gut microbiome of human OA patients is enriched in specific bacterial clades, most notably Streptococcus, which correlates with OA pain, Firmicutes, and others. Most studies have focused on knee OA, although one publication demonstrated positive associations with 3 gut microbiome clades in hand OA. OA can be easily distinguished from RA by evaluating differences in oral microbiome composition. Most studies have also demonstrated a reduction in richness of the gut microbiome (alpha diversity) associated with OA. Several studies have identified bacterial signatures within human knee and hip cartilage, synovial fluid, and synovial tissue and have described changes in these patterns occurring with the development of OA. In animal models of OA, high-fat diet-induced obesity has been the most well-studied OA risk factor associated with changes in the microbiome, with numerous bacterial clades changed within the gut microbiome and associated with OA. Also in animal models, various oral supplementations, including dietary fiber, probiotics including Lactobacillus species, and cecal microbiome transplantation have all shown improvements in OA histopathology or cartilage healing. Microbiome changes are strongly associated with the OA disease process and with individual OA risk factors related to both the gut microbiome and the microbial DNA patterns in the joint. Microbiome-directed interventions have the potential to prevent or reduce the progression of OA. Future studies should investigate the mechanistic underpinnings of these microbiome associations and further define the therapeutic potential of microbiome augmentation.


Biological analysis and imaging applications of CdSe/CdSxSe1-x/CdS core-shell magic-sized quantum dot.
Although colloidal magic-sized quantum dots present great promise for biological applications due to their high stability and strong luminescence, nanotoxicological analyses are scarcely reported and biomedical applications have not been demonstrated. This is the first report on biological effects of CdSe/CdSxSe1-x/CdS core-shell magic-sized quantum dot (CS-MSQD) with specific application in breast cancer cell detection. The 2-nm CS-MSQD presents a broad bandwidth emission from 450 to 750nm, low toxicity, non-immunogenicity and biocompatibility. The CS-MSQD was conjugated to a breast cancer-specific Fab antibody, and passively diffused into cells for in vitro detection of a breast cancer cell line, demonstrating to be an unprecedented tool for biomedical applications.


Long-term outcomes of carotid endarterectomy vs transfemoral carotid stenting in a Medicare-matched database.
Carotid endarterectomy (CEA) is associated with lower risk of perioperative stroke compared with transfemoral carotid artery stenting (TFCAS) in the treatment of carotid artery stenosis. However, there is discrepancy in data regarding long-term outcomes. We aimed to compare long-term outcomes of CEA vs TFCAS using the Medicare-matched Vascular Quality Initiative Vascular Implant Surveillance and Interventional Outcomes Network database. We assessed patients undergoing first-time CEA or TFCAS in Vascular Quality Initiative Vascular-Vascular Implant Surveillance and Interventional Outcomes Network from January 2003 to December 2018. Patients with prior history of carotid revascularization, nontransfemoral stenting, stenting performed without distal embolic protection, multiple or nonatherosclerotic lesions, or concomitant procedures were excluded. The primary outcome of interest was all-cause mortality, any stroke, and a combined end point of death or stroke. We additionally performed propensity score matching and stratification based on symptomatic status. A total of 80,146 carotid revascularizations were performed, of which 72,615 were CEA and 7531 were TFCAS. CEA was associated with significantly lower risk of death (57.8% vs 70.4%, adjusted hazard ratio [aHR], 0.46; 95% confidence interval [CI], 0.41-0.52; P < .001), stroke (21.3% vs 26.6%; aHR, 0.63; 95% CI, 0.57-0.69; P < .001) and combined end point of death and stroke (65.3% vs 76.5%; HR, 0.49; 95% CI, 0.44-0.55; P < .001) at 10 years. These findings were reflected in the propensity-matched cohort (combined end point: 34.6% vs 46.8%; HR, 0.53; 95% CI, 0.46-0.62) at 4 years, as well as stratified analyses of combined end point by symptomatic status (asymptomatic: 63.2% vs 74.9%; HR, 0.49; 95% CI, 0.43-0.58; P < .001; symptomatic: 69.9% vs 78.3%; HR, 0.51; 95% CI, 0.45-0.59; P < .001) at 10 years. In this analysis of North American real-world data, CEA was associated with greater long-term survival and fewer strokes compared with TFCAS. These findings support the continued use of CEA as the first-line revascularization procedure.


Successful Endovascular Repair of a Kommerell's Diverticulum and a Right-Sided Aortic Arch.
A 57-year-old woman was diagnosed with Kommerell's diverticulum in the setting of a right aortic arch on computed tomography. Although asymptomatic, the maximum diameter of the aneurysm was 55 mm; thus, she underwent surgery to prevent rupture of the aneurysm. A bypass was constructed from the left common carotid artery to the left subclavian artery. A stent-graft was deployed from the distal right subclavian artery, and coil embolization of the diverticulum was performed via the left subclavian artery. She was discharged after 12 days of surgery. The postoperative four-month follow up showed a smaller aneurysm. Thoracic endovascular aortic repair is feasible and effective for Kommerell's diverticulum.


Nerve terminals in the tumor microenvironment as targets for local infiltration analgesia.
Nerve terminals within the tumor microenvironment as potential pain-mitigating targets for local infiltration analgesia is relatively less explored. In this study, we examine the role of key analgesics administered as local infiltration analgesia in a model of cancer-induced bone pain (CIBP). CIBP was induced by administration of allogenic MRMT1 breast cancer cells in the proximal tibia of rats, and tumor mass characterized using radiogram, micro-CT, and histological analysis. In vitro responsiveness to key analgesics δ-opioid receptor agonist (DOPr), Ca2+ channel and TRPV1 antagonists was assessed using ratiometric Ca2+ imaging in sensory neurons innervating the tumor site. Effectiveness of locally infiltrated analgesics administered independently or in combination was assessed by quantifying evoked limb withdrawal thresholds at two distinct sites for up to 14 days. CIBP animals demonstrated DOPr, N-, and L-type and TRPV1 expression in lumbar dorsal root ganglion neurons (DRG), comparable to controls. Evoked Ca2+ transients in DRG neurons from CIBP animals were significantly reduced in response to treatment with compounds targeting DOPr, N-, L-type Ca2+ channels and TRPV1 proteins. Behaviourally, evoked hyperalgesia at the tumor site was strongly mitigated by peritumoral injection of the DOPr agonist and T-type calcium antagonist, via its activity on bone afferents. Results from this study suggest that nerve terminals at tumor site could be utilized as targets for specific analgesics, using local infiltration analgesia.


New concept for a toxicity assay based on multiple indexes from the wave shape of damped metabolic oscillation induced in living yeast cells (part I): characterization of the phenomenon.
The damped glycolytic oscillation phenomenon occurring in starved cells of the yeast Saccharomyces cerevisiae (NBRC 0565) was characterization for application to a toxicity bioassay. S. cerevisiae was grown under semi-anaerobic conditions. The transient oscillations were observed photometrically as the time course of the fluorescent intensity of reduced pyridine nucleotide resulting from instantaneous addition of glucose to a cell suspension. In this study, simple and reproducible conditions inducing damped oscillations were obtained by modifying a literature method. For estimation of the wave shapes of the damped oscillations we used six indexes. To investigate the total reproducibility as the averaged relative standard deviation (RSD(av)) for the six indexes obtained from the wave shapes, the damped oscillations were induced under the optimum conditions and the RSD(av) values were calculated as 14% in a buffer cell suspension (n = 62) and 22% in a water cell suspension (n = 78). Finally, the effects of glucose concentration on the six indexes were examined, and all the indexes changed when the glucose concentration was changed. Excellent correlations were obtained between the index of oscillation-state time and the concentration of glucose in a buffer cell suspension (r = 0.9985, 0.5-250 mmol L(-1), 10 points) and in a water cell suspension (r = 0.9989, 2.5 micromol L(-1)-250 mmol L(-1), 12 points), respectively.


Radical host-specific therapies for TB.
Although proinflammatory cytokines such as TNF are critical for containment of tuberculosis, they can also exacerbate disease when produced at high levels. In this issue of Cell, Roca and Ramakrishnan demonstrate that high TNF production induces reactive oxygen species in infected macrophages, ultimately leading to macrophage necrosis and bacterial dissemination.


The immune response of peritoneal macrophages due to exposure to inorganic lead in the house mouse Mus musculus.
Peritoneal macrophages from the house mouse (Mus musculus) were exposed to variable lead (Pb) concentrations (0.2, 2, 20 and 40 microM) to better understand lead cytotoxicity and its damage to the immune response. Phagocytes were exposed to 20 and 40 microM Pb for 72 h, and macrophages were exposed at lower concentrations (0.2, 2 and 20 microM Pb) for 24h and 72 h. Dysfunctions in macrophage immune activity were examined by measuring phagocytic activity, nitric oxide production, endosomal/lysosomal stability and cell adhesion. Lead affected all macrophage functions, even at low concentrations, by reducing the phagocytic index, nitric oxide production, endosomal/lysosomal system stability and cell adhesion, and upregulating the antioxidant enzymatic activity of catalase. We demonstrate that lead affects the redox status of the cells and suggest that the immunomodulatory effects at low dosages on mouse macrophages reduces their ability to protect the host against infectious agents.


Differences in iron deficiency anemia and mean platelet volume between children with simple and complex febrile seizures.
The relationship between iron deficiency anemia and febrile seizures (FSs) were examined in several studies before. The aim of our study is to find out the differences regarding iron deficiency anemia, demographic characteristics and mean platelet volume (MPV) which is an inflammatory marker between simple and complex febrile seizure groups. In this study, the authors investigated the recordings of 493 children with a diagnosis of simple and complex febrile seizure, aged between 6 months and 6 years, followed between 2002 and 2010 retrospectively. Mean age and male/female ratio were similar in two groups. There was no significant difference regarding with age, gender and family history of FS between two groups. We found significant difference statistically with respect to gestational age, consanguinity, family history of epilepsy and birth weight between two groups. The mean levels of Hb, Htc, MCV were lower and Plt and RDW levels were higher in children with CFS than SFS group, the differences were statistically significant (p: 0.001). A higher proportion of children with CFS (16.2%) had iron deficiency anemia compared to SFS group (12.1%). Mean platelet volume (MPV) of CFS (7.99±0.96fL) were significantly lower than that of SFS group (8.77±0.75) (p<0.001). The results of this study suggests that iron deficiency anemia is more frequently seen among the patients with CFS than the patients with SFS. The lower levels of MPV as an inflammatory marker, supports the idea that CFS is a brain inflammatory disease and the consequence of this inflammatory mechanism is the development of the epilepsy. Further studies are necessary to highlight the relationship between iron metabolism, inflammation and seizures.


Mutant p53 expression enhances drug resistance in a hepatocellular carcinoma cell line.
Chemoresistance is a major problem in the treatment of hepatocellular carcinoma. Certain p53 mutants may enhance drug resistance in cancer cells. To determine whether two frequently occurring p53 mutants, R248Q and R273C, would increase the drug resistance of liver cancer cells, stable cell lines expressing these specific p53 mutants were established by transfecting the p53-null Hep3B cells with mutant p53 expression vectors, and then treating them with the anticancer drugs doxorubicin and paclitaxel. The cells expressing the p53 mutant, R248Q, but not R273C, displayed cross-resistance to both drugs, in contrast to the control cells expressing the vector alone. Moreover, both the expression and the activity of the multiple drug resistance gene product, P-glycoprotein, were elevated in p53 mutant R248Q-expressing cells. Reduced uptake of doxorubicin was also observed in the R248Q-expressing cells. These results suggest that expression of the p53 mutant, R248Q, in liver cancer cells may enhance their drug resistance and that upregulation of P-glycoprotein activity may contribute to this protective effect.


[Emergency mission documentation in simulated care. Video-based error analysis].
The aim of this study was to examine documentation quality in physician staffed emergency medical services (EMS). This study compared simulated on-site care with the associated patient records written by EMS physicians. For this study two standardized simulated case scenarios, ST segment elevation myocardial infarction (STEMI) and major trauma with traumatic brain injury were designed by an expert committee. Overall 29 EMS teams each consisting of 1 EMS physician and 2 paramedics ran through the scenarios on high fidelity patient simulators and each scenario was videotaped. The scenarios were stopped after 12 min for STEMI and after 14 min for major trauma independent of the actions carried out and each EMS physician then had 10 min to document this initial phase on standardized protocol sheets. The videotaped scenarios were analyzed by two independent investigators. Documentation of predefined contents and all drug dosages were checked against the simulated on-site care. The data were evaluated and classified as correct, incorrect or missing documentation although action performed. Written consent for data analysis was provided by 28 teams. Overall 20 parameters and actions in the STEMI scenario and 16 in the major trauma scenario as well as all drug dosages were evaluated. For the scenario STEMI 469 actions were analyzed of which 271 (58%) were correct, 94 (20%) incorrect and 104 (22%) had missing documentation. A total of 140 medications were administered of which 31 (22%) were documented incorrectly and 14 (10%) were not documented. For major trauma 401 actions were analyzed of which 244 (61%) were correct, 101 (25%) incorrect and 56 (14%) had missing documentation. In this scenario the teams administered 138 medications of which 31 (22%) were documented incorrectly and 16 (12%) were not documented. Infused amounts of crystalloids and colloids were mostly documented correctly in this case (35 correct /6 incorrect/8 not documented). Documentation of several clinical parameters was carried out predominantly correctly, e.g. initial blood pressure (STEMI: 25/2/1, major trauma: 21/4/2) and initial ECG rhythm (STEMI: 27/0/1, major trauma: 26/0/1). Documentation of other clinically relevant parameters was often performed incorrectly: 12-lead ECG in STEMI (5/9/12) and capnometry in major trauma (9/4/7). No team used a pain scale to assess the level of pain in the STEMI scenario but 12 EMS physicians documented an accordant value (numerical rating scale) on the patient records. Furthermore some parameters could be identified where documentation was mostly missing although they were measured, e.g. onset of symptoms in STEMI (5/4/15) and reduced level of consciousness and bradypnea in major trauma (9/2/17). Patient safety can be reduced if relevant preclinical data are not transmitted correctly to the admitting hospital. Therefore there is a need to improve documentation quality in EMS. Electronic documentation, training of EMS staff and quality management programs might offer solutions. Because of the small sample size further studies are needed to evaluate the validity of these results.


A new ambulatory monitoring system for tremor.
We have described a fully portable tremor recording system which allows a patient to carry out his normal day's activities with minimal hindrance. Computer analysis of the recording has enabled the extent of the tremor over prolonged periods to be presented in a format which is easily understandable and which should be useful for the objective assessment of therapy.


Eruptive Xanthomas: Importance of Recognition to Reduce Delay of Effective Triglyceride Reduction.
Eruptive xanthomas are localized lipid deposits in the dermis and an important early clue to severe hypertriglyceridemia. These small erythematous or yellow papules that localize to the extensor surfaces of extremities, buttocks, and the back are often overlooked during routine visits secondary to poor familiarity and limited skin examinations. We present 3 cases of patients with eruptive xanthomas and severe hypertriglyceridemia who underwent skin biopsy and waited weeks to years before receiving effective treatment. We suggest the following to minimize the delay between presentation and effective management. First, perform a comprehensive skin examination. Second, be mindful of the association between metabolic syndrome or diabetes with severe hypertriglyceridemia. Third, evaluate the Four D's of secondary hypertriglyceridemia: Diet/Lifestyle, Drugs/Medications, and Diseases/Disorders of metabolism. Finally, initiate effective treatment promptly after recognition. This includes beginning with a minimal fat diet and appropriate pharmacological intervention to control triglycerides as outlined in recent guidelines.


Design and synthesis of phosphinamide-based hydroxamic acids as inhibitors of matrix metalloproteinases.
A new series of hydroxamic acid-based matrix metalloproteinase (MMP) inhibitors containing a unique phosphinamide motif derived from D-amino acid was designed, synthesized, and tested for enzyme inhibition. Compounds with an R configuration at phosphorus were found to be potent MMP inhibitors while molecules with the S configuration were almost inactive. Structure-activity relationship studies of the series led to the discovery of the potent inhibitor 16 with IC50 = 20.5 nM and 24.4 nM against fibroblast collagenase (MMP-1) and stromelysin (MMP-3), respectively. The binding mode of this novel phosphinamide-based series of MMP inhibitors was established based on X-ray crystallography of the complex of stromelysin and 16.


Enhancing effects of Ro 15-1788 on straw-climbing behavior as measured with the straw-suspension method: reversal by diazepam.
Ro 15-1788 (0.5, 1, 5, or 20 mg/kg), a pure benzodiazepine receptor antagonist, was studied for its effect on the duration of immobility and the number of straw-climbing attempts in a modified forced-swim test with straw-suspension of rats. A single dose of 20 mg/kg of Ro 15-1788 injected IP prolonged only the duration of immobility with no effect on straw-climbing behavior, whereas both doses of 1 and 5 mg/kg of this compound significantly enhanced the number of straw-climbing attempts in an inverted U-shaped manner. Ro 15-1788 at 1 mg/kg significantly reversed the inhibitory effect of 1 mg/kg of diazepam on the number of straw-climbing attempts. It is suggested that the enhancing effect of low doses of Ro 15-1788 on straw-climbing behavior can be regarded as an index of its anxiogenic effect, by acting via central benzodiazepine receptors.


Do mitochondria contribute to left ventricular non-compaction cardiomyopathy? New findings from myocardium of patients with left ventricular non-compaction cardiomyopathy.
Left ventricular non-compaction cardiomyopathy (LVNC) is a rare congenital cardiomyopathy that is associated with mutations in mitochondrial DNA (mtDNA), however, no study of myocardium mtDNA of LVNC patients has been reported. To identify novel candidate mtDNA variants that may be responsible for the pathogenesis of LVNC, myocardial specimens were examined to investigate pathogenic mtDNA variants. Samples from six patients who were diagnosed with LVNC and underwent heart transplantation were analyzed. The sequence and copy number of mtDNA from these samples were determined by Sanger sequencing and fluorescence-based quantitative polymerase chain reaction, respectively. Myocardial mtDNA sequences analysis revealed 227 substitution variants, including 157 coding variants and 70 non-coding variants. An m.9856T>C (Ile217Thr) mutation in MT-CO3 from one LVNC patient was found to be a non-haplogroup associated variant, and was rare in the mtDB Human Mitochondrial Genome Database, suggesting that the variant may be pathogenic. And there was statistically significant difference in mtDNA copy number between LVNC patients and normal control subjects. Electron microscopy (EM) of left ventricular myocardium showed abnormality in mitochondrial morphology and disordered sarcomeric organization. The identification of mtDNA sequence variants in myocardial specimens may be helpful for further investigation of the underlying pathogenic implications of myocardial mtDNA mutations in LVNC. However, measurement of mtDNA copy number showed that there was lower mtDNA content in myocardium of LVNC patients than in normal controls (P<0.01). Lower mtDNA copy number and morphological abnormalities of mitochondria suggested mitochondrial dysfunction that may be associated with etiology of LVNC.


In-line micro-matrix solid-phase dispersion extraction for simultaneous separation and extraction of Sudan dyes in different spices.
A novel, simple, fast, and miniaturized method, termed in-line micro-matrix solid-phase dispersion (in-line MMSPD), coupled with high performance liquid chromatography (HPLC) was developed for the simultaneous extraction and determination of Sudan dyes (i.e. Sudan I-IV, Sudan orange G, Sudan black B, and Sudan red G) with the aid of an experimental design strategy. In this method, a matrix solid-phase dispersion (MSPD) column including a suitable mixture of polar sorbents was inserted in the mobile phase pathway, and while the interfering compounds were retained, the analytes were eluted and entered into the analytical column. In this way, the extraction, elution, and separation of the analytes were performed sequentially. Under the optimal experimental conditions (including the amount of sample, 0.0426g; amount of dispersant phase, 0.0216g of florisil, 0.0227g of silica, 0.0141g of alumina; and blending time, 112s), the limits of detection (LODs), limits of quantification, linear dynamic ranges, and recoveries were obtained to be 0.3-15.3μgkg(-1), 1-50μgkg(-1), 50-28,000μgkg(-1), and 94.5-99.1%, respectively. The results obtained showed that determination of the selected Sudan dyes in food samples using an enough sensitive and a simple analytically validated method like in-line MMSPD may offer a suitable screening method, which could be useful for food analysis and adulteration.


Applying Stirred Perfusion to 3D Tissue Equivalents to Mimic the Dynamic In Vivo Microenvironment.
Complex three-dimensional (3D) tissue equivalents have been widely developed with applications with a multitude of organs and tissues. While these systems lead to significant improvements over conventional two-dimensional culture, the static conditions of the surrounding medium still present a limitation to the physiological relevance of these models. Medium perfusion and convective mixing can be introduced to these models through a variety of techniques using equipment such as pumps and rockers. These systems can easily become very complex or suffer from limited control over the fluid flow properties. We have developed a bioreactor enabling controlled perfusion of 3D tissue equivalents utilizing a magnetic stirrer-based system, allowing for scalability and ease of use. This system has demonstrated potential applications in a range of tissues such as the liver, intestine, and skin, with many other potential applications yet to be tested. Our solution provides users with a low cost and easy to use alternative to complex bioreactor systems while still providing high levels of control over fluid flow and structural properties of the tissue constructs.


Silver catalyzed intramolecular cyclization for synthesis of 3-alkylideneoxindoles via C-H functionalization.
A novel protocol for the preparation of various 3-alkylideneoxindoles via a silver-catalyzed aromatic C-H functionalization has been developed. The process is simple, environmentally conscious, and avoids the use of abundant bases, oxidants, or other additives.


Chloroquine and hydroxychloroquine as ACE2 blockers to inhibit viropexis of 2019-nCoV Spike pseudotyped virus.
The novel coronavirus disease (2019-nCoV) has been affecting global health since the end of 2019 and there is no sign that the epidemic is abating . The major issue for controlling the infectious is lacking efficient prevention and therapeutic approaches. Chloroquine (CQ) and Hydroxychloroquine (HCQ) have been reported to treat the disease, but the underlying mechanism remains controversial. The objective of this study is to investigate whether CQ and HCQ could be ACE2 blockers and used to inhibit 2019-nCoV virus infection. In our study, we used CCK-8 staining, flow cytometry and immunofluorescent staining to evaluate the toxicity and autophagy of CQ and HCQ, respectively, on ACE2 high-expressing HEK293T cells (ACE2h cells). We further analyzed the binding character of CQ and HCQ to ACE2 by molecular docking and surface plasmon resonance (SPR) assays, 2019-nCoV spike pseudotyped virus was also used to observe the viropexis effect of CQ and HCQ in ACE2h cells. Results showed that HCQ is slightly more toxic to ACE2h cells than CQ. Both CQ and HCQ could bind to ACE2 with KD = (7.31 ± 0.62)e-7 M and (4.82 ± 0.87)e-7 M, respectively. They exhibit equivalent suppression effect for the entrance of 2019-nCoV spike pseudotyped virus into ACE2h cells. CQ and HCQ both inhibit the entrance 2019-nCoV into cells by blocking the binding of the virus with ACE2. Our findings provide novel insights into the molecular mechanism of CQ and HCQ treatment effect on virus infection.


Influence of the Increasing Number of Organic Radicals on the Structural, Magnetic, and Electrochemical Properties of the Copper(II)-Dioxothiadiazole Family of Complexes.
The preparation, structures, and electrochemical and magnetic properties supported by density functional theory (DFT) calculations of three new copper(II) compounds with [1,2,5]thiadiazolo[3,4-f][1,10]phenanthroline 1,1-dioxide (td) and its radical anion (td·-) are reported: {[CuIICl(td)](μ-Cl)2[CuIICl(td)]} (1), which incorporates only neutral td ligands; [CuIICl(td·-)(td)]·2MeCN (2), which comprises one neutral td and one radical td·-; and PPN[CuIICl(td·-)2]·2DMA (3), where CuII ions are coordinated by two radical anions td·- (DMA, dimethylacetamide; PPN+, the bis(triphenylphosphine)iminium cation). All three compounds show interesting paramagnetic behavior with low-temperature features indicating significant antiferromagnetic coupling. The magnetic properties of 1 are dominated by CuII···CuII interactions (JCuCu) mediated through the Cl- bridges, while the magnetic properties of 2 and 3 are governed mainly by the td·-···td·- (Jtdtd) and CuII-td·- (JCutd) exchange interactions. The structure of 2 features only two major magnetic coupling pathways enabling the fitting of experimental data with Jtdtd = -36.0(5) cm-1 and JCutd = -12.6(2) cm-1 only. Compound 3 exhibits a complex network of magnetic contacts. Attempt to approximate its magnetic behavior using only a local magnetic contacts model resulted in Jtdtd = -5.6(1) cm-1 and two JCutd constants, -12.4(2) and -22.6(4) cm-1. The experimental fitting is critically compared with the results of broken symmetry density functional theory (BS DFT) calculations for inter- and intramolecular contacts. More consistent results were obtained with the M06 functional as opposed to popular B3LYP, which encountered problems reproducing some of the experimental intermolecular exchange interactions. Electrochemical measurements of 2 and 3 in MeCN showed three reversible nearly overlapping redox peaks appearing in a narrow potential range of -600 to -100 mV vs Fc/Fc+. Small differences between the redox events suggest that such compounds may be good candidates for new switchable materials, where the electron transfer between the metal and the ligand center is triggered by temperature, pressure, or light (valence tautomerism).


Propofol plus low-dose dexmedetomidine infusion and postoperative delirium in older patients undergoing cardiac surgery.
Postoperative delirium (POD) is a frequent complication in older patients. Dexmedetomidine might be effective in decreasing the incidence of POD. We hypothesised that adding low-dose rate dexmedetomidine infusion to a propofol sedation regimen would have fewer side-effects and would counteract the possible delirium producing properties of propofol, resulting in a lower risk of POD than propofol with placebo. In this double-blind placebo-controlled trial, patients ≥60 yr old undergoing on-pump cardiac surgery were randomised 1:1 to the following postoperative sedative regimens: a propofol infusion and dexmedetomidine (0.4 μg kg-1 h-1) or a propofol infusion and saline 0.9% (placebo group). The study drug was started at chest closure and continued for 10 h. The primary endpoint was in-hospital POD, assessed using the Confusion Assessment Method and chart review method. POD over the course of hospital stay occurred in 31/177 (18%) and 33/172 (19%) patients in the dexmedetomidine and placebo arm, respectively (P=0.687; odds ratio=0.89; 95% confidence interval, 0.52-1.54). The incidence of POD in the intensive care alone, or on the ward alone, was also not significantly different between the groups. Subjects in the dexmedetomidine group spent less median time in a delirious state (P=0.026). Median administered postoperative norepinephrine was significantly higher in the dexmedetomidine group (P<0.001). One patient in the dexmedetomidine group and 10 patients in the placebo group died in the hospital. Adding low-dose rate dexmedetomidine to a sedative regimen based on propofol did not result in a different risk of in-hospital delirium in older patients undergoing cardiac surgery. With a suggestion of both harm and benefit in secondary outcomes, supplementing postoperative propofol with dexmedetomidine cannot be recommended based on this study. NCT03388541.


Bifidobacterium-selective isolation and enumeration from chicken caeca by a modified oligosaccharide antibiotic-selective agar medium.
To determine the efficacy and selectivity of an acidified, antibiotic-selective, oligosaccharide-containing media for enumerating Bifidobacterium spp. from chicken caeca samples. Transoligosaccharide propionate agar medium (TOS) modified by addition of mupirocin (50 microg ml-1) and glacial acetic acid (1%, v/v), did not inhibit the growth of bifidobacteria compared with the control media yet inhibited the growth of Lactobacillus acidophilus, Lactobacillus gallinarum, Lactobacillus helveticus and Streptococcus gordonii. Addition of mupirocin (50 microg ml-1) and glacial acetic acid (1%, v/v) to TOS (TOS-AM50), is an effective selective medium for isolation and enumeration of Bifidobacterium spp. from chicken caeca samples. The development of an intestinal bifidobacteria-selective media contributes to the study of probiotics and prebiotics in poultry and potentially other species.


Clinical Application of in Vitro Embryo Production in the Horse.
The first reports of in vitro embryo production (IVEP) by conventional in vitro fertilization and intracytoplasmic sperm injection in horses date respectively from approximately 30 and 25 years ago. However, IVEP has only become established in clinical practice during the last decade. The initial slow uptake of IVEP was largely because the likelihood of success was too low to make it an economically viable means of breeding horses. During the last decade, the balance has shifted, primarily because of significant improvements in the efficiency of recovering immature oocytes from live donor mares (historically <25%; now >50%) and in the successful culture of zygotes to the blastocyst stage in vitro (historically <10%; now >20%). It has also been established that immature oocytes can be "held" at room temperature for at least 24 hours, allowing overnight transport to a laboratory with expertise in IVEP. Moreover, because in vitro-produced embryos can be cryopreserved with no appreciable reduction in viability, they can be shipped and stored until a suitable recipient mare is available for transfer. Most importantly, in an established equine ovum pick-up intracytoplasmic sperm injection (OPU-ICSI) program, blastocyst production rates now exceed 1 per procedure, and posttransfer foaling rates exceed 50%, such that overall efficiency betters that of either embryo flushing or oocyte transfer. Moreover, OPU-ICSI can be performed year round and allows embryo production from mares with severe acquired subfertility and extremely efficient use of scarce or expensive frozen semen. Cumulatively, these factors have stimulated rapid growth in demand for IVEP among sport horse breeders.


Nrf2 attenuates ferroptosis-mediated IIR-ALI by modulating TERT and SLC7A11.
Acute lung injury (ALI) carries a mortality rate of ~50% and is a hot topic in the world of critical illness research. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a critical modulator of intracellular oxidative homeostasis and serves as an antioxidant. The Nrf2-related anti-oxidative stress is strongly associated with ferroptosis suppression. Meanwhile, telomerase reverse transcriptase (TERT), the catalytic portion of the telomerase protein, is reported to travel to the mitochondria to alleviate ROS. In our study, we found that TERT was significantly reduced in lung tissue of Nrf2-/- mice in the model of intestinal ischemia/reperfusion-induced acute lung injury (IIR-ALI). In addition, MDA levels showed marked increase, whereas GSH and GPX4 levels fell drastically in ALI models. Moreover, typical-related structural changes were observed in the type II alveolar epithelial cells in the IIR model. We further employed the scanning transmission X-ray microscopy (STXM) to examine Fe levels and distribution within cells. Based on our observations, massive aggregates of Fe were found in the MLE-12 cells upon OGD/R (oxygen and glucose deprivation/reperfusion) induction. Additionally, Nrf2 silencing dramatically reduced TERT and SLC7A11 levels, and further exacerbated cellular injuries. In contrast, TERT-overexpressing cells exhibited marked elevation in SLC7A11 levels and thereby inhibited ferroptosis. Collectively, these data suggest that Nrf2 can negatively regulate ferroptosis via modulation of TERT and SLC7A11 levels. The conclusion from this study brings insight into new candidates that can be targeted in future IIR-ALI therapy.


Purification and properties of a constitutive beta-lactamase from Pseudomonas aeruginosa strain Dalgleish.
1. The beta-lactamase (penicillin amido-beta-lactamhydrolase EC 188.8.131.52) appeared to be periplasmic rather than truly intracellular, since it was released by freeze-thawing without gross morphological changes in the cell. 2. The partially purified enzyme had pI between 5.0 and 5.5, mol. wt 32 000 and a broad pH vs activity profile with a maximum at pH 8. 3. The cephalosporins tested were hydrolysed less rapidly than most of the penicillins, and the Km values for penicillins were lower than for cephalosporins. However cloxacillin was hydrolysed very slowly although it was strongly bound. The substrate-induced inactivation common to many beta-lactamases was particularly marked with cephaloridine and cloxacillinmthe cloxacillin-induced inactivation was shown to be reversible.


An increase in tumor size increases the risk of the development of postoperative cardiopulmonary events in patients undergoing partial nephrectomy.
The aim of this study was to examine the effect of kidney tumor size on the risk of CPE developing in the first postoperative month. Evaluation was made of 127 patients who underwent PN between January 2010 and November 2022. The patients were separated into two groups as Group 1 (n: 13) including patients who developed CPE within the first postoperative month and Group 2 (n: 114) of patients who did not develop CPE. The factors that could affect CPE and overall survival were analyzed with multivariate logistic and Cox regression analysis, respectively. The groups were determined to be similar in respect of age, gender and Charlson Comorbidity Index (p = 0.35, p = 0.68, p = 0.42, respectively). The values of mean tumor size (48.6 ± 12.9 vs. 29.2 ± 8.7 mm, p < 0.001), clinical T1b stage (61.5% vs. 9.6%, p < 0.001), median R.E.N.A.L. Nephrometry Score (9[3] vs.6 [1], p = 0.001) and mean warm ischaemia time (21.2 ± 3.5 vs. 15.9 ± 2.63 min, p < 0.001) were determined to be statistically significantly higher in Group 1 than in Group 2. In the ROC curve analysis performed to predict the development of CPE within the first postoperative month, 35.5 mm was determined to be the best cut-off point for tumor diameter (AUC = 0.88, p < 0.001). In the multivariate analysis, the presence of CPE for overall survival, and increased tumor size for the development of CPE were each determined to be independent risk factors (OR: 3.25, p = 0.03; OR: 1.4, p = 0.001, respectively). Tumor size serves as a significant marker for the development of CPE within the initial month following PN.


High incidence of angiosarcomas in brown-fat tissue and livers of mice fed sterigmatocystin.
BDF1 female mice fed 30 or 120 ppm sterigmatocystin in their diet for 55 or 51 wk, respectively, developed angiosarcomas of the liver and of the dorsal brown-fat tissue. No tumours were observed in five mice from each group, including the controls, killed after 43 wk, but angiosarcomas were observed in mice from the test groups that died or were killed after more than 43 wk of treatment with sterigmatocystin. Thirty-four out of 53 mice (64.2%) fed 30 ppm sterigmatocystin developed hepatic angiosarcomas. Six (11.3%) of those fed 30 ppm sterigmatocystin and 27 out of the 51 mice (52.9%) fed 120 ppm developed angiosarcomas in the dorsal brown-fat tissue. Benign vascular lesions in the liver and a few angiosarcomas of the ovary and lung were also observed in mice of both test groups. No vascular changes were observed in control mice except for peliosis-like lesions in the spleens of two mice. Increases in the incidences of lung and hepatocellular adenomas were also observed in sterigmatocystin-treated mice.


Tonsillar lymphangiomatous polyp in an adult dog.
A female spayed cocker spaniel dog, aged 8 years, developed a large, smooth pedunculated mass arising from the right palatine tonsil. Histology revealed that the mass was composed of many, variably dilated, thin-walled lymphatic channels filled with pale eosinophilic fluid lacking red blood cells and embedded in a dense, fibrovascular stroma. The dilated lymphatic channels were lined by a single layer of flattened, discontinuous endothelium with scattered intraluminal valves. Periodic acid-Schiff staining highlighted the discontinuous basement membrane and immunohistochemistry revealed strong cell membrane and cytoplasmic immunoreactivity for CD31 and von Willebrand factor, respectively. The clinical and histological findings were consistent with a tonsillar lymphangiomatous polyp - an uncommon, benign tumour of the tonsil in man that has previously been unrecognized in veterinary medicine.


Acquired hydrocephalus. III. A pathophysiological study correlated with neuropathological findings and clinical manifestations.
On the basis of the laws of Pascal and Laplace, it is shown that the ventricular dilatation in acquired hydrocephalus is due to a primary increase in the intraventricular pressure (IVP), and that a new steady state can be reached, whether the IVP is increased or normal. The pressure increase is due to a disproportion between the production and reabsorption of cerebrospinal fluid (CSF). As water and salts pass freely across the ependyma and the choroid plexus in hydrocephalus, the pressure increase is caused by an increased protein concentration in the ventricular CSF, leading to increased fluid contents according to the Gibbs-Donnan equilibrium. During the ventricular dilatation, the ependyma is destroyed, and the protein molecules penetrate into the subependymal part of the white matter. This results in a reduction in the colloid osmotic pressure of the ventricular CSF, and a new steady state can be reached, with a normal protein concentration in an increased volume. The attendant microscopic changes in the ventricular wall were demonstrated in a patient with acquired hydrocephalus, and the observations made were in conformity with the results of a number of animal experiments. The symptomatology of acquired hydrocephalus is in agreement with a primary affection of the axons running in the juxtaventricular part of the white matter.


Valorization of pistachio industrial waste: Simultaneous recovery of pectin and phenolics, and their application in low-phenylalanine cookies for phenylketonuria.
This study introduces a sustainable approach to simultaneously produce pectin and phenolic compounds from pistachio industrial waste and applies them in the formulation of low-phenylalanine cookies. The co-optimization process was performed using the microwave-assisted technique and a Box-Behnken design, considering four variables and two responses: pectin yield and total phenolic content (TPC). The co-optimized condition (microwave power of 700 W, irradiation time of 210 s, pH level of 1.02, and LSR of 20 mL/g) resulted in a pectin yield of 15.85 % and a TPC of 10.12 %. The pectin obtained under co-optimized condition was evaluated for its physicochemical, structural, and thermal properties and the phenolic extract for its antiradical activity. Characterization of the pectin sample revealed a high degree of esterification (44.21 %) and a galacturonic acid-rich composition (69.55 %). The average molecular weight of the pectin was determined to be 640.236 kDa. FTIR and 1H NMR spectroscopies confirmed the structure of pectin, with an amorphous nature and high thermal stability observed through XRD and DSC analysis. Additionally, the extract exhibited significant antiradical activity comparable to butylated hydroxyanisole and ascorbic acid. The isolated ingredients were used to formulate low-protein, low-phenylalanine cookies for phenylketonuria patients. The addition of 0.5 % pectin and 1 mL/g extract led to increased moisture content (from 9.05 to 12.89 %) and specific volume (from 7.28 to 9.90 mL/g), decreased hardness (from 19.44 to 10.39 N × 102), and improved antioxidant properties (from 5.15 % to 44.60 % inhibition) of the cookies. Importantly, there was no significant increase observed in the phenylalanine content of the samples with pectin and extract addition. Furthermore, sensory evaluation scores demonstrated significantly higher scores for taste, odor, texture, and overall acceptability in cookies enriched with 0.5 % pectin and 1 mL/g extract, with scores of 4.53, 3.93, 4.40, and 4.60, respectively.


Development and clinical application of a rapid, visually interpretable polymerase spiral reaction for tcdB gene of Clostridioides difficile in fecal cultures.
In the surveillance of outbreaks of Clostridioides difficile infection, the rapid detection and diagnosis of C. difficile remain a major challenge. Polymerase spiral reaction (PSR) is a nucleic acid amplification technique that uses mixed primers and the strand displacement activity of Bst DNA polymerase to achieve a pair of primers and a single enzyme in an isothermal environment. The primer design is simple, the reaction is efficient, and a color indicator can be used to visualize the result. In this study, we developed a rapid and visually interpretable PSR to detect C. difficile by analyzing artificially contaminated feces samples and clinical isolates from patient feces samples. We designed two pairs of primers for a PSR that specifically targeted the conserved tcdB gene of C. difficile. The amplification results were visualized with the chromogenic dye hydroxynaphthol blue. The entire process was accomplished in 50 min at 64°C, with high specificity. The limit of detection of C. difficile with PSR was 150 fg/μl genomic DNA or 2 × 10 CFU/ml in artificially contaminated feces samples. With this method, we analyzed four clinical isolates and also compared the PSR with an isolation-and-culture detection method, polymerase chain reaction, and the Sanger sequencing. The four clinical isolates were found positive for tcdB, which confirmed the high specificity of the primers. The positive rates of tcdB in toxigenic C. difficile detected with PSR, PCR, and Sanger sequencing were 100%. The proportions of toxin types in these clinical C. difficile strains were 50% tcdA+tcdB+CDT- and 50% tcdA+tcdB+CDT+. The assay described should extend our understanding of the incidence of C. difficile. This may allow the rapid diagnosis and screening of C. difficile-related disease outbreaks in the field.


Hepatitis B vaccination failure in children with diabetes mellitus? The debate continues.
The aim of our study was to evaluate the presence of specific antibodies against HBsAg in diabetic children (IDDM) previously vaccinated against hepatitis B virus. 110 diabetic children were retrospectively studied and 100 healthy controls were recruited. In all patients surface antigen, HBV core IgG, antibodies against HBV "e" antigen and quantitative HBV surface antibodies were detected. In 45 patients molecular typing of HLA alleles was performed. Metabolic control was evaluated as mean glycated hemoglobin (HbA1c) and all patients were compliant to insulin therapy. 46 of 110 diabetic children (41.8%) and 16 of 100 healthy controls (16%) were found to have not anti-HBs antibodies (p < 0.0001). The mean antibody titer was found significantly-lower (p < 0.0001) in IDDM children than healthy controls. No correlation was found between antibody titer, age, duration of disease and HbA1c. We did not find any difference of gender, age, years of onset of the disease and metabolic control, between diabetics with anti-HBs antibodies and those without. Our data confirm the reduced seroprotection rate for HBV vaccination in diabetics. However it remains poorly clarify the real clinical significance of this result. In our study no diabetic children showed markers of HBV infection.


Application of telehealth for comprehensive Creutzfeldt-Jakob disease surveillance in the United Kingdom.
Creutzfeldt-Jakob disease (CJD) is a fatal human prion disease. Surveillance systems operate globally with the goals of accurate in-life case ascertainment, appropriate public health interventions to minimise secondary transmission, and monitoring trends in disease epidemiology. The UK experienced the highest incidence of variant CJD (vCJD) in the world following widespread population exposure to bovine spongiform encephalopathy (BSE). 178 cases of vCJD have been identified in the UK by the National CJD Research & Surveillance Unit (NCJDRSU), including three cases of secondary transmission via blood transfusion. The NCJDRSU performs high-fidelity surveillance, assessing all cases of suspected CJD referred to the unit. COVID-19 has caused widespread disruption to healthcare and poses a threat to services. The NCJDRSU converted to telehealth-based surveillance in March 2020. We report the results of the application of telehealth for comprehensive CJD surveillance during the first four months of the pandemic. 59 cases were assessed for suspected CJD. In 52 cases the relatives were interviewed for an informant history, by video conference or telephone call. 34 patients underwent video examination; 1 case was examined in-person. MRI images were assessed in all cases and 46 underwent CSF testing. Feedback was obtained from interviewees and the NCJDRSU team on their experiences. 50 cases were diagnosed with sporadic CJD; 5 received an alternative diagnosis, and the remaining 4 remained unresolved, with further investigations underway. Telehealth significantly reduced time taken to assessment compared to in-person assessments in 2019. Telehealth is an effective way to provide comprehensive CJD surveillance at a national level.


Sialoglycoproteins and sialoglycolipids contribute to the negative surface charge of epimastigote and trypomastigote forms of Trypanosoma cruzi.
Epimastigote and trypomastigote forms of Trypanosoma cruzi have a net negative surface charge, as determined by direct measurement of the mean cellular electrophoretic mobility. Treatment of the parasites with neuraminidase reduces by 17 and 52% the mean electrophoretic mobility of epimastigote and bloodstream trypomastigote forms, respectively. Neuraminidase-treated cells recover their normal electrophoretic mobility if incubated for 2 h in the presence of fresh culture medium. The recovering process of epimastigotes is almost totally blocked by addition of inhibitors of either protein synthesis (puromycin) or N-glycosidically linked glycoprotein synthesis (tunicamycin). The recovering process of trypomastigotes is not totally inhibited by either puromycin or tunicamycin. Treatment of T. cruzi with trypsin reduces by 11 and 40% the mean electrophoretic mobility of epimastigote and bloodstream trypomastigote forms. Trypsin-treated cells recover their normal electrophoretic mobility if incubated for 4 h in fresh culture medium. The recovering process of trypomastigotes is partially inhibited by puromycin. The results obtained indicate that sialoglycoproteins and sialoglycolipids exist on the surface of T. cruzi, the latter being predominant on the surface of trypomastigotes.


Thymidylate synthetase from Diplococcus pneumoniae, properties and inhibition by folate analogs.
Thymidilate synthetase (methylenetetrahydrofolate:dUMP C-methyltransferase) in crude extract from Diplococcus pneumoniae exhibits a partial but variable requirement for Mg-2+ depending upon the buffer. Optimum Mg-2+ concentration is between 0.014 and 0.02 M. The optimum pH for activity in a variety of buffers occurred as a broad peak between 7.0 and 7.7. In Tris/acetate buffer, but not in potassium phosphate buffer, the pH optimum was different in the presence and absence of Mg-2+. Methylation of uridylate, cytidylate and deoxycytidylate could not be demonstrated over a pH range of 5.0-8.0. The enzyme exhibited an apparent Km for deoxyuridylate of 3.08 - 10-5 M and an apparent Km for L-(+)(minus)-5,10-methylene tetrahydrofolate of 2.66 - 10-4 M. During molecular-sieve chromatography and sucrose density-gradient centrifugation, the enzyme was detectable only as a single catalytically active form of Mr 34 000-38 000. 2,4-Diamino quinazoline antifolates were better competitive inhibitors (Ki = 3-8 -10-6 M) of thymidylate synthetase than 2,4-diamino pteridines (Ki = 3- 10-5 M). 2-Amino-4-hydroxy-quinazolines were the best inhibitors (Ki = 1.3-2.9 - 10-6 M). All of the 2,4-diamino quinazolines and pteridines inhibited dihydrofolate reductase from D. pneumoniae in a nearly stoichiometric fashion (Ki = less than 10-10 M). The 2-amino-4-hydroxy-quinazolines were poor inhibitors of this enzyme (Ki = 10=5 M).


A comparison of cake properties in traditional and turbulence promoter assisted microfiltration of particulate suspensions.
The use of turbulence promoter can effectively enhance the permeate flux in crossflow microfiltration (CFMF) of particulate suspensions. Flux enhancement which is generally attributed to the reduction in cake thickness, however, has still not been clearly understood. In this study, the effects of turbulence promoter on cake properties (thickness, porosity and particle size) were investigated during CFMF of calcium carbonate suspension. It indicates that turbulence promoter has important effects on cake properties that directly affect the cake resistance. The significant reduction in thickness and slight increase in porosity are positive to reduce the cake resistance. The remarkable decrease in particle size is the negative effect due to the increased specific resistance of a cake. As a whole, the overall cake resistance is still diminished by turbulence promoter and therefore permeate flux is improved. The theoretical calculation of cake resistance shows a good consistence with the experimental result. The cake properties in both cases (using a turbulence promoter or not) almost exhibit the similar trends under various operating conditions. Differently, the use of turbulence promoter can greatly alleviate the effects of transmembrane pressure or feed concentration on growth of cake layer and intensify the effects of inlet velocity on diminishing the particle deposition.


Cooperative activation of D1-like and D2-like dopamine receptors in the nucleus accumbens shell is required for the reinstatement of cocaine-seeking behavior in the rat.
Activation of D1-like (D1, D5) or D2-like (D1, D3, D4) dopamine receptors in the nucleus accumbens shell is sufficient to reinstate cocaine-seeking behavior in rats. The goal of these experiments was to assess whether cooperative activation of D1-like and D2-like dopamine receptors in the accumbens shell is required to promote cocaine reinstatement. Rats were initially trained to self-administer cocaine (0.25 mg, i.v.) using a fixed-ratio schedule of reinforcement for approximately 21 days. Animals subsequently underwent an extinction phase during which saline was substituted for cocaine. Once cocaine self-administration behavior was extinguished (defined as <15% of the total responses maintained during self-administration), dopamine receptor agonist-induced reinstatement of cocaine seeking was assessed. Administration of the selective D1/5 agonist R-(+)-6-chloro-7,8-dihydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrobromide (SKF-81297) (1.0 microg) or the D2/3 receptor agonist trans-(-)-(4aR)-4,4a,5,6,7,8,8a,9-octahydro-5-propyl-1H-pyrazolo[3,4-g]quinoline hydrochloride (quinpirole) (3.0 microg) directly into the nucleus accumbens shell promoted reinstatement of cocaine seeking. In order to determine if endogenous dopamine tone in the accumbens shell is required for dopamine receptor agonist-induced reinstatement of cocaine seeking, D1/5 or D2/3 dopamine receptor antagonists were administered into the nucleus accumbens shell prior to a selective dopamine receptor agonist. Microinfusion of the D2/3 dopamine receptor antagonist sulpiride ((S)-5-aminosulfonyl-N-[(1-ethyl-2-pyrrolidinyl)methyl]-2-methoxybenzamide) (1.0 microg) into the nucleus accumbens shell 10 minutes prior to SKF-81297 (1.0 microg) blocked the ability of this D1-like dopamine receptor agonist to reinstate cocaine seeking. Similarly, administration of the selective D1/5 dopamine receptor antagonist R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrochloride (SCH-23390) (1.0 microg) into the nucleus accumbens shell prior to quinpirole (3.0 microg) blocked reinstatement of drug-seeking behavior elicited by this D2/3 dopamine receptor agonist. Moreover, intra-accumbal shell co-administration of subthreshold doses of quinpirole (1.5 microg) and SKF-81297 (0.1 microg) promoted cocaine-seeking behavior. Collectively, these results indicate that cooperative activation of D1-like and D2-like dopamine receptors in the nucleus accumbens shell is necessary to reinstate cocaine seeking in rats.


Magnetic resonance imaging and clinical evaluation of patellar resurfacing with press-fit osteochondral autograft plugs.
Autologous osteochondral transplantation (AOT) has been successfully used in the femoral condyle and trochlea and is an attractive treatment option for full-thickness patellar cartilage lesions. Patients treated with AOT for the repair of symptomatic, isolated patellar cartilage lesions will demonstrate improvement in functional outcomes and postoperative magnetic resonance imaging appearance. Case series; Level of evidence, 4. Between 2002 and 2006, patients with focal patellar cartilage lesions treated with AOT were prospectively followed. The mean age at the time of surgery was 30 years. Clinical assessment was performed with the International Knee Documentation Committee (IKDC), activities of daily living of the Knee Outcome Survey (ADL), and Short Form-36 (SF-36) at baseline and most recent follow-up. Magnetic resonance imaging was used to evaluate the cartilage repair morphologic characteristics in 14 cases. Twenty-two patients met the study criteria with a mean follow-up of 28.7 months (range, 17.7-57.8 months). The mean patellar lesion size was 165.6 +/- 127.8 mm(2), and the mean size of the donor plug was 9.7 +/- 1.1 mm in diameter with 1.8 +/- 1.4 plugs/defect. The mean preoperative IKDC score was 47.2 +/- 14.0 and improved to 74.4 +/- 12.3 (P = .028). The mean preoperative ADL score was 60.1 +/- 16.9 and increased to 84.7 +/- 8.3 (P = .022). The mean SF-36 also demonstrated an improvement, from 64.0 +/- 14.8 at baseline to 79.4 +/- 15.4 (P = .059). Nine patients underwent concomitant distal realignment and demonstrated improvement between preoperative and postoperative outcomes scores, but these differences were not statistically significant. Magnetic resonance imaging appearance demonstrated that all plugs demonstrated good (67%-100%) cartilage fill, 64% with fissures < 2 mm at the articular cartilage interface, 71% with complete trabecular incorporation, and 71% with flush plug appearance. Patellar AOT is an effective treatment for focal patellar chondral lesions, with significant improvement in clinical follow-up. This study suggests that patients with patellar malalignment may represent a subset of patients who have a poor prognostic outlook compared with patients with normal alignment.


Estimation of the aqueous solubility of weak electrolytes.
Jain and Yalkowsky [Jain, N., Yalkowsky, S.H., 2001. Estimation of the aqueous solubility. I. Application to organic non-electrolytes. J. Pharm. Sci. 90, 234-252.] demonstrated that the general solubility equation (GSE) can be used to estimate the aqueous solubility of organic non-electrolytes. In this study the applicability of the GSE was extended to weak electrolytes. It is demonstrated that the GSE estimates the aqueous solubility of 949 compounds, including 367 weak electrolytes with an AAE of 0.58. It is also shown that the intrinsic solubilities of weak acids for which the pK(a)+log S(w)<or=0 and for weak bases for which pK(a)-log S(w)<or=14 are within a factor of 2 of the total solubilities.


A 3-D airway epithelial cell and macrophage co-culture system to study Rhodococcus equi infection.
We developed a 3-D equine bronchial epithelial cell (BEC) culture that fully differentiates into ciliary beating and mucus producing cells. Using this system, we evaluated how mucus affects the phagocytic activity of macrophages. Adult horse monocyte-derived macrophages were incubated with Rhodococcus equi for 4h either in the mucus layer of in vitro generated airway epithelium or on collagen coated membranes. Using light and electron microscopy, we noted that the number of macrophages with intracellular bacteria, and the number of intracellular bacteria per macrophage were lower in the presence of mucus. TNFα measurements revealed that the presence of BECs promoted TNFα production by R. equi-infected macrophages; a decrease in TLR-2 (involved in R. equi recognition) and an increase in EGF-R (involved in mucin production) mRNA expression were also noted. Interestingly, when foal macrophages were added to foal BECs, we made the opposite observation, i.e. many macrophages were loaded with R. equi. Our in vitro bronchial system shows great potential for the identification of mechanisms how BECs and mucus play a role in phagocyte activation and bacterial clearance. Further studies using this system will show whether the airway environment in the foal responds differently to R. equi infection.


[Effect of nifedipine on effective renal plasma flow, plasma renin activity and serum aldosterone, noradrenaline and adrenaline levels in healthy persons and in patients with primary moderate arterial hypertension].
Blood pressure, plasma renin activity, and serum aldosterone, adrenaline and noradrenaline were investigated in healthy individuals and patients with the primary moderate hypertension following a single oral dose of 10 mg nifedipine. It was found that the drug is hypotensive in both healthy individuals and hypertensive patients. It does not affect the effective plasma flow throughout the kidneys as well as serum aldosterone and adrenaline whereas serum noradrenaline and plasma renin activity are increased.


Gastric electrical stimulation for gastroparesis.
Recently, gastric electrical stimulation (GES) has been used to treat gastroparesis. This study analyzes a cohort of gastroparetic patients after GES. All patients undergoing GES from October 2003 to July 2007 were included. Pre- and postoperative assessments were performed for frequency and severity of gastrointestinal symptoms and gastric retention. The values were compared using a paired t-test for patients at 6 and 12 months. Statistical significance was defined as p < 0.05. Fifty gastroparetic patients were enrolled (20 diabetic, 25 idiopathic, 2 postsurgical, and 3 connective tissue disorder patients). All patients underwent laparoscopic implantation with GES (Medtronic, Inc). Median followup was 28 months (range 3 to 51 months). Thirty-five patients were available for followup at 6 months, and 30 patients were available at 12 months. The total symptom severity score (19.05+/-8.04) decreased significantly at 6 months (12.92+/-7.41, p < 0.001) and 12 months (14.05+/-8.28, p < 0.01). Similarly, total frequency score (20.39+/-8.08) decreased significantly at 6 months (15.01+/-7.37, p < 0.01) and 12 months (15.71+/-7.40, p < 0.05). At 12 months (n=27), gastric retention at 2 hours was decreased significantly from 66% +/- 21% to 50% +/- 22% (p < 0.04) and normalized in 11 of 27 patients. The severity of symptoms was reduced in all patients with normal gastric retention postoperatively. Finally, gastric retention at 4 hours was reduced by 14%, but the difference was not significant. Gastroparetic symptoms at 6 months were improved and sustained at 12 months after GES. Gastric emptying at 2 hours was reduced significantly after GES. Longterm followup of this cohort is required to confirm the short-term effects of GES.


Fish community responses to restoration of a eutrophic coastal bay.
Interest in coastal restoration measures is increasing, but information about subsequent ecosystem recovery processes is limited. In Björnöfjärden on the Baltic Sea coast, Stockholm archipelago, a pioneering case study to reduce coastal eutrophication led to improvements and initially halved phosphorus levels. Here, we evaluate the effects of the restoration on the local fish assemblage over one decade after the measures. The study gives a unique possibility to evaluate responses of coastal fish to nutrient variables and abatement in a controlled natural setting. Cyprinid abundance decreased and perch partially increased with decreasing turbidity levels, while mean trophic level increased over time in the restored area. Responses were overall weak, likely attributed to an attenuation of the eutrophication abatement effect over time. The results suggest that nutrient reduction gives slow responses in fish compared to alternative measures such as fishing closures.


Ecological trends in lichen photosynthesis.
Drawing on data for 42 fruticose and foliose lichen species, four aspects of photosynthetic response have been ex&mined in relation to latitude and climatic variables: 1) the maximal rate of net photosynthesis (Pmax), 2) the tissue temperature at which Pmax occurs (Topt), 3) the photon flux density at which Pmax occurs (PhARsat), and 4) the tissue water status at which Pmax occurs. Tissue water status was measured as either relative water content (RWCopt) or percentage dry weight (DWopt). The Topt decreased significantly with increasing latitude; Topt also increased significantly with increasing July solar radiation, July air temperature, daily hours of bright sunshine, daily solar radiation, annual precipitation, and annual evaporation. Both RWCopt and DWopt increased significantly with annual precipitation. The Topt and RWCopt for a lichen could both be more accurately predicted from multiple regressions on macroclimatic variables describing the lichen's habitat. Both the PhARsat and the Pmax, despite their lack of simple univariate relationships to either latitude or climatic variables, could also be predicted by multiple regression. The limitations on the generality and precision of these ecological trends in lichen photosynthesis are discussed in relation to microclimatic considerations, morphological adaptations, and poikilohydric metabolism.


The molecular basis of the Rhesus antigen Ew.
The Rhesus antigen Ew (ISBT designation 004 011) was first described in 1955. It is defined by a specific antibody, but its molecular genetic basis has not yet been resolved. Two individuals serologically characterized to express the rare Rhesus antigen Ew were analyzed by sequencing of all 10 exons of the RHCE gene. A nucleotide exchange at position 500 (T500A) resulting in a Met167Lys amino acid substitution was found in both individuals. Moreover, we show that an individual carrying the Ew antigen is capable to produce an alloantibody against the wild-type E antigen. The single-point mutation T500A in exon 4 of the RHCE gene is a molecular basis of the rare Rhesus antigen Ew.


CT analysis of anatomical distribution of melorheostosis challenges the sclerotome hypothesis.
Melorheostosis (MEL) is a rare disease of high bone mass with patchy skeletal distribution affecting the long bones. We recently reported somatic mosaic mutations in MAP2K1 in 8 of 15 patients with the disease. The unique anatomic distribution of melorheostosis is of great interest. The disease remains limited to medial or lateral side of the extremity with proximo-distal progression. This pattern of distribution has historically been attributed to sclerotomes (area of bone which is innervated by a single spinal nerve level). In a further analysis of our study on MEL, 30 recruited patients underwent whole body CT scans to characterize the anatomic distribution of the disease. Two radiologists independently reviewed these scans and compared it to the proposed map of sclerotomes. We found that the disease distribution conformed to the distribution of a single sclerotome in only 5 patients (17%). In another 12 patients, the lesions spanned parts of contiguous sclerotomes but did not involve the entire extent of the sclerotomes. Our findings raise concerns about the sclerotomal hypothesis being the definitive explanation for the pattern of anatomic distribution in MEL. We believe that the disease distribution can be explained by clonal proliferation of a mutated skeletal progenitor cell along the limb axis. Studies in mice models on clonal proliferation in limb buds mimic the patterns seen in melorheostosis. We also support this hypothesis by the dorso-ventral confinement of melorheostotic lesion in a patient with low allele frequency of MAP2K1-positive osteoblasts and low skeletal burden of the disease. This suggests that the mutation occurred after the formation of dorso-ventral plane. Further studies on limb development are needed to better understand the etiology, pathophysiology and pattern of disease distribution in all patients with MEL.


An isoform of Nedd4-2 is critically involved in the renal adaptation to high salt intake in mice.
Epithelial sodium channels (ENaCs) play critical roles in the maintenance of fluid and electrolyte homeostasis, and their genetic abnormalities cause one type of hereditary salt-sensitive hypertension, Liddle syndrome. As we reported previously, both human and rodent Nedd4L/Nedd4-2 showed molecular diversity, with and without a C2 domain in their N-terminal. Nedd4L/Nedd4-2 isoforms with a C2 domain are hypothesized to be related closely to ubiquitination of ENaCs. We generated Nedd4-2 C2 domain knockout mice. We demonstrate here that loss of Nedd4-2 C2 isoform causes salt-sensitive hypertension under conditions of a high dietary salt intake in vivo. The knockout mice had reduced urinary sodium excretion, osmotic pressure and increased water intake and urine volume with marked dilatation of cortical tubules while receiving a high salt diet. To the contrary, there was no difference in metabolic data between wild-type and knockout mice receiving a normal control diet. In the absence of Nedd4-2 C2 domain, a high salt intake accelerated ENaC expression. Coimmunoprecipitation studies revealed suppressed ubiquitination for ENaC with a high salt intake. Taken together, our findings demonstrate that during a high oral salt intake the Nedd4-2 C2 protein plays a pivotal role in maintaining adaptive salt handling in the kidney.


Tc-99m diethylenetriamine pentaacetic acid (DTPA) renal function reserve estimation: is it a reliable predictive tool for assessment of preclinical renal involvement in scleroderma patients?
Prognosis of systemic sclerosis (SSc) depends on internal organ involvement. We assessed the value of renal function reserve (RFR) for the detection of preclinical nephropathy in scleroderma. Thirty SSc patients with normal serum creatinine and 30 healthy controls were included. Medsger disease severity score, glomerular filtration rate (GFR), and microalbuminuria were measured. Tc-99m DTPA was utilized for GFR measurement at baseline and after oral protein overload (stimulated GFR). RFR was calculated as the percentile increase of stimulated GFR. SSc patients had lower means of baseline GFR (P=0.001), stimulated GFR (P=0.004), RFR (P=0.046), and higher microalbuminuria (P=0.009) than controls. According to baseline GFR, SSc patients showed three categories-normal baseline GFR (n=12), hyperfiltration GFR (n=3), and reduced baseline GFR (n=15). In the former category, RFR was normal in 6/12 patients and abnormal in the remainders (50%). Hyperfiltration patients and those with reduced baseline GFR showed abnormal RFR. A statistically significant negative association was found between microalbuminuria versus stimulated GFR and RFR (r= -0.5, P=0.007 and r= -0.45, P=0.013, respectively). The majority of SSc patients with abnormal RFR had disease duration of ≥48 months (60% vs. 20%, P=0.008). All SSc patients with pulmonary hypertension had abnormal RFR, while reduced baseline GFR was noted in only 60%. A significant negative correlation was found between reduced baseline GFR and cumulative dose of corticosteroids in SSc patients (r= -0.4, P=0.022). RFR estimation could be a useful predictive marker for preclinical renal involvement in SSc patients so that early prophylactic measures and therapy modifications could be considered.


Who Needs Level III Lymph Node Dissection in Carcinoma Breast-Study from a Tertiary Care Center.
In Indian females, breast cancer is the most common cancer with a late stage of presentation leading to one-third of patients undergoing modified radical mastectomy (MRM). Our study is undertaken to find out predictors of level III axillary lymph node metastasis in breast cancer and who needs complete axillary lymph node dissection (ALND). Retrospective study of 146 patients who undergone MRM or breast-conserving surgery (BCS) with complete ALND at Kidwai Memorial Institute of Oncology was done, and data was analyzed to find out the frequency of level III lymph nodes and the demographic relation and its relation to positive lymph nodes in level I + II. Positive metastatic level III lymph node was found in 6% of patients, with the median age of the patient in our study with level III positivity was 48.5 years with 63% pathological stage II with 88% perinodal spread (PNS)- and lymphovascular invasion (LVI)-positive. Involvement of level III lymph node was associated with gross disease in level I + II lymph node having more than four lymph node-positive and with pT3 stage or more which has higher chances of level III lymph node involvement. Level III lymph node involvement, though rare in early-stage breast cancer, is associated with larger clinical and pathological sizes (T3 or more), more than 4 lymph node-positive in level I + II and with PNS and LVI. Hence, based on these results, we recommend that for inpatient with more than 5-cm tumor size and those with the gross disease in axilla, complete ALND is recommended.


Basics of compounding: considerations for implementing United States pharmacopeia chapter 797 pharmaceutical compounding-sterile preparations, part 20: patient or caregiver training, patient monitoring and adverse events reporting, and quality-assurance program.
This article is intended to address the activities of the caregiver and encourage the need to address adverse events or medical errors. The advantage of disclosing adverse events or medication errors is to avoid further potential errors and determine/correct the cause of the errors. There should be awareness between patients and caregivers on the proper use and administration of intended compounded sterile preparations. Patients/patient's relatives should ask questions about their medications and be knowledgeable or cognizant on the appearance of the preparation or specified dosage of the medication. Caregivers should be open and explicit about the medications being administered or prescribed by the doctor and the rationale for such medications. This module will address these sections in a practical manner and provide practice clarification to the standards.


GW9662, a peroxisome proliferator-activated receptor gamma antagonist, attenuates the development of non-alcoholic fatty liver disease.
Insulin resistance is among the key risk factors for the development of non-alcoholic fatty liver disease (NAFLD). Recently, it has been reported that GW9662, shown to be a potent peroxisome proliferator-activated receptor gamma (PPARγ) antagonist, may improve insulin sensitivity in settings of type 2 diabetes. Here, we determined the effects of GW9662 on the development of NAFLD and molecular mechanisms involved. Female C57BL/6J mice were pair-fed either a liquid control diet (C) or a fat-, fructose- and cholesterol-rich diet (FFC) for 8 weeks while either being treated with GW9662 (1 mg/kg body weight; C+GW9662 and FFC+GW9662) or vehicle (C and FFC) i.p. three times weekly. Indices of liver damage and inflammation, parameters of glucose metabolism and portal endotoxin levels were determined. Lipopolysaccharide (LPS)-challenged J774A.1 cells were treated with 10 μM GW9662. Despite similar caloric intake the development of NAFLD and insulin resistance were significantly attenuated in FFC+GW9662-treated mice when compared to FFC-fed animals. Bacterial endotoxin levels in portal plasma were almost similarly increased in both FFC-fed groups while expressions of toll-like receptor 4 (Tlr4), myeloid differentiation primary response 88 (Myd88) and interleukin 1 beta (Il1b) as well as nitrite (NO2-) concentration in liver were significantly higher in FFC-fed mice than in FFC+GW9662-treated animals. In J774A.1 cells, treatment with GW9662 significantly attenuated LPS-induced expression of Il1b, interleukin 6 (Il6) and inducible nitric oxide synthase (iNos) as well as NO2- formation. In summary, our data suggest that the PPARγ antagonist GW9662 attenuates the development of a diet-induced NAFLD and that this is associated with a protection against the activation of the TLR4 signaling cascade.


Detection of Shiga toxin-producing and other diarrheagenic Escherichia coli by the BioFire FilmArray® Gastrointestinal Panel in human fecal samples.
The purpose of this investigation was the evaluation of the performance of the BioFire FilmArray® Gastrointestinal (FA-GI) Panel, a multiplexed molecular stool screening assay, for the detection of diarrheagenic Escherichia coli (DEC), with emphasis on Shiga toxin-producing E. coli (STEC). A dilution series of 12 STEC reference strains was tested with the FA-GI Panel to assess the analytical sensitivity. A total of 389 patient samples were analyzed with the FA-GI Panel and confirmation of the detected DEC was attempted with an in-house culture-based polymerase chain reaction (PCR) method. All Shiga toxin genes, except the one encoding Stx2f, were detected in bacterial dilutions ranging from 10(4) to 10(2) colony-forming units (CFU)/ml. eae + stx2f + STEC was misclassified as enteropathogenic E. coli (EPEC). Different sensitivities for various gene targets present in one isolate led to differing identifications depending on the concentration. Using the in-house method as a reference, the FA-GI Panel had a sensitivity of 90.6 % [confidence interval (CI) 75.0 %-98.0 %] and a specificity of 97.2 % (CI 94.9 %-98.6 %) for STEC detection in feces. At least one DEC was reported in 35.5 % (171/389) of the patient specimens, with EPEC being the most prevalent (n = 71). Only 59.7 % of the detected DEC could be confirmed, presumably because the comparator method was not applied directly on feces. The FA-GI Panel could not detect the stx2f subtype, misclassified certain pathogens, and the high detection rate of EPEC needs further investigation. Nevertheless, we believe that this sensitive and convenient system will prove to be an invaluable tool for the rapid diagnosis of most DEC infections, but culturing of the detected microorganisms should always be attempted.


Anti-biofouling Ti3C2TX MXene-holey graphene modified electrode for dopamine sensing in complex biological fluids.
With the development of new methods for the medical diagnosis, electrochemical sensors have attracted increasing attention. However, biofouling on the surface of the sensor significantly decreases sensor performance, thereby limiting the application of electrochemical sensors in complex biological fluids. Given the urgent need for anti-biofouling electrodes, a sensor based on a glass carbon electrode (GCE) modified with Ti3C2TX MXene and electrochemically reduced holey graphene (ERHG) was fabricated and demonstrated to have excellent electrochemical performance and anti-biofouling properties. ERHG provides abundant surface-active sites and imparts stability by hindering the agglomeration and oxidation of MXene. Furthermore, the excellent conductivity and hydrophilicity of MXene result in a high electron transfer rate and strong hydrophilicity. The MXene-ERHG/GCE sensor can detect dopamine with a wide linear range (0.2-125 μM) and a low detection limit of 0.044 μM in phosphate-buffer saline solution. Importantly, the hydrophilicity of MXene-ERHG reduces non-specific protein adsorption on the electrode surface, providing resistance to biofouling. After immersion in bovine serum albumin for 30 min, MXene-ERHG/GCE retained 85.90% of its initial peak current value, much higher than that of ERHG/GCE (17.75%). The MXene-ERHG/GCE sensor also showed good sensitivity for dopamine detection in serum and artificial cerebrospinal fluid (aCSF) containing bovine serum albumin. Moreover, MXene-ERHG/GCE exhibited excellent reproducibility and long-term stability in aCSF. The results demonstrate that MXene-ERHG/GCE has excellent anti-biofouling performance, and shows potential as an electrode material for application in biosensing.


Early safety of tenofovir alafenamide in patients with a history of tubulopathy on tenofovir disoproxil fumarate: a randomized controlled clinical trial.
The aim of the study was to assess the effect of tenofovir alafenamide (TAF) on kidney and bone biomarkers in patients who developed proximal renal tubulopathy (PRT) while receiving tenofovir disoproxil fumarate (TDF). Individuals with a history of TDF-associated PRT and currently suppressed HIV infection on a tenofovir-sparing regimen were randomized 1:1 to continue current antiretroviral therapy or initiate emtricitabine (F)/TAF with discontinuation of nucleoside reverse transcriptase inhibitors (NRTIs) as appropriate. Renal and bone biomarkers were analysed at baseline, week 4 and week 12. The primary outcome was the mean difference between study arms in urine retinol-binding protein:creatinine ratio (RBPCR) change from baseline to week 12. Data were analysed using linear regression, with robust standard errors (primary outcome), and repeated measures mixed effects models (secondary outcomes). The trial was registered under European Union Drug Regulating Authorities Clinical Trials Database 2016-003345-29. We randomized 31 individuals [mean age 52.4 (standard deviation 0.3) years; 97% male; 90% white); all completed the study. At 12 weeks, there was no difference in change in RBPCR (β 19.6; 95% confidence interval -35.3, 74.5; P = 0.47), and no difference in change in estimated glomerular filtration rate (eGFR) (based on creatinine or cystatin C), albuminuria, proteinuria, renal phosphate or urea handling, (fasting) urine osmolality, parathyroid hormone and bone turnover markers in the control versus the F/TAF exposed groups. No cases of PRT were observed. In people with a history of proximal renal tubulopathy while on TDF, 12-week exposure to TAF did not adversely affect renal tubular function. These data support continued evaluation of the long-term safety of TAF in this group of patients.


Observational study in cardiac rehabilitation groups phase III: a comparison of perceived and measured training intensity during a moderate-intensity workout.
During the last 25 years, the absolute number of cases of cardiovascular disease has increased in Europe. The goal in cardiac rehabilitation (CR) groups phase III is to motivate participants to demonstrate exercises independently of the group and self-reliant in long-term. In this context, it is of great importance to teach participants the appropriate training intensity. Adjusting the intensity of the training in CR groups is essential in order to avoid the dangers of overexertion and undertraining. This study aimed to examine whether participants in CR groups could correctly assess their exertion and thus achieve their predefined training intensity. Observational study. The setting of the study was an outpatient rehabilitation center. Forty-one people from CR groups phase III, with cardiovascular diseases, were included in the study. The training focused on moderate-intensity activity. The Borg Scale was used to evaluate how participants rated their exertion. The heart rate was recorded with the Polar M600 during the entire training period to compare it with the estimated exertion. Sixty-five percent (65%) of the participants was trained within their prescribed training intensity. However, they rated the perceived exertion significantly higher than the measured HR (P<0.001). The difference between target training HR and measured HR was also significant (P=0.008). There were significant differences between the groups of moderate-intensity endurance training and moderate-intensity strength endurance training (P=0.002). The participants in the CR groups could assess their own exertion moderately well. On the one hand, there were difficulties in accurately estimating the perceived exertion compared to the actual exertion. On the other hand, overloading and underloading were hardly detected. The perception of exertion must be improved by knowing one's own training heart rate and intensity. Therefore, wearables such as smartwatches could be useful monitoring aids for CR groups to check if the predefined training intensity will be reached.