Miniaturized 3D printed electrochemical platform with optimized Fibrous carbon electrode for non-interfering hypochlorite sensing.
3D printing technology based electrochemical device can provide ease of fabrication, cost effectiveness, rapid detection and lower limit of detection. Herein, a novel, customized, portable and inexpensive 3D printed electrochemical device, has been presented. Fibrous carbon Toray paper, deposited with gold nanoparticles through electrodeposition, used as a working electrode which Further device was tested with 1 mM sodium hypochlorite using cyclic voltammetry (CV) and square wave voltammetry (SWV) in 0.1 M PBS. Hypochlorite has a pivotal role in supporting the growing chemical and paper industries and finds diverse uses in several clinical applications. It is primarily used for disinfecting food, water and surfaces. The scan rate study was carried out from 20 mVs-1 to 250 mVs-1 using cyclic voltammetry technique. The diffusion coefficient obtained from scan rate effect was 1.39 × 10-6 cm2s-1. The concentration range was evaluated with SWV technique, in a linear range of 0.6 μM-40 μM, with a detection limit of 0.7 μM. The device was further analyzed to ensure non-interference from co-existing chemicals like sodium chloride, potassium nitrate, sodium carbonate, sodium nitrite. Real sample analysis was done with sea, artificial sea and tap water with impressive recovery values. In summary, the developed working electrode can be customized and modified based on testing analyte; thus, the proposed device can be used for various other biochemical analytes.


Gefitinib (Iressa, ZD 1839) for non-small cell lung cancer (NSCLC): recents results and further strategies.
Phase I and II clinical studies demonstrated that gefinitib single agent therapy is well tolerated and provides clinically significant antitumor activity in patients with advanced NSCLC who have previously received prior treatment with cytotoxic chemotherapy. Large-scale studies to assess the clinical benefit of gefitinib versus placebo when combined with cisplatin/gemcitabine and carboplatin/paclitaxel as first-line treatment, however, did not prove that the three drug combinations be able to improve survival and compared to "standard"-doubled chemotherapy. Therefore it can be summarized that gefinitib may provide evaluable addition to the therapeutic options available for the treatment of advanced NSCLC and may also have potential for use in early NSCLC.


The Impact of Mental Illness on Postoperative Outcomes Among Medicare Beneficiaries: A Missed Opportunity to Help Surgical Patients?
The aim of this study was to define the prevalence of preexisting mental illness, as well as characterize the impact of a preexisting mental illness diagnosis on postoperative outcomes. Preoperative surgical evaluation and risk stratification have traditionally centered on optimizing physical health. The influence of mental health on postoperative surgical outcomes has not been investigated. Medicare beneficiaries who underwent elective colectomy, coronary artery bypass grafting, abdominal aortic aneurysm repair, abdominal aortic aneurysm repair, total hip arthroplasty, total knee arthroplasty, and lung resection were identified. Patients were classified as having mental illness using International Classification of Diseases, 9th and 10th Revisions Procedures codes (ICD9/10CM) codes for anxiety, depression, bipolar disorder, schizophrenia, or other psychotic disorder. Among 1,889,032 Medicare beneficiaries who met inclusion criteria 560,744 (n = 29.7%) individuals had an antecedent diagnosis of mental illness. The majority had anxiety and/or depression (91.8%), whereas a smaller subset (8.2%) had a severe mental illness diagnosis. Patients who did and did not have mental illness were comparable relative to age and comorbidities. Patients with preoperative mental illness had a higher chance of surgical complications [anxiety/depression odds ratio (OR) 1.44, 95% confidence interval (CI) 1.42-1.46, severe mental illness OR 1.71, 95%CI 1.66-1.77] and an extended length of stay (anxiety/depression OR 1.45, 95% CI 1.44-1.46, severe mental illness OR 2.34, 95% CI 2.28-2.39). History of anxiety/depression (OR 1.87, 95% CI 1.85-1.90) or severe mental illness (OR 2.86, 95% CI 2.77-2.94) was also associated with higher odds of 30-day readmission. Additionally, individuals with mental illness had a higher prevalence of suicidal ideation within the first year after surgery (no mental illness: 1.88 per 100,000 individuals; anxiety/depression: 51.3 per 100,000 individuals; severe mental illness: 238.9 per 100,000 individuals). Three in 10 Medicare beneficiaries had a preexisting mental illness diagnosis, which was strongly associated with worse postoperative outcomes, as well as suicide risk. Surgeons need to optimize mental health assessment and services in the preoperative setting to improve outcomes for this vulnerable population.


Clinical and epidemiologic characteristics of norovirus GII.4 Sydney during winter 2012-13 in Beijing, China following its global emergence.
Limited information is available on the molecular epidemiology of GII.4 Sydney-associated diarrhea in China in the winter of 2012-13 during the global epidemic associated with the emergence of GII.4 Sydney. Fecal specimens collected from 171 diarrhea outpatients (one from each) between late October 2012 and the middle of March 2013 were examined for NoV by reverse transcription-polymerase chain reaction and sequences corresponding to both the NoV partial polymerase and partial capsid regions were analyzed phylogenetically. Clinical characteristics of GII.4 Sydney cases versus other NoV-positive cases detected in a previous study were compared statistically. Twenty-six (15.2%, 26/171) outpatients with diarrhea were infected with NoV. Twenty-two of the 26 (84.6%) identified NoV strains clustered into GII.4 Sydney. There was a significant difference in symptoms of fever (χ(2), P<0.05 ), abdominal pain (χ(2), P<0.05 ) and diarrhea frequency (Mann-Whitney U test, P<0.05) between the GII.4 Sydney case group and other NoV-positive case group. The new NoV variant, GII.4 Sydney, has been circulating in Beijing, China and became the predominant strain in the winter of 2012-13. GII.4 Sydney causes severe fever, abdominal pain and higher diarrhea frequency clinically compared to other NoV infections.


Epigenome-wide DNA methylation analysis of whole blood cells derived from patients with GAD and OCD in the Chinese Han population.
Generalized anxiety disorder (GAD) and obsessive-compulsive disorder (OCD) had high comorbidity and affected more than 44 million people around the world leading to a huge burden on health and economy. Here, we conducted an epigenome-wide DNA methylation study employing 93 patients with GAD, 65 patients with OCD, and 302 health controls, to explore epigenetic alterations associated with the onset and differences of GAD and OCD. We identified multiple differentially methylated positions (DMPs) and regions (DMRs): three DMP genes included RIOK3 (cg21515243, p = 8.00 × 10-10), DNASE2 (cg09379601, p = 1.10 × 10-9), and PSMB4 (cg01334186, p = 3.70 × 10-7) and two DMR genes USP6NL (p = 4.50 × 10-4) and CPLX1 (p = 6.95 × 10-4) were associated with the onset of GAD and OCD; three DMPs genes included LDLRAP1 (cg21400344, p = 4.40 × 10-12), ACIN1 (cg23712970, p = 2.98×10-11), and SCRT1 (cg25472897, p = 5.60 × 10-11) and three DMR genes WDR19 (p = 3.39 × 10-3), SYCP1 (p = 6.41 × 10-3), and FAM172A (p = 5.74 × 10-3) were associated with the differences between GAD and OCD. Investigation of epigenetic age and chronological age revealed a different epigenetic development trajectory of GAD and OCD. Conclusively, our findings which yielded robust models may aid in distinguishing patients from healthy controls (AUC = 0.90-0.99) or classifying patients with GAD and OCD (AUC = 0.89-0.99), and may power the precision medicine for them.


Near-unity photoluminescence quantum yield in MoS₂.
Two-dimensional (2D) transition metal dichalcogenides have emerged as a promising material system for optoelectronic applications, but their primary figure of merit, the room-temperature photoluminescence quantum yield (QY), is extremely low. The prototypical 2D material molybdenum disulfide (MoS2) is reported to have a maximum QY of 0.6%, which indicates a considerable defect density. Here we report on an air-stable, solution-based chemical treatment by an organic superacid, which uniformly enhances the photoluminescence and minority carrier lifetime of MoS2 monolayers by more than two orders of magnitude. The treatment eliminates defect-mediated nonradiative recombination, thus resulting in a final QY of more than 95%, with a longest-observed lifetime of 10.8 ± 0.6 nanoseconds. Our ability to obtain optoelectronic monolayers with near-perfect properties opens the door for the development of highly efficient light-emitting diodes, lasers, and solar cells based on 2D materials.


A new BF F variant by polyacrylamide gel isoelectric focusing.
Polyacrylamide gel isoelectric focusing followed by electroblotting with enzyme immunoassay was done for the investigation of allotypes of properdin factor B (BF) in serum from 326 Japanese subjects. A new BF F variant tentatively designated BF*Fb1 (b = basic) was detected, the isoelectric point of each band of homozygous BF Fb1 being higher than of BF FF. Family data were in accordance with transmission by mendelian inheritance. The allele frequencies calculated from 326 Japanese subjects were 0.7945, 0.1825, 0.0215, and 0.0015 for BF*S, BF*F, BF*Fb1, and BF*F075, respectively, with that of variant BF*Fb1 being a polymorphic frequency. The distribution of phenotypes fitted the Hardy-Weinberg equilibrium.


Distribution of CFTR mutations in Saguenay- Lac-Saint-Jean: proposal of a panel of mutations for population screening.
Saguenay-Lac-Saint-Jean is a region located in the northeastern part of the Province of Quebec, Canada, and is characterized by a founder effect. In this region, it has been documented that the incidence of cystic fibrosis reached 1/902 live births between 1975 and 1988, three times higher than the average incidence of 1/2500 live births reported in other Caucasian populations. This corresponds to a carrier rate of 1/15. Using genotyping data from the Canadian Consortium for Cystic Fibrosis Genetic Studies, this article describes the cystic fibrosis transmembrane conductance regulator profile of the cystic fibrosis population living in the Saguenay-Lac-Saint-Jean region and compares it with cystic fibrosis populations living in three other regions of the Province of Quebec. Significant differences in allelic frequencies of common mutations (as DeltaF508, 621 + 1G>T and A455E), and in percentage of covered allele with three or six mutations, were found in Saguenay-Lac-Saint-Jean compared to other regions. Based on this result, two mutation panels exceeding 90% sensitivity threshold are now proposed for cystic fibrosis carrier screening in this region. The implementation of the proposed carrier screening program could diminish the incidence of this disease in this region and allow future parents to make informed decisions about family planning.


Improving primary mental health care: impact of a nationwide programme.
To determine the impact of a nationwide programme on primary mental health care practices. Cohort study involving two surveys conducted in 2001 and 2003. Random sample of 2757 general practitioners. In 2001, a nationwide programme was initiated to improve primary mental health care. The programme used a participatory approach with regional needs assessment and regional selection of the interventions. Perceived need, availability and participation in the interventions; self-reported mental health performance (self-reported diagnosis and management of mental health disorders); perceived barriers to optimal care. Baseline and follow-up questionnaires were returned by 1336 and 1358 general practitioners, respectively. Interventions did not entirely match general practitioners' needs. Mental health performance did not change consistently while perceived barriers to optimal care decreased considerably. Actual participation in the interventions was not associated with improved mental health performance or decreased perception of barriers. The application of the nationwide programme had no considerable impact on the professional practices of general practitioners with regard to mental health care. This finding challenges the assumption that a bottom-up approach to quality improvement is effective. Extensive coordination at a national level and the provision of specific supportive products and services appears to be necessary to achieve improvement.


Radiosulphate: its metabolism and use in measurement of extracellular fluid volume in calves.
Radiosulphate injected intravenously into calves under 14 days of age equilibrated, by 30 min, with a mean fluid volume of 311 ml/kg body weight. During the equilibration period 3-7 per cent of the injected dose was excreted in the urine. The volume of distribution of radiosulphate increased for at least 4-5 h after equilibration suggesting that it penetrated beyond the readily miscible water of the extracellular fluid.


Systemic Immune-Inflammation Index (SII) Can Be an Early Indicator for Predicting the Severity of Acute Pancreatitis: A Retrospective Study.
Systemic immune-inflammation index (SII) is a new systemic inflammatory prognostic indicator associated with outcomes in patients with different tumors. Studies have shown an association between SII and many chronic/acute inflammatory diseases. This study aimed at exploring whether SII can be used as an effective parameter for predicting the severity of acute pancreatitis (AP). A total of 101 acute pancreatitis patients were enrolled in this study (mild acute pancreatitis (MAP): n = 73 and severe acute pancreatitis (SAP): n = 28). Patient demographics and SII were analyzed using the chi-square test, Student's t-test, and Mann-Whitney U-test. A receiver operating characteristic curve was generated to test the potential of using neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), and SII to predict AP's severity. Logistic regression analysis was performed to determine major risk factors. Patients with SII value ≥2207.53 had a higher probability of having SAP (sensitivity = 92.9%, specificity = 87.7%, and AUC = 0.920), and SII was a significantly better predictive value than PLR and NLR. Logistic regression analysis results showed SII could differentiate MAP from SAP as a major risk factor. This study has shown that SII is a potential indicator for predicting the severity of acute pancreatitis. The findings suggested that SII is more sensitive and specific than NLR and PLR in predicting the severity of acute pancreatitis.


Neurohypophyseal hormone-responsive renal adenylate cyclase. IV. A random-hit matrix model for coupline in a hormone-sensitive adenylate cyclase system.
A "random-hit" matrix model is proposed to account for the dynamic and steady state relationship between occupation of bovine renal medullary membrane receptors by [Lys8]vasopressin (LVP) and neurohypophyseal hormones (NHH) and the associated activation of membrane-bound adenylate cyclase. The model was developed by systematic introduction of specific rules concerning receptor coupling into a general structural model which consists of two square matrices of identical size, one composed of homogeneous R ("receptor") units, the second of homogeneous C ("cyclase") units. R units are either occupied (RO) or unoccupied (RU); C units are either active (CA) or inactive (CI). Hormone molecules are envisioned to "collide" with R units randomly; collision with RU leads to "binding", and occupation is maintained for a characteristic mean occupancy time, TO. In this structure, each R unit has an "interaction field" which consists of the "twin" unit in the "C" matrix, and the 4 nearest neighbor C units surrounding the twin. Occupation of an R unit leads to activation of all CI units in the interaction field of that R; CA units in the interaction field are refractory. Thus binding at a given R may "recruit" a variable number of inactive neighboring C units (5, 4, 3, 2, 1, or 0). The model requires that there be individual coupling delays between the moment of binding at a given R and subsequent activation of CI units (mean coupling delay (Td) approximately 10% To). Activation of C units persists as long as the "parent" R is occupied and is maintained for an additional short time interval (Tp) after RO reverts to RU, corresponding to hormone dissociation from receptor. The model accounts for the following previously demonstrated relations between LVP occupation of receptors and adenylate cyclase activation in bovine renal medullary membranes: 1) the shape of the nonlinear steady state relation between normalized (percentage maximal) receptor occupation (O) and cyclase activation (A), uniformly observed in different membrane preparations: 2) variable hormone concentration-dependent trajectories of approach to the final steady state A:O value (A:Oss) which may be either monophasic or biphasic; 3) the loss of intrinsic adenylate cyclase activity observed in bovine membranes for a series of NHH analogs with progressively diminishing affinity for receptors. The model represents an explicit theory of coupling where a successive series of temporal events are quantitatively related to each other and privide major constraints to any interpretation of the molecular organization of receptors and adenylate cyclase units in membranes. The model excludes a number of mechanistic proposals and suggests a new hypothesis for membrane coupling with features which may be generally applicable to other hormone-sensitive adenylate cyclase systems.


Three-dimensional image analysis of myosin head in function as captured by quick-freeze deep-etch replica electron microscopy.
Quick-freeze deep-etch replica electron microscopy combined with mica-flake technique provides high contrast, high time- and spatial-resolution images of protein molecules in solution, whose three-dimensional structure is well preserved. Thus, it might be quite useful to obtain structural information of individual functioning molecules, such as myosin crossbridges under in vitro motility assay conditions. With that method, we could actually show that both heads of heavy meromyosin (HMM) crossbridges are mostly straight and bound to actin filaments with about 45 degree tilt-angle under rigor conditions, whereas they attached to actin through only one head with a wide variety of angles under in vitro sliding conditions. We also demonstrated that free HMM heads are strongly kinked in the presence of ATP or ADP/inorganic vanadate (Vi) in contrast to almost straight configuration in the absence of nucleotide. To examine more detailed structure of individual crossbridges, we tried to reconstruct the three-dimensional architecture of intramolecular subdomains of single HMM molecule. We took a series of tilted images of single HMM-ADP/Vi particle and successfully obtained its 3-D image by filtered back-projection, even with restricted range of tilt-angles. By comparison of the reconstruction with the atomic model of subfragment-1 (S1) without nucleotide, we found some great structural difference, which partly might be attributable to the conformational change by nucleotide binding.


A functional variant of the collagen type III alpha1 gene modify risk of sporadic intracranial aneurysms.
Abnormalities in type III collagen in the arterial walls cause certain familial intracranial aneurysms (IAs); however, it remains unknown whether COL3A1 variants contribute to the risk of sporadic IAs. To study whether COL3A1 variants are associated with sporadic IAs, the association of COL3A1 variants with sporadic IAs was tested in 298 cases and 488 controls, replicated in an independent population of 192 cases and 1,690 controls, and further verified in 633 patients with intra-cerebral hemorrhage, 1,074 hypertensives, and 1,883 controls. We found that allele A of SNP rs1800255 conferred a 1.71-fold increased risk for IAs (adjusted odds ratio: OR = 1.71, 95% confidence interval: CI 1.19-2.45, P = 0.004) and results in an amino acid change of Ala698Thr, which led to a lower thermal stability of the peptide. These results were confirmed in the independent study. The associations were independent of the presence of hemorrhagic stroke and hypertension. These results support the view that the functional variant of COL3A1 is genetic risk factors for IAs in the Chinese population.


Should bleeding tendency deter abdominal paracentesis?
This study was conducted to evaluate the complications and bleeding associated with either thrombocytopoenia or prolongation of prothrombin time for ultrasound-guided abdominal paracentesis in the emergency department. In an emergency department of a tertiary centre, patients receiving ultrasound-guided abdominal paracentesis by the emergency physicians were prospectively enrolled. Patient characteristics, the preprocedure international normalised ratio for prothrombin time and the platelet count, and the procedure-related complications were collected and analysed. For a 2-year study period, a total of 410 abdominal paracenteses in 163 patients were investigated. The preprocedure international normalised ratio for prothrombin time was more than 1.5 in 142 paracenteses; the preprocedure platelet count was less than 50 x 10(3) microL(-1) in 55 paracenteses. Only two out of 410 procedures (0.5%, 95% confidence interval=0.1-1.8%) were associated with minor complications of cutaneous bleeding in the same patient (0.6%, 95% confidence interval=0.1-3.4%) at different visits. There was no significant procedure-related bleeding or complications even in patients with marked thrombocytopoenia or prolongation in international normalised ratio. Bleeding complication of ultrasound-guided abdominal paracentesis is uncommon and appears to be very mild, regardless of preprocedure international normalised ratio or platelet count. Routine correction of prolonged international normalised ratio or thrombocytopoenia before abdominal paracentesis may not be necessary.


On the distribution of [met5]- and [leu5]-enkephalins in the brain of the rat, guinea-pig and calf.
1 By means of a highly sensitive radioimmunoassay, the content of [met5]-enkephalin (ME) and [leu5]-enkephalin (LE) was measured in various regions of the rat, guinea-pig and calf brain. Provisions were made to differentiate ME from LE by the use of cyanogen bromide (CNBr) to destroy methionine, the carboxy terminal amino acid in the ME sequence. This allows for correction of possible errors due to the cross-reactivity of the ME to the LE antiserum. Evidence is presented to demonstrate that the specific radioimmunoassay combined with the CNBr technique is a valid method to measure LE and ME content in crude tissue extracts. 2 In all the species studied, enkephalins appeared to be highly concentrated in the striatum and hypothalamus while very low amounts were found in the cerebe-lum and hippocampus. 3 Although the ratio between ME and LE content varied from area to area, the ME content in every region of the rat, guinea-pig and calf brain was more than 4 fold greater than that of LE.


Clinical experiences with venetoclax and other pro-apoptotic agents in lymphoid malignancies: lessons from monotherapy and chemotherapy combination.
BH3-mimetics are a novel drug class of small molecule inhibitors of BCL2 family proteins which restore apoptosis in malignant cells. The only currently approved BH3-mimetic, the selective BCL2 inhibitor venetoclax, is highly efficacious in chronic lymphocytic leukemia and has rapidly advanced to an approved standard of care in frontline and relapsed disease in combination with anti-CD20 monoclonal antibodies. In this context, tumour lysis syndrome and myelosuppression are the most commonly encountered toxicities and are readily manageable with established protocols. Venetoclax is active in other lymphoid malignancies including several B cell non-Hodgkin lymphomas, acute lymphoblastic leukemia and multiple myeloma, with the highest intrinsic sensitivity observed in mantle cell lymphoma and Waldenstrom macroglobulinemia. Venetoclax combination with standard regimens in follicular lymphoma, multiple myeloma and aggressive B cell neoplasms has shown some promise, but further studies are required to optimize dose and scheduling to mitigate increased myelosuppression and infection risk, and to find validated biomarkers of venetoclax sensitivity. Future research will focus on overcoming venetoclax resistance, targeting other BCL2 family members and the rational design of synergistic combinations.


Morphological and biomechanical study of abdominal aorta of rats submitted to experimental chronic alcoholism.
To assess the influence of experimental chronic alcoholism in the morphology and biomechanics of the aorta, in a rat model. Forty-four Wistar rats were randomly divided into two groups of 22 animals each: in the alcoholism group, the rats received water mixed to increasing concentrations of ethyl alcohol; and control group: the rats received pure water for 180 days. The animals were then killed by an overdose of pentobarbital and the abdominal aortas were excised for histological (hematoxylin-eosin, Masson, Calleja, and Picrosirius red stain), histomorphometrical, and biomechanical analysis. Histology and histomorphometry did not show differences in aorta morphology of both groups. Biomechanical analysis showed a significantly greater yield point elongation in the alcoholic rat group (p < 0.05). Chronic alcoholism did not cause morphological alterations in the aortic wall but increased elongation, without modifying any other mechanical properties.


Engineering knottins as novel binding agents.
Cystine-knot miniproteins, also known as knottins, contain a conserved core of three tightly woven disulfide bonds which impart extraordinary thermal and proteolytic stability. Interspersed between their conserved cysteine residues are constrained loops that possess high levels of sequence diversity among knottin family members. Together these attributes make knottins promising molecular scaffolds for protein engineering and translational applications. While naturally occurring knottins have shown potential as both diagnostic agents and therapeutics, protein engineering is playing an important and increasing role in creating designer molecules that bind to a myriad of biomedical targets. Toward this goal, rational and combinatorial approaches have been used to engineer knottins with novel molecular recognition properties. Here, methods are described for creating and screening knottin libraries using yeast surface display and fluorescence-activated cell sorting. Protocols are also provided for producing knottins by synthetic and recombinant methods, and for measuring the binding affinity of knottins to target proteins expressed on the cell surface.


Retinoids and a retinoic acid receptor differentially modulate thymosin beta 10 gene expression in transfected neuroblastoma cells.
1. Investigations have demonstrated that the gene encoding thymosin beta 10 (a 43-amino acid member of a family of related proteins originally described in the rat immune system) is a target for morphogenic retinoids in both human and rat neuroblastoma cells. 2. Structure-activity studies revealed that the stimulatory actions of retinoids upon the thymosin beta 10 gene reflect the differing affinities of retinoid analogues for a retinoic acid receptor. 3. To examine further the possibility that the trophic actions of retinoic acid upon expression of the thymosin beta 10 gene involved retinoid receptors, neuroblastoma cells were transiently transfected with an expression vector encoding the nuclear retinoic acid receptor (alpha) protein. 4. Northern blot and slot-blot analyses revealed that neuronal cells overexpressing RAR alpha-mRNA exhibited an enhanced sensitivity to exogenous and endogenous retinoic acid in terms of thymosin beta 10 mRNA. Although the RAR-alpha gene was expressed (at low levels) a priori in these neuroblastoma cells, retinoic acid (2 x 10(-7) M for 3 days) slightly stimulated RAR-alpha-mRNA accumulation. 5. Collectively, these findings indicate the retinoic acid receptor (alpha) is regulated by retinoid acid and that the developmentally regulated, retinoid-responsive thymosin beta 10 gene is a target for this nuclear transcription factor in cells derived from the neural crest.


Safety effects of the separation distances between driveway exits and downstream U-turn locations.
Using U-turns as alternatives to direct left-turns is an important access management treatment which has been widely implemented in the United States to improve safety on multilane highways. The primary objective of this study is to evaluate the safety effects of the separation distances between driveway exits and downstream U-turn locations. To achieve the research objective, crash data reported at 140 street segments in the state of Florida were investigated. The selected sites were divided into three groups based on the separation distances. t-Tests and proportionality tests were performed for comparing crash frequency, crash type, and crash severity between different separation distance groups. Negative-binomial models were developed for examining the factors that contribute to the crashes reported at selected sites. The data analysis results show that the separation distances significantly impact the safety of the street segments between driveways and downstream U-turn locations. A 10% increase in separation distance will result in a 3.3% decrease in total crashes and a 4.5% decrease in the crashes which is related with right-turns followed by U-turns. The models also show that providing U-turns at a signalized intersection will result in more crashes at weaving sections. Thus, if U-turns are to be provided at a signalized intersection, a longer separation distance shall be provided.


Alzheimer's disease--a dysfunction in cholesterol and lipid metabolism.
1. Strong etiological association exists between dysfunctional metabolism of brain lipids, age-related changes in the cerebral vasculature and neurodegenerative features characteristic of Alzheimer's disease (AD) brain. 2. In this short review, recent experimental evidence for these associations is further discussed below.


Toxic response indicators of microcystin-LR in F344 rats following a single-dose treatment.
Microcystin-LR (MCLR) is the most common hepatotoxic cyanotoxin produced primarily by Microcystis aeruginosa. In this study, young F344 rats were intraperitoneally injected with a single dose (25, 50, 100, and 150 microg/kg) of MCLR to explore possible toxic effect and toxic response indicators. Acute toxic symptoms, including body weight loss and death, were monitored for 7 days. Mortality reached 100% (9/9) in rats treated with a single MCLR dose of 150 microg/kg. Histopathological examination showed spot necrosis in the liver of animals treated at low doses, while massive hemorrhage and widespread necrotic foci occurred at higher doses, indicating extensive liver damage. Protein phosphatase 2A (PP2A) expression showed a dose-dependent decrease in the liver. Immunohistochemical localization indicated that nuclear PP2A was affected first, followed by cytoplasmic PP2A. In addition, there was a significant increase in sphingolipid levels at higher doses, indicating the involvement of a ceramide-mediated apoptotic pathway. Expression of apoptosis and cell cycle regulatory proteins like Bax, Bcl2, and Bad showed a dose-dependent decrease. This study demonstrated that treatment with a single dose of MCLR caused liver damage, increased sphingolipid levels, and decreased PP2A expression, which ultimately down-regulated the expression of Bcl2 family proteins.


Cofilin (ADF) affects lateral contacts in F-actin.
The effect of yeast cofilin on lateral contacts between protomers of yeast and skeletal muscle actin filaments was examined in solution. These contacts are presumably stabilized by the interactions of loop 262-274 of one protomer with two other protomers on the opposite strand in F-actin. Cofilin inhibited several-fold the rate of interstrand disulfide cross-linking between Cys265 and Cys374 in yeast S265C mutant F-actin, but enhanced excimer formation between pyrene probes attached to these cysteine residues. The possibility that these effects are due to a translocation of the C terminus of actin by cofilin was ruled out by measurements of fluorescence resonance energy transfer (FRET) from tryptophan residues and ATP to acceptor probes at Cys374. Such measurements did not reveal cofilin-induced changes in FRET efficiency, suggesting that changes in Cys265-Cys374 cross-linking and excimer formation stem from the perturbation of loop 262-274 by cofilin. Changes in lateral interactions in F-actin were indicated also by the cofilin-induced partial release of rhodamine phalloidin. Disulfide cross-linking of S265C yeast F-actin inhibited strongly and reversibly the release of rhodamine phalloidin by cofilin. Overall, this study provides solution evidence for the weakening of lateral interactions in F-actin by cofilin.


Reduction of food intake following X-ray irradiation of rats--involvement of visceral afferent nerves.
Radiotherapy for malignant tumours often elicits anorexia or loss of appetite as an adverse effect. However, the mechanism for this is poorly understood. The present study was undertaken to investigate if visceral afferents are responsible for reduction of food intake following X-ray irradiation. Rats were exposed bilaterally to X-ray (10 MV) irradiation with total doses of 1.5, 3 and 6 Gy, using a high-energy electron linear accelerator at a dose rate of 4.9 Gy min(-1) X-ray irradiation of the whole body, abdomen or head with doses of 1.5, 3 and 6 Gy reduced food intake in a dose-dependent manner. The reduction of food intake after X-ray irradiation of the whole body or abdomen was significantly greater than when only the head was irradiated. Reduction of food intake was observed for the first 4 days after 6 Gy X-ray irradiation of the abdomen, while it was observed only on the first day after the same 6-Gy irradiation of the abdomen in animals whose small-diameter afferents were ablated by capsaicin pre-treatment. These results suggested that the abdominal afferent nerves at least contribute to the reduction of food intake observed on second to fourth days after 6-Gy abdominal irradiation. Taken together, the present evidence suggests that the reduction of food intake following X-ray irradiation of the whole body or the abdomen is partly mediated via abdominal afferent nerves. Moreover, the results of X-ray irradiation to the head suggest that X-ray irradiation directly influences the central nervous system to reduce food intake.


Efficacy of Adiantum capillus veneris Linn in chemically induced urolithiasis in rats.
Adiantum capillus veneris Linn has been recommended in ancient literature of Unani system of medicine as an important ingredient of many formulations for the treatment of urolithiasis. Its decoction has long been used for the same purpose by several Unani physicians. To investigate the antiurolithiasic effect of the hydro alcoholic extract of Adiantum capillus veneris Linn in male Sprague Dawley rats. The effects of oral administration of hydro alcoholic extract of test drug were studied on calcium oxalate urolithiasis. A total of 48 rats were used for the study. The animals were divided into six groups of eight animals each. Plain control rats were treated with distilled water only, throughout the study period, whereas in other groups nephrolithiasis was induced by providing drinking water containing 0.75% ethylene glycol and 1% ammonium chloride for 7 days. Thereafter, urine was examined for the presence of crystals. Negative control group A rats were sacrificed after 7 days, whereas negative control group B was left untreated up to the end of study. Test groups were treated with 127.6 mg/kg and 255.2 mg/kg of test drug and standard control with Cystone (750 mg/kg) for 21 days. At the end of experiment, number of crystals in urine and levels of calcium, phosphorus, urea and creatinine in serum were observed. Histopathological study of the kidney was done by light microscopy. Urine microscopy showed significant reduction (p<0.001 and p<0.01) in the number of crystals in test groups A and B respectively. Serum levels of calcium, phosphorous, and blood urea were found to be decreased significantly in all the groups. In both the test groups, serum creatinine level was found to be similar as in plain control. The animals treated with test drug showed much improvement in body weight. Histopathology of kidney showed almost normal kidney architecture in treated groups. The above findings indicate the antiurolithic activity of Adiantum capillus veneris Linn, and thus, validate the claims of Unani physicians for its medicinal use in urolithiasis.


Polydatin protects cardiac function against burn injury by inhibiting sarcoplasmic reticulum Ca2+ leak by reducing oxidative modification of ryanodine receptors.
Our recent studies demonstrate that burn trauma induces leaky sarcoplasmic reticulum (SR) in heart due to excessively active ryanodine receptor (RyR) function. SR Ca(2+) leak causes partial depletion of SR Ca(2+) content and disturbances in intracellular Ca(2+) homeostasis, resulting in the pathogenesis of burn-generated cardiac dysfunction. This study investigated the role of polydatin, a resveratrol glucoside, in preventing SR leak and its therapeutic effect against burn-generated cardiac dysfunction. We found that polydatin treatment improved cardiac function impaired by burn injury of 30% of total body surface area. Parallel to the alterations in cardiac function, polydatin significantly increased the defective systolic Ca(2+) transient and contractility in burn-traumatized cardiomyocytes. Burn injury increased the occurrence of Ca(2+) sparks. The enhancement of Ca(2+) spark-mediated SR leak caused partial depletion of SR Ca(2+) content in burn-traumatized cardiomyocytes. Furthermore, we found that the content of free thiols (the number of reduced cysteines) in RyR2 in cardiomyocytes determined by the monobromobimane fluorescence of RyR2 was decreased markedly in burn-traumatized hearts. Polydatin treatment decreased intracellular reactive oxygen species levels and restored the amount of free thiols in RyR2 in burns. Concomitantly, polydatin corrected Ca(2+) spark-mediated SR leak and restored SR Ca(2+) load. The systolic Ca(2+) transient and cellular contractility were significantly increased by polydatin treatment. Taken together, the present findings provide the first evidence demonstrating that polydatin prevents enhanced Ca(2+) spark-mediated SR leak by reducing oxidative stress in RyR2 in burn-traumatized heart, leading to protection of cardiac function against burn injury.


Investigation of uranium-colloid interactions in soil by dual field-flow fractionation/capillary electrophoresis hyphenated with inductively coupled plasma-mass spectrometry.
This paper deals with the study of uranium-colloid interactions in a carbonated soil. The work is focused on the immediately available fraction obtained after a leaching process, according to a normalized batch method. In order to characterize the different colloidal carriers, Asymmetrical Flow Field-Flow Fractionation (As-Fl-FFF) coupled to different detectors (UV, Multi Angle Laser Light Scattering (MALLS) and Inductively coupled Plasma-Mass Spectrometry (ICP-MS)) was used. The colloidal carriers are mainly inorganic particles (carbonated particles and clays) mixed with organic substances. Furthermore, dissolved and colloidal uranium species in the leaching solutions were monitored by Capillary Electrophoresis (CE) coupled to ICP-MS, in order to investigate the uranium/colloids interactions. According to the first results, uranium fate in this specific soil is controlled by sorption/desorption phenomena, strongly pH dependent.


Simultaneous administration of multiple model substrates to assess hepatic drug clearance.
We have evaluated a method to simultaneously assess three major processes involved in hepatic drug clearance using three model substrates administered simultaneously as a 5-minute intravenous injection. Lorazepam, indocyanine green, and antipyrine are used to assess conjugation, liver blood flow, and microsomal oxidative metabolism, respectively. These substrates were administered individually and as a mixture to 10 healthy adult male volunteers to determine if clearances of any of the compounds were affected by simultaneous administration. Mean clearances of the substrates were not different when administered alone (9.97, 0.78, and 0.53 ml/min/kg) vs. together (11.5, 0.89, and 0.52 ml/min/kg), using a paired t test. Since we were using this method to assess hepatic drug clearance in children with leukemia, the effect of short-term allopurinol was assessed. The three model substrates were administered to the volunteers after 0, 1, 8, and 22 days of treatment with allopurinol, 200 mg t.i.d. There was no change in mean clearance of any of the three compounds at any point during allopurinol treatment (repeated-measures ANOVA). We conclude that this technique is a simple and valid method to simultaneously assess three major processes involved in hepatic drug clearance and is not affected by up to 22 days of oral allopurinol treatment. This simple technique, requiring a single set of blood samples, has potential applications in the assessment of developmental changes in hepatic drug clearance, as well as the effects of environmental, therapeutic, and pathophysiologic factors on three major processes involved in hepatic drug clearance.


RADON EXPOSURE AT SPECIFIC WORKPLACES IN AUSTRIA-EXPERIENCES AND FUTURE CHALLENGES.
Radon exposure at specific workplaces like water works, radon spas, tourist mines and show caves were evaluated in Austria within pilot studies (2004-10) and have been regulated in an ordinance since 2008. In this article, the major results and experiences in radon measurements and dose assessments at those workplaces by the accredited AGES laboratory are presented.


The unequivocal determination of lactic acid using a one-dimensional zero-quantum coherence-transfer technique.
A method based on zero-quantum coherence transfer for spectral editing of metabolites in aqueous solution in a one-dimensional experiment is described. Water suppression factors of approximately 8000 were achieved by the use of pulsed B0 field gradients and excellent editing was obtained in a single acquisition. The methyl resonances of both lactate and acetaldehyde were readily observed in a mouse brain homogenate by generating zero-quantum coherence using Gaussian pulses for selective excitation of the respective CH resonances.


Local immunosuppression in vascularized composite allotransplantation (VCA): A systematic review.
Local immunosuppression in vascularized composite allotransplantation (VCA) aims to minimize immunosuppressant-related toxic and malignant side effects. Promising allograft survival data have been published by multiple workgroups. In this systematic review, we examine preclinical animal studies that investigated local immunosuppression in VCA. We conducted a systematic review of manuscripts listed in the MEDLINE and PubMed database concerning preclinical VCA models. Papers included had to be available as full-text and written in English. Non-VCA studies, human trials, and studies using cell-based therapy strategies were excluded. We followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Literature research retrieved 980 articles. Ten studies published between 2010 and 2019 met the inclusion and exclusion criteria. Seven out of ten articles demonstrated a significant prolongation of allograft survival by using local immunosuppression. Five articles employed tacrolimus (TAC) as the main immunosuppressive agent. Seven studies performed hind-limb VCA in a rat model. The easily accessible location of skin containing VCAs makes it an ideal candidate for local immunosuppression. Published preclinical data are very promising in terms of improved allograft survival and reduced systemic toxicity.


Involvement of phospholipase D in store-operated calcium influx in vascular smooth muscle cells.
In non-excitable cells, sustained intracellular Ca2+ increase critically depends on influx of extracellular Ca2+. Such Ca2+ influx is thought to occur by a 'store-operated' mechanism, i.e. the signal for Ca2+ entry is believed to result from the initial release of Ca2+ from inositol 1,4,5-trisphosphate-sensitive intracellular stores. Here we show that the depletion of cellular Ca2+ stores by thapsigargin or bradykinin is functionally linked to a phosphoinositide-specific phospholipase D (PLD) activity in cultured vascular smooth muscle cells (VSMC), and that phosphatidic acid formed via PLD enhances sustained calcium entry in this cell type. These results suggest a regulatory role for PLD in store-operated Ca2+ entry in VSMC.


Modified Nephrometry Score With Body Mass Index More Accurately Predicts Ischemic Time in Transabdominal Laparoscopic Partial Nephrectomy for Small Renal Masses.
To accurately predict the ischemic time (IT) and select candidates for transabdominal laparoscopic partial nephrectomy (LPN). Transabdominal LPN was performed for 135 Japanese patients with renal masses <7 cm in diameter between 2009 and 2016 by a single surgeon in a single institute. The renal parenchymal sutures were done with a combination of felt, Hem-o-lok and Lapra-Ty. The original R.E.N.A.L. nephreometry score (NS) was modified as follows. In terms of the tumor size (R), 1, 2, and 3 points were given for ≤2.5, 2.5-4, and >4 cm, respectively. For tumor location (A), 1, 2, and 3 points were given for anterior, on the coronal plane, and posterior, respectively. The median IT was 24.6 minutes. Using the original NS, 64, 65, and 6 patients were categorized into low (4-6), moderate (7-9), and high (10-12) complexity, respectively. With the modified NS, 42, 75, and 18 patients were categorized into low (5-7), moderate (8-11), and high (12-15) complexity, respectively. Pearson's correlation coefficient (R) between the original NS and IT was 0.297, whereas the R was improved to 0.388 when the modified NS was employed. If the modified NS was low complexity, 95.2% achieved IT <30 minutes. Of them, none showed IT ≥30 minutes if the body mass index was <25 kg/m2. The modified NS had a better correlation to the IT than the original NS for patients with tumor sizes <7 cm who underwent transabdominal LPN. The modified NS with body mass index is a practical tool to select candidates for transabdominal LPN.


Site-directed mutagenesis of HLA molecules reveals the functional epitope of a human HLA-A1/A36-specific monoclonal antibody.
Eplet 44KM is currently listed in the HLA Epitope Registry but does not adhere to the eplet definition of an amino acid configuration within a 3.5 Å radius. Eplet 44KM has been previously redefined to the antibody-verified reactivity pattern 44K/150V/158V, based on reactivity analysis of monoclonal antibody VDK1D12. Since the three residues are always simultaneously present on common HLA alleles, methods to define which residue is crucial for antibody-induction and binding are limited. In this proof-of-concept study, we performed site-directed mutagenesis to narrow down the antibody-verified reactivity pattern 44K/150V/158V to a single amino acid and defined 44K as the eplet or functional epitope of mAb VDK1D12.


On the cause of spatial displacement of long T1 species in segmented inversion recovery prepared imaging.
Using inversion recovery steady-state free precession segmented k-space imaging for the detection of myocardial infarction, we noticed that some structures appeared in the wrong locations of the image. In this work, the spatial displacement is demonstrated and explained from both theoretical and experimental points of view. The effect is due to a change in phase from segment to segment of the detected magnetization from species with long T1's such as cysts, fluid collections, and cerebrospinal fluid. Depending on the number of k-space segments and view ordering, structures can be replicated throughout the image or displaced by half of the phase-encoding field of view.


A prognostic signature of defective p53-dependent G1 checkpoint function in melanoma cell lines.
Melanoma cell lines and normal human melanocytes (NHM) were assayed for p53-dependent G1 checkpoint response to ionizing radiation (IR)-induced DNA damage. Sixty-six percent of melanoma cell lines displayed a defective G1 checkpoint. Checkpoint function was correlated with sensitivity to IR with checkpoint-defective lines being radio-resistant. Microarray analysis identified 316 probes whose expression was correlated with G1 checkpoint function in melanoma lines (P≤0.007) including p53 transactivation targets CDKN1A, DDB2, and RRM2B. The 316 probe list predicted G1 checkpoint function of the melanoma lines with 86% accuracy using a binary analysis and 91% accuracy using a continuous analysis. When applied to microarray data from primary melanomas, the 316 probe list was prognostic of 4-yr distant metastasis-free survival. Thus, p53 function, radio-sensitivity, and metastatic spread may be estimated in melanomas from a signature of gene expression.


Prevalence of rib anomalies in normal Caucasian children and childhood cancer patients.
To evaluate the prevalence of abnormalities of rib development in normal Caucasian children and patients with childhood cancer. Chest radiographs of 881 Caucasian pediatric controls and 906 childhood cancer patients were reviewed, and independently scored by four blinded observers, using strict definitions. Prevalences of 6 major rib anomaly categories in controls were compared to their prevalence in the total group of childhood cancer patients, and the 12 individual larger tumor groups using Chi-square tests. Values in the control population were generated for the occurrence of six major rib anomaly categories; cervical rib anomalies were present in 6.1% of controls, aplasia of 12th ribs in 6.6%, lumbar ribs in 0.9%, bifurcations in 0.7%, synostosis-bridging in 0.3%, and segmentations were not found. The overall prevalence of total rib anomalies in cases and controls was equal (14.9% and 14.2%, respectively). Cervical rib anomalies were found significantly more often in cases (8.6%) compared to controls (p-value=0.047), three groups accounting for this higher prevalence: 12.1% of acute lymphoblastic leukemia patients (p=0.011), 18.2% of astrocytoma patients (p=0.023), and 14.7% of germ cell tumor patients (p=0.046) had a cervical rib anomaly. Prevalence figures for the presence and type of rib anomalies in a large group of normal Caucasian children were generated. In childhood cancer patients a significantly higher prevalence of cervical rib anomalies was demonstrated in patients with acute lymphoblastic leukemia, astrocytoma, and germ cell tumors.


Genomic regulation after CD40 stimulation in microglia: relevance to Alzheimer's disease.
Key pathological processes in Alzheimer's disease (AD) include the accumulation of amyloid beta peptide (Abeta) which, in excess, triggers pathological cascades including widespread inflammation, partly reflected by chronic microglial activation. It has previously been suggested that CD40/CD40L interaction promotes AD like pathology in transgenic mice. Thus, amyloid burden, gliosis and hyperphosphorylation of tau are all reduced in transgenic models of AD lacking functional CD40L. We therefore hypothesized that cellular events leading to altered APP metabolism, inflammation and increased tau phosphorylation underlying these observations would be regulated at the genomic level. In the present report, we used the Affymetrix (GeneChip) oligonucleotide microarray U133A to gain insight into the global and simultaneous transcriptomic changes in response to microglia activation after CD40/CD40L ligation. As expected, regulation of elements of the NF-kappaB signaling, chemokine and B cell signaling pathways was observed. Taken together, our data also suggest that CD40 ligation in human microglia specifically perturbs many genes associated with APP processing.


Saethre-Chotzen syndrome: long-term outcome of a syndrome-specific management protocol.
To assess the long-term outcomes of our management protocol for Saethre-Chotzen syndrome, which includes one-stage fronto-orbital advancement. All patients born with Saethre-Chotzen syndrome between January 1992 and March 2017 were included. Evaluated parameters included occipital frontal head circumference (OFC), fundoscopy, neuroimaging (ventricular size, tonsillar position, and the presence of collaterals/an abnormal transverse sinus), polysomnography, and ophthalmological outcomes. The relationship between papilledema and its associated risk factors was evaluated with Fisher's exact test. Thirty-two patients (21 females, 11 males) were included. Median (SD) age at first surgery was 9.6 months (3.1mo) for patients who were primarily referred to our center (range: 3.6-13.0mo), the median (SD) age at last follow-up was 13 years (5y 7mo; range: 3-25y). Seven patients had papilledema preoperatively, which recurred in two. Two patients had papilledema solely after first surgery. Second cranial vault expansion was indicated in 20%. Thirteen patients had an OFC deflection, indicating restricted skull growth, one patient had ventriculomegaly, and none developed hydrocephalus. Eleven patients had emissary veins, while the transverse sinus was aberrant unilaterally in 13 (hypoplastic n=10 and absent n=3). Four patients had mild tonsillar descent, one of which was a Chiari type I malformation. Four patients had obstructive sleep apnoea (two mild, one moderate, and one severe). An aberrant transverse sinus was associated with papilledema (p=0.01). Single one-stage fronto-orbital advancement was sufficient to prevent intracranial hypertension for 80% of our patients with Saethre-Chotzen syndrome. Follow-up should focus on OFC deflection and venous anomalies.


HIV in pregnancy.
Human immunodeficiency virus is becoming increasingly common among childbearing women. Through federally recommended testing programs, obstetricians will soon be identifying pregnant women in their practices who are HIV infected. Hence obstetricians must be prepared to face the social, ethical, and medical dilemmas brought on by HIV infection in pregnancy.


Cropland expansion in the United States produces marginal yields at high costs to wildlife.
Recent expansion of croplands in the United States has caused widespread conversion of grasslands and other ecosystems with largely unknown consequences for agricultural production and the environment. Here we assess annual land use change 2008-16 and its impacts on crop yields and wildlife habitat. We find that croplands have expanded at a rate of over one million acres per year, and that 69.5% of new cropland areas produced yields below the national average, with a mean yield deficit of 6.5%. Observed conversion infringed upon high-quality habitat that, relative to unconverted land, had provided over three times higher milkweed stem densities in the Monarch butterfly Midwest summer breeding range and 37% more nesting opportunities per acre for waterfowl in the Prairie Pothole Region of the Northern Great Plains. Our findings demonstrate a pervasive pattern of encroachment into areas that are increasingly marginal for production, but highly significant for wildlife, and suggest that such tradeoffs may be further amplified by future cropland expansion.


The distribution of the orphan bombesin receptor subtype-3 in the rat CNS.
Bombesin receptor subtype 3 (BRS-3) is an orphan G-protein coupled receptor that shares between 47 and 51% homology with other known bombesin receptors. The natural ligand for BRS-3 is currently unknown and little is known about the mechanisms regulating BRS-3 gene expression. Unlike other mammalian bombesin receptors that have been shown to be predominantly expressed in the CNS and gastrointestinal tract, expression of the BRS-3 receptor in the rat brain has previously not been observed. To gain further understanding of the biology of BRS-3, we have studied the distribution of BRS-3 mRNA and protein in the rat CNS. The mRNA expression pattern was studied using reverse transcription followed by quantitative polymerase chain reaction. Using immunohistological techniques, the distribution of BRS-3 protein in the rat brain was investigated using a rabbit affinity-purified polyclonal antiserum raised against an N-terminal peptide. The BRS-3 receptor was found to be widely expressed in the rat brain at both mRNA and protein levels. Particularly strong immunosignals were observed in the cerebral cortex, hippocampal formation, hypothalamus and thalamus. Other regions of the brain such as the basal ganglia, midbrain and reticular formation were also immunopositive for BRS-3. In conclusion, our neuroanatomical data provide evidence that BRS-3 is as widely expressed in the rat brain as other bombesin-like peptide receptors and suggest that this receptor may also have important roles in the CNS, mediating the functions of a so far unidentified ligand.


Site specific incidence rate of genomic subtypes of enteropathogenic Escherichia coli and association with enteric inflammation and child growth.
There is a lack of information highlighting the possible association between the genomic subtypes of enteropathogenic Escherichia coli (EPEC) on environmental enteric dysfunction (EED) and on linear growth during childhood. Genomic subtypes of EPEC from stool samples collected from 1705 children enrolled in the MAL-ED birth cohort were detected by TaqMan Array Cards. We measured site-specific incidence rate by using Poisson regression models, identified the risk factors and estimated the association of genomic subtypes of EPEC with the composite EED score and linear growth at 24 months of age. In general, the highest incidence rate (39%) was found among children having aEPEC infection, which was the greatest in Tanzania (54%). Exclusive breastfeeding and having an improved sanitation facility were found to be protective factors against EPEC infection. In the multivariate models, in overall effect after adjusting for the potential covariates aEPEC showed strong positive associations with the EED scores and tEPEC showed a positive association with poor linear growth at 24 months of age. Our analyses may lay the cornerstone for a prospective epidemiologic investigation for a potential vaccine development aimed at reducing the burden of EPEC infections and combat childhood malnutrition.


Effect of malnutrition during the first year of life on adult plasma insulin and glucose tolerance.
There is evidence linking intrauterine growth retardation with increased cardiovascular risk and diabetes mellitus (DM) later in life. However, little is known about the association between malnutrition during the first year of life and metabolic abnormalities in adulthood. The objective of this study was to assess the effect of documented malnutrition during the first year of life on glucose tolerance, plasma insulin, lipid profile, and blood pressure in early adulthood, as well as to assess the interaction between body mass index (BMI) and malnutrition on these variables. A study group of young men with a documented history of malnutrition during their first year of life was recruited from 4 pediatric hospitals in Mexico City and compared with a control group. Subjects included were 52 men, aged 20.2 +/- 3.6 years, with a mean birth weight of 3.0 +/- 0.7 kg and documented malnutrition in their first year of life; controls were 50 men, aged 23.3 +/- 1.8 years, with a mean birth weight of 3.2 +/- 0.5 kg. Insulin and glucose concentrations, fasting and in response to an oral glucose load, plasma lipids, blood pressure, and an insulin sensitivity index (ISI) were measured. The areas under the curves of glucose (AUCG) and insulin (AUCI) were significantly higher in cases (P =.012 and <.002, respectively), independent of birth weight, BMI, or age. BMI was significantly associated with fasting plasma insulin (FPI), AUCI, ISI, triglyceride, and high-density lipoprotein (HDL)-cholesterol concentrations in cases, but not in controls. These data suggest that early malnutrition in extrauterine life, independently of birth weight, has an adverse effect on insulin metabolism and glucose tolerance in young men, and it worsens as body mass increases even within the normal range of BMI. Therefore, it is advisable to prevent obesity in individuals exposed to early malnutrition.


Analysis of the repertoire of human B-lymphocytes specific for type A and type B blood group terminal trisaccharide epitopes.
Naturally occurring serum antibodies specific for the A and B blood group isoantigens are of great importance in medicine. By using A-type terminal trisaccharide (ATS) or B-type terminal trisaccharide (BTS) coupled to albumin as coating antigens, an enzyme-linked immunosorbent assay capable of detecting all ATS/BTS-binding antibodies was performed. The combination of this enzyme-linked immunosorbent assay with limiting-dilution methodology, using a polyclonal B-lymphocyte activator, permitted a monoclonal analysis of the human antibody repertoire that is specific for ATS and BTS in persons of different blood types. Most (78%) positive supernatants from type O cultures were monospecific for either ATS or BTS, and these were present at roughly equivalent frequencies. Nine supernatants with reactivity toward both ATS and BTS were tested by red cell adsorption; six had properties expected for true dually reactive monoclonal antibodies: adsorption with either A1 or B red cells eliminated both anti-ATS and anti-BTS activity. This finding accords with a monoclonal origin for anti-A,B. The analysis of cultures of peripheral blood lymphocytes from type A and B donors unexpectedly showed significant numbers of clones with apparent autospecificity. However, none of the anti-ATS-positive supernatants from type A cultures or anti-BTS-positive supernatants from type B cultures were adsorbable with A1 or B red cells, respectively. Consideration of only true (adsorbable) positives indicates that the type A and B anti-trisaccharide repertoires differ significantly from the type O repertoire, probably as a result of the action of normal self-tolerance mechanisms.


Measurement of ATP turnover during shortening and lengthening of rabbit psoas myofibrils using a fluorescent ATP analog.
In order to study ATP turnover during shortening and lengthening of rabbit psoas myofibrils, we have used fluorescence microscopy in which the displacement of a fluorescent nucleotide analog, 2'(3')-O-[N-[2-[[Cy3] amido] ethyl] carbamoyl]-adenosine 5' triphosphate (Cy3-EDA-ATP) bound to cross-bridge on flash photolysis of caged ATP was measured [Chaen et al. (1997) Biophys. J. 73, 2033-2042]. In the previous paper, we reported that when a myofibril was imposed to shorten with a constant velocity by a piezo-electric actuator, the nucleotide displacement rate constant initially increased to 0.7 s-1 with increasing shortening velocity and then declined with a further increase in shortening velocity. The rate constant during lengthening measured in the present experiment was found to be not significantly affected. These results suggest that the cross-bridge kinetics show a asymmetrical dependence on the mechanical strain in the cross-bridges, namely, the rate constants are not significantly affected at higher strain during lengthening but depend on the lower strain during shortening.


Are acute sitting-induced changes in inflammation and cerebrovascular function related to impaired mood and cognition?
Sedentary behaviour is negatively associated with mood and cognition, yet how acute sitting contributes to these overall associations is unknown. Since sitting heightens inflammation and impairs cerebrovascular function, this study investigated the hypothesis that these sitting-induced changes are related to impaired mood and cognition. Twenty-five healthy desk workers (18 male, 28.3 ± 7.5 years, BMI: 24.2 ± 3.3 kg∙m-2) were recruited. During laboratory visit one, participants were familiarised with cognitive performance tests measuring executive function, attention and working memory. During laboratory visit two, participants completed 6 h of continuous, uninterrupted sitting. At baseline and after 6 h, serum markers of inflammation, middle cerebral artery blood flow velocity (MCAv), cerebrovascular carbon dioxide reactivity (CVR), dynamic cerebral autoregulation (CA), cognitive performance and mood (positive and negative affect, alert, contented and calm) were assessed. Data were analysed using paired-samples t tests and correlation analyses. Following sitting, C-reactive protein (∆-1.0 µg/ml) and tissue plasminogen activator (∆-360.4 pg/ml) decreased (p < 0.05), MCAv reduced (∆-2.9 cm∙s-1, p = 0.012) and normalised gain increased in the very low frequency range, indicating impaired CA (∆ + 0.22%·mmHg-1, p = 0.016). Positive affect (∆-4.6, p < 0.001), and alert (∆-10.6 p = 0.002) and contented (∆-7.4, p = 0.006) mood states also decreased following sitting. No significant changes in interleukin-6, tumour necrosis factor-alpha, von Willebrand factor, CVR or cognitive performance were observed (p > 0.05). The observed changes in inflammation and cerebrovascular function were not related to changes in mood (p > 0.05). Alterations in inflammation or cerebrovascular function following six hours of prolonged, uninterrupted sitting are not related to the observed reductions in mood, indicating other mechanisms underlie the relationship between acute sitting and mood disturbances.


Potent triazolyl-proline-based inhibitors of HCV NS3 protease.
The design and synthesis of tripeptide-based inhibitors of the HCV NS3 protease containing a novel P2-triazole is described. Replacement of the P2 quinoline with a triazole moiety provided a versatile handle which could be expediently modified to generate a diverse series of inhibitors. Further refinement by the incorporation of an aryl-substituted triazole and replacement of the P1 acid with an acyl sulfonamide ultimately provided inhibitors with interesting cellular activity.


The augmentation of human NK-cell activity by auranofin compared to interferon.
The cytotoxic activity of natural killer cells is markedly increased by interferon (IFN) and IFN inducers. Auranofin, an oral gold preparation, has an effect similar in size and rapidity to that of IFN but, in contrast, the Auranofin effect does not require RNA synthesis by the effector cells.


Coupling gold nanoparticles to silica nanoparticles through disulfide bonds for glutathione detection.
Advances in the controlled assembly of nanoscale building blocks have resulted in functional devices which can find applications in electronics, biomedical imaging, drug delivery etc. In this study, novel covalent nanohybrid materials based upon [Ru(bpy)3](2+)-doped silica nanoparticles (SiNPs) and gold nanoparticles (AuNPs), which could be conditioned as OFF-ON probes for glutathione (GSH) detection, were designed and assembled in sequence, with the disulfide bonds as the bridging elements. The structural and optical properties of the nanohybrid architectures were characterized using transmission electron microscopy, UV-vis spectroscopy and fluorescence spectroscopy, respectively. Zeta potential measurements, x-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy were employed to monitor the reaction processes of the SiNPs-S-S-COOH and SiNPs-S-S-AuNPs synthesis. It was found that the covalent nanohybrid architectures were fluorescently dark (OFF state), indicating that SiNPs were effectively quenched by AuNPs. The fluorescence of the OFF-ON probe was resumed (ON state) when the bridge of the disulfide bond was cleaved by reducing reagents such as GSH. This work provides a new platform and strategy for GSH detection using covalent nanohybrid materials.


Utility of hemodialysis in maple syrup urine disease.
Maple syrup urine disease (MSUD) is an inborn error of metabolism stemming from a deficiency in 2-ketoacid dehydrogenase and resulting in the systemic accumulation of branched chain amino acids (BCAAs). Affected children may suffer profound developmental and cognitive impairment from exposure to high levels of BCAA and their associated neurotoxic metabolites. Endogenous renal clearance of BCAA is limited and several therapeutic modalities including intensive nutritional regimens, exchange transfusions, peritoneal dialysis, and continuous hemofiltration have been utilized in neonates with MSUD, all of which have had varying success in reducing systemic BCAA levels. In this report, a symptomatic 7-day-old 3-kg neonate with MSUD underwent treatment with a combination of early hemodialysis and aggressive enteral feedings of a metabolically appropriate formula. This approach results in a 75% reduction of systemic toxin levels within 3 h. When compared to other reported modalities of therapy for symptomatic neonates with MSUD, this approach appears to be most efficacious. Moreover, by minimizing the amount of time that an affected neonate is exposed to neurotoxic levels of BCAAs, long-term developmental and cognitive capabilities may be preserved.


Pride in STEM worldwide.
Cell asked LGBTQ+ scientists around the world about how their identity shapes their experiences in STEM. Here we share six unique perspectives of researchers highlighting how their area of expertise, research focus, institutions, and geographical location have played a role in this regard. We thank them for sharing their voices and continued efforts toward making science more inclusive.


Corrigendum to "Infection-related failure of autologous versus allogenic cranioplasty after decompressive hemicraniectomy - A systematic review and meta-analysis" [Brain Spine 3 (2023) 101760].
[This corrects the article DOI: 10.1016/j.bas.2023.101760.].


Enantioselective Rhodium-Catalyzed Coupling of Arylboronic Acids, 1,3-Enynes, and Imines by Alkenyl-to-Allyl 1,4-Rhodium(I) Migration.
A chiral rhodium complex catalyzes the highly enantioselective coupling of arylboronic acids, 1,3-enynes, and imines to give homoallylic sulfamates. The key step is the generation of allylrhodium(I) species by alkenyl-to-allyl 1,4-rhodium(I) migration.


Induced fit with replica exchange improves protein complex structure prediction.
Despite the progress in prediction of protein complexes over the last decade, recent blind protein complex structure prediction challenges revealed limited success rates (less than 20% models with DockQ score > 0.4) on targets that exhibit significant conformational change upon binding. To overcome limitations in capturing backbone motions, we developed a new, aggressive sampling method that incorporates temperature replica exchange Monte Carlo (T-REMC) and conformational sampling techniques within docking protocols in Rosetta. Our method, ReplicaDock 2.0, mimics induced-fit mechanism of protein binding to sample backbone motions across putative interface residues on-the-fly, thereby recapitulating binding-partner induced conformational changes. Furthermore, ReplicaDock 2.0 clocks in at 150-500 CPU hours per target (protein-size dependent); a runtime that is significantly faster than Molecular Dynamics based approaches. For a benchmark set of 88 proteins with moderate to high flexibility (unbound-to-bound iRMSD over 1.2 Å), ReplicaDock 2.0 successfully docks 61% of moderately flexible complexes and 35% of highly flexible complexes. Additionally, we demonstrate that by biasing backbone sampling particularly towards residues comprising flexible loops or hinge domains, highly flexible targets can be predicted to under 2 Å accuracy. This indicates that additional gains are possible when mobile protein segments are known.


EMS provider findings and interventions with a statewide EMS spine-assessment protocol.
To describe the utilization and findings with a statewide, prehospital spine-assessment protocol for emergency medical services (EMS) providers in a rural state. The study was a prospective sample of EMS patients evaluated by prehospital providers for trauma-related injury during a one-year investigation period. Prehospital providers prospectively completed supplementary spine data-collection forms that reported patient demographics and EMS provider findings with the spine-assessment protocol. Data were analyzed using descriptive statistics. There were 207,545 EMS encounters during the study period, including 31,885 transports for acute trauma-related illness. Prehospital providers provided spine-assessment forms for 2,220 patient encounters. Providers reported a decision to immobilize 1,301 (59%) patients. For these immobilized patients, spine protocol findings included 416 (32%) patients deemed as unreliable, 358 (28%) with distracting injury, 80 (6%) with an abnormal neurologic examination, and 709 (54%) with spine pain or tenderness. Linkage of EMS and hospital data revealed seven acute spine fracture patients among the 2,220 reported encounters, all of whom were immobilized by EMS providers. Use of this prehospital spine-assessment protocol resulted in an EMS provider decision not to immobilize approximately 40% of EMS trauma patients. Few spine fracture patients were encountered during the investigational period, though all were immobilized.


[Modifying the non-specific defense with bacteria, fungi and their metabolic products].
In a literature survey frequently mentioned bacterial and mycological modulators of the non-specific immune system are discussed. The substances are listed in a table pointing out indications respectively state of development. Commonly used definitions concerning substances that take influence in the non-specific immune system are discussed.


Challenges of Managing Multiple Myeloma Patients with Sickle Cell Disease: A Case Report and Review of Literature.
BACKGROUND A congenital hemolytic anemia, sickle cell disease can present with various clinical findings. Sickle cell disease is typically a disease of younger people and multiple myeloma typically occurs in older individuals. Multiple myeloma is rare among patients with sickle cell disease. Both multiple myeloma and sickle cell disease can cause various types of organ damage by different mechanisms. CASE REPORT We report a case of a patient who was born with sickle cell disease and presented with multiple myeloma later in life. Although he responded to anti-myeloma therapy, he died of hepatic and renal failure from complications of both multiple myeloma and sickle cell disease. CONCLUSIONS We discuss the complexity involved and present a review of the literature on managing multiple myeloma in relation to hepatic iron overload and end-stage renal disease in the setting of multiple myeloma and underlying sickle cell disease.


Clues and new evidences in arterial hypertension: unmasking the role of the chloride anion.
The present review will focus on the role of chloride anion in cardiovascular disease, with special emphasis in the development of hypertensive disease and vascular inflammation. It is known that acute and chronic overload of sodium chloride increase blood pressure and have pro-inflammatory and pro-fibrotic effects on different target organs, but it is unknown how chloride may influence these processes. Chloride anion is the predominant anion in the extracellular fluid and its intracellular concentration is dynamically regulated. As the queen of the electrolytes, it is of crucial importance to understand the physiological mechanisms that regulate the cellular handling of this anion including the different transporters and cellular chloride channels, which exert a variety of functions, such as regulation of cellular proliferation, differentiation, migration, apoptosis, intracellular pH and cellular redox state. In this article, we will also review the relationship between dietary, serum and intracellular chloride and how these different sources of chloride in the organism are affected in hypertension and their impact on cardiovascular disease. Additionally, we will discuss the approach of potential strategies that affect chloride handling and its potential effect on cardiovascular system, including pharmacological blockade of chloride channels and non-pharmacological interventions by replacing chloride by another anion.


Scleral buckling in 2 1/2 to 11-month-old premature infants with retinal detachment associated with acute retrolental fibroplasia.
The authors report their experience over the past 5 years in the cryosurgical management of retinal detachment in 10 eyes of 8 premature infants with retrolental fibroplasia, whose ages at the time of detachment surgery ranged from 2 1/2 months to 11 months, with an average of 5 1/2 months. Postoperative follow-up, ranging from 8 to 61 months, has revealed that scleral buckling was successful in 6 eyes (60%). The authors have found that ERG responses are slow to become established postoperatively, and have concluded that VER testing is of questionable value in these cases.


Insulin stimulates transport of organic anion compounds mediated by organic anion transporting polypeptide 2B1 in the human intestinal cell line Caco-2.
Organic anion transporting polypeptide 2B1 (OATP2B1) is the major uptake transporter in the intestine, and transports various clinically used therapeutic agents. Insulin acts through the insulin receptor in targeted cells, and Rab8A is one of the insulin signaling pathways. The small intestine in humans also expresses insulin receptor and Rab8A. It has been reported that insulin stimulates peptide transporter 1 (PEPT1) expression at the apical membrane and increases uptake of PEPT1 substrates in small intestine epithelial model cells (Caco-2 cells). However, the effect of insulin on OATP2B1 in the small intestine has not been fully investigated. We found that Rab8A was associated with OATP2B1-mediated estrone-3-sulfate (E3S) uptake. Insulin stimulated the uptake of E3S by Caco-2 cells and the enhancement was sustained for 120 min. The Vmax value of E3S uptake significantly increased upon insulin exposure. Caco-2 cells treated with insulin showed increased OATP2B1 expression at the cell surface. The apical-to-basal transport of E3S was also increased by insulin. The increase of E3S transport was inhibited by the cold condition (4 °C) or the OATP2B1 inhibitor, taurocholate. These results indicate that insulin acts on the small intestine to increase OATP2B1-mediated absorption.


Potential advantage of higher-order modes of birdcage coil for parallel imaging.
The potential advantages of the higher-order resonant modes of a low-pass birdcage coil for parallel imaging are investigated using FDTD simulations in a spherical phantom. Better parallel imaging performance can be achieved in axial planes than in sagittal planes. If more modes are employed, the average g-factor (gmean) and maximum g-factor (gmax) will be improved for a specific acceleration factor (R). G-factor performance at 3 T (gmean=1.79, gmax=3.55) and 7 T (gmean=1.60, gmax=2.45) can be achieved even with an acceleration factor as high as four when all six order modes of 12-rung low-pass birdcage coil are incorporated for imaging on a mid-axial plane. For a specific number of channels, the optimum combination of corresponding modes can be obtained for different acceleration factors. Based on the g-factor and SNR performance, the total degenerate multi-mode birdcage coil with six order resonant modes has better homogeneous coverage and SENSE performance than the 8-element phased array coil, although requiring fewer channels. In addition, the dielectric effects at high field can improve the parallel imaging performance.


Molecular mechanisms of action of glyburide on the beta cell.
A high-affinity sulfonylurea receptor has been identified on the plasma membrane of the beta cell. The potent second-generation sulfonylureas, glyburide and glipizide, saturate the receptor in the low nM concentration range, whereas first-generation drugs bind to and saturate the receptor in the microM range. For each of the sulfonylureas, there is excellent quantitative agreement among the equilibrium binding constant (Kd), the half-maximal inhibition of potassium ion (K+) efflux (K0.5), and the half-maximal stimulation of insulin secretion (ED50), when these values are obtained from insulin-secreting cell lines or from isolated mouse pancreatic islets. The inhibition of K+ efflux by the sulfonylureas, coupled with the sulfonylurea inhibition of the activity of a specific adenosine triphosphate (ATP)-sensitive K+ channel embedded in the plasma membrane of whole cells or in excised membrane patches, suggests that the sulfonylurea receptor is this channel protein or a closely associated subunit. The activity of the ATP-sensitive K+ channel is also controlled by the insulin secretagogues, glucose and certain amino acids. These compounds must be metabolized to inhibit the channel activity and appear to do so by increasing the level of ATP or by increasing the ATP/adenosine diphosphate (ADP) ratio. ATP reduces channel activity by binding to a specific nucleotide-binding site on the cytoplasmic surface of the protein. There is a synergy between the action of glucose and that of the sulfonylureas. The sulfonylureas, for example, are better effectors of insulin secretion in the presence of glucose. Inhibition of the ATP-sensitive K+ channels results in depolarization of the plasma membrane and a subsequent influx of extracellular calcium ions through voltage-dependent calcium channels. An increase in the free intracellular calcium level is the signal, or "second messenger," that triggers exocytosis and the release of insulin. The sulfonylurea receptor has a molecular weight of 140,000 and can be solubilized by digitonin, retaining the same rank order of sulfonylurea binding affinities as the membrane-bound protein. Several laboratories are currently purifying the receptor and/or cloning the receptor gene.


PET/MRI: Technical Challenges and Recent Advances.
Integrated positron emission tomography (PET)/magnetic resonance imaging (MRI), which can provide complementary functional and anatomical information about a specific organ or body system at the molecular level, has become a powerful imaging modality to understand the molecular biology details, disease mechanisms, and pharmacokinetics in animals and humans. Although the first experiment on the PET/MRI was performed in the early 1990s, its clinical application was accomplished in recent years because there were various technical challenges in integrating PET and MRI in a single system with minimum mutual interference between PET and MRI. This paper presents the technical challenges and recent advances in combining PET and MRI along with several approaches for improving PET image quality of the PET/MRI hybrid imaging system.


Comparative efficacy and safety of the minimally invasive ilioinguinal approach for anterior pelvic ring fracture: A protocol for systematic review and network meta-analysis.
Anterior pelvic ring contributes up to 40% of the stability of the pelvic ring and is located in close proximity to key pelvic organs, blood vessels, and nerves. An anterior pelvic ring fracture causes severe pain and is a potentially life-threatening condition in trauma patients. Currently available surgical repair methods are highly technical and include high risks of complications. The minimally invasive ilioinguinal approach (MIIA) is an emerging technique that reduces the risk of femoral nerve and external iliac vessel injury. However, the safety and efficacy of this technique have yet to be systematically scrutinized. This study outlines a proposed protocol for a network meta-analysis that investigates the efficacy of MIIA for anterior pelvic ring fracture. This study will utilize both Chinese and English language databases. All randomized controlled trials studying the use of MIIA for anterior pelvic ring fracture from January 2016 to May 2021 will be incorporated. Researchers will screen for literature that fits the inclusion criteria, followed by an assessment of risk bias and, finally, data extraction. The Bayesian network meta-analysis will be used to evaluate all available Stata 14.0 and WinBUGS software. Our research aims to uncover the clinical utility of the MIIA approach for anterior pelvic ring fractures. Not required. INPLASY2021110020.


In vitro profile of some opioid pentapeptide analogues.
The opiate agonist potency of thirteen synthetic enkephalin pentapeptides has been examined on the electrically stimulated guinea pig ileum and mouse vas deferens preparations in comparison with methionine and leucine enkephalins, beta-endorphin and normorphine. Their antagonism by naloxone (Ke) was also assessed on each preparation. Our findings are compatible with, and are discussed in the context of, the hypothesis that these preparations possess at least two populations of receptors.


Updated Campylobacter jejuni Capsule PCR Multiplex Typing System and Its Application to Clinical Isolates from South and Southeast Asia.
Campylobacter jejuni produces a polysaccharide capsule that is the major determinant of the Penner serotyping scheme. This passive slide agglutination typing system was developed in the early 1980's and was recognized for over two decades as the gold standard for C. jejuni typing. A preliminary multiplex PCR technique covering 17 serotypes was previously developed in order to replace this classic serotyping scheme. Here we report the completion of the multiplex PCR technology that is able to identify all the 47 Penner serotypes types known for C. jejuni. The number of capsule types represented within the 47 serotypes is 35. We have applied this method to a collection of 996 clinical isolates from Thailand, Cambodia and Nepal and were able to successfully determine capsule types of 98% of these.


Surgical management of primary liver cancers during the COVID-19 pandemic: overcoming the dilemma with standardization.
The present study evaluates the impact of the pandemic on outcomes after surgical treatment for primary liver cancer in a high-volume hepatopancreatobiliary surgery center. Patients, who underwent liver resection for primary liver resection between January 2019 and February 2020, comprised pre-pandemic control group. The pandemic period was divided into two timeframes: early pandemic (March 2020-January 2021) and late pandemic (February 2021-December 2021). Liver resections during 2022 were considered as the post-pandemic period. Peri-, and postoperative patient data were gathered from a prospectively maintained database. Two-hundred-eighty-one patients underwent liver resection for primary liver cancer. The number of procedures decreased by 37.1% during early phase of pandemic, but then increased by 66.7% during late phase, which was comparable to post-pandemic phase. Postoperative outcomes were similar between four phases. The duration of hospital stay was longer during the late phase, but not significantly different compared to other groups. Despite an initial reduction in number of surgeries, COVID-19 pandemic had no negative effect on outcomes of surgical treatment for primary liver cancer. The structured standard operating protocol in a high-volume and highly specialized surgical center can withstand negative effects, a pandemic may have on treatment of patients.


Bioluminescence imaging of hollow fibers in living animals: its application in monitoring molecular pathways.
We have applied noninvasive optical imaging technology to the in vivo hollow fiber assay, using tumor cell lines in which optical reporters are expressed in response to activation/inhibition of a specific molecular pathway. In vivo noninvasive imaging of molecular pathways in cells within hollow fibers enables a rapid and accurate evaluation of drug targets and provides useful insights to guide novel drug discovery. In this protocol we show, as an example, that a luciferase reporter, driven by the responsive element of nuclear factor NF-kappaB, was induced in cells within hollow fibers implanted in living mice, and a detailed procedure for in vivo bioluminescence imaging of hollow fibers is described. This approach can, in principle, be applied to image any molecular pathways of interest when appropriate reporter cells are generated. Hollow fiber encapsulation and implantation takes 2 d, and in vivo validation of reporter takes 1-2 weeks.


The management of musculoskeletal disorders in the workplace.
Musculoskeletal disorders are a major cause of suffering and disability among working-age adults. Although working in ergonomically unsound jobs may lead to the development of certain musculoskeletal disorders, it is increasingly recognised that well-designed work is generally good for health and individuals with musculoskeletal disorders generally benefit from working. This chapter explores how health-care professionals should assess patients' fitness for work, what factors should be considered and how the results should be communicated and to whom. Of necessity, this chapter describes current United Kingdom (UK) schemes and systems. Nevertheless, the principles described can be extended to most countries but the reader is advised to familiarise themselves with the detail of the equivalent national services in their own practice. The new UK Fit for Work service is explained together with advice on how best to use a fit note to optimise patients' short- and long-term health. We detail what benefits are available to those who are unable to work because of poor health and how health professionals can achieve an optimum balance between supporting those who are genuinely unfit to work through benefits from a welfare state and encouraging and facilitating those who can earn an independent living to do so.


Assessment of eye fatigue caused by head-mounted displays using eye-tracking.
Head-mounted displays (HMDs) and virtual reality (VR) have been frequently used in recent years, and a user's experience and computation efficiency could be assessed by mounting eye-trackers. However, in addition to visually induced motion sickness (VIMS), eye fatigue has increasingly emerged during and after the viewing experience, highlighting the necessity of quantitatively assessment of the detrimental effects. As no measurement method for the eye fatigue caused by HMDs has been widely accepted, we detected parameters related to optometry test. We proposed a novel computational approach for estimation of eye fatigue by providing various verifiable models. We implemented three classifications and two regressions to investigate different feature sets, which led to present two valid assessment models for eye fatigue by employing blinking features and eye movement features with the ground truth of indicators for optometry test. Three graded results and one continuous result were provided by each model, respectively, which caused the whole result to be repeatable and comparable. We showed differences between VIMS and eye fatigue, and we also presented a new scheme to assess eye fatigue of HMDs users by analysis of parameters of the eye tracker.


Bacterial Growth Control Mechanisms Inferred from Multivariate Statistical Analysis of Single-Cell Measurements.
Analysis of single-cell measurements of bacterial growth and division often relied on testing preconceived models of cell size control mechanisms. Such an approach could limit the scope of data analysis and prevent us from uncovering new information. Here, we take an "agnostic" approach by applying regression methods to multiple simultaneously measured cellular variables, which allow us to infer dependencies among those variables from their apparent correlations. Besides previously observed correlations attributed to particular cell size control mechanisms, we identify dependencies that point to potentially new mechanisms. In particular, cells born smaller than their sisters tend to grow faster and make up for the size difference acquired during division. We also find that sister cells are correlated beyond what single-cell, size-control models predict. These trends are consistently found in repeat experiments, although the dependencies vary quantitatively. Such variation highlights the sensitivity of cell growth to environmental variations and the limitation of currently used experimental setups.


Serotonin: good or bad for bone.
Besides its action as a neurotransmitter, serotonin has multiple physiological functions in several peripheral organs. Recently, Yadav et al. suggested that peripheral serotonin produced in the gut was a major negative regulator of osteoblast proliferation. These data were challenged by Cui et al. that showed no change in bone density in mature mice with a global invalidation of tryptophan hydroxylase 1, the enzyme responsible of serotonin synthesis in the periphery. In this context, we showed that osteoclasts are able to synthetize serotonin that acts locally to induce osteoclast precursors differentiation. Our data and previous results from others suggest that rather than acting as a hormone, serotonin produced in the bone could act locally on osteoclast and osteoblast realizing in the bone a complete micro-serotoninergic system.


Sham feeding of sucrose increases the ratio of 3,4-dihydroxyphenylacetic acid to dopamine in the hypothalamus.
Recent pharmacological experiments suggested that central dopaminergic (DA) mechanisms are necessary for the normal eating response to sweet stimuli. To test this hypothesis, we measured the ratio of dihydroxyphenylacetic acid (DOPAC) to dopamine (DA) in forebrain DA terminal fields during the sham feeding of sucrose (1-40%) by rats after 17 hr of food deprivation. After 9 min of sham feeding 10% or 40% sucrose, DOPAC/DA increased in the hypothalamus, but not in other forebrain regions including the n. accumbens, amygdala, and pyriform cortex. This increase in hypothalamic DOPAC/DA did not occur after 9 min of sham feeding 1%, 1.25%, or 2.5% sucrose. The increased DA metabolism required that sham feeding of 10% or 40% sucrose be maintained for longer than 3 min, because no increase of DOPAC/DA was observed in any forebrain region after 3 min of sham feeding. These results are strong evidence that hypothalamic DA mechanisms are activated by the sham feeding of sucrose solutions and they support the hypothesis that central DA mechanisms are necessary for the normal eating response to sweet stimuli.


MSX-1 gene expression and regulation in embryonic palatal tissue.
The palatal cleft seen in Msx-1 knock-out mice suggests a role for this gene in normal palate development. The cleft is presumed secondary to tooth and jaw malformations, since in situ hybridization suggests that Msx-1 mRNA is not highly expressed in developing palatal tissue. In this study we demonstrate, by Northern blot analysis, the expression of Msx-1, but not Msx-2, in the developing palate and in primary cultures of murine embryonic palate mesenchymal cells. Furthermore, we propose a role for Msx-1 in retinoic acid-induced cleft palate, since retinoic acid inhibits Msx-1 mRNA expression in palate mesenchymal cells. We also demonstrate that transforming growth factor beta inhibits Msx-1 mRNA expression in palate mesenchymal cells, with retinoic acid and transforming growth factor beta acting synergistically when added simultaneously to these cells. These data suggest a mechanistic interaction between retinoic acid, transforming growth factor beta, and Msx-1 in the etiology of retinoic acid-induced cleft palate.


Induction of an auto-anti-IgE response in rats I. Effects on serum IgE concentrations.
With a view to specifically suppressing the IgE isotype, rats of high (BN) and low (PVG.RT1u) IgE-responding phenotypes were immunized with a highly purified rat IgE myeloma (IR2) in an attempt to induce an anto-anti-IgE response. Rat IgE antibodies against epsilon determinants were detected in the serum of IR2-immunized animals using a solid-phase (plate) radioimmunoassay. The auto-anti-IgE antibodies detected were found to bind to IR2, to a second rat IgE myeloma (IR162) and to mouse monoclonal IgE but not rat IgG. The specificity of the anti-epsilon binding was shown by inhibition studies. The raising of an auto-anti-IgE response in PVG.RT1u rats severely depleted the serum level of circulating IgE for at least 8 weeks. In BN rats, immunization with IR2 caused marked fluctuations in serum IgE levels. The rats in both strains remained healthy throughout the experiment. The rate and route of IgE break down was not altered in anti-IgE-producing rats. The relevance of the present model in understanding and possibly controlling allergic disorders is considered.


The effect of direct acting antiviral agents on vascular endothelial function in Egyptian patients with chronic hepatitis C virus infection.
Chronic hepatitis C virus (HCV) infection is correlated with cerebrovascular and cardiovascular disease (CVD). This study aimed to assess the effect of treatment with DAAs on vascular endothelial function in cirrhotic and non-cirrhotic HCV infected patients without any CVD risk factors. Fifty chronic HCV genotype 4 infected patients, without cardiovascular risks who have been listed to receive sofosbuvir/daclatasvir with ribavirin combination as triple therapy for 3 months were prospectively recruited. Endothelial dysfunction markers as soluble vascular cell adhesion molecule-1 (sVCAM-1) and Von willebrand factor (vWf) and inflammation marker (IL6) were estimated at baseline and 3 months post the end of therapy (SVR). All patients achieved SVR. VCAM1 level was significantly improved after HCV clearance with DAA in cirrhotic HCV patients (P = 0.002) compared to patients with mild liver fibrosis (P = 0.006). Levels of vWF also decreased significantly in cirrhosis and non-cirrhosis groups after SVR (P < 0.001 and P = 0.011, respectively). Systemic inflammatory marker (IL6) showed significant decrease in cirrhotic patients (P = 0.001). While, IL6 level did not change significantly in non-cirrhotic group (P = 0.061). Also at SVR, noninvasive liver fibrosis indices have been reduced significantly in the two groups (P < 0.001). HCV clearance by new DAA treatment improves the vascular endothelial dysfunction in Egyptian HCV infected patients with different levels of liver fibrosis and with no risk factors for endothelial dysfunction or CVD.