Quantitative Image Analysis of Single-Molecule mRNA Dynamics in Living Cells.
Single mRNA molecules can be imaged in living cells by a method that consists in genetically inserting binding sites for a bacteriophage protein in the gene of interest. The resulting reporter transgene is then integrated in the genome of cells that express the phage protein fused to a fluorescent tag. Upon transcription, binding of the fluorescent protein to its target sequence makes the RNA visible. With this approach it is possible to track, in real time, the life cycle of a precursor mRNA at the site of transcription in the nucleus and transport of mature mRNA to the cytoplasm. In order to measure the fluorescence associated with individual RNA molecules over time, we developed a semi-automated quantitative image analysis tool termed STaQTool. We describe in detail the implementation and application of the STaQTool software package, which is a generic tool able to process large 4D datasets allowing quantitative studies of different steps in gene expression.


A deep-sea hydrogen peroxide-stable alkaline serine protease from Aspergillus flavus.
We report here the production of an alkaline serine protease by Aspergillus flavus isolated at 5600-m depth from deep-sea sediments of the Central Indian Basin. When grown on defatted groundnut oil meal at 30 °C for 48-72 h, this fungal isolate produced 2000-2500 ACU mL-1 of alkaline protease. The purified protease had activity optima at pH 10.0 and 45 °C. It was a thiol-independent serine protease, identified as an alkaline serine protease ALP1 with a molecular mass of 42.57 kDa. The thermostability and activity of the enzyme increased at 60 °C, in the presence of additives such as sucrose, Tween 20, sorbitol, Ca2+ and glycerol and was not adversely affected by H2O2 indicating its potential as a detergent additive.


Delayed appearance of wing-beating tremor after liver transplantation in a patient with Wilson disease.
Orthotopic liver transplantation (OLT) is the sole etiological treatment for Wilson disease (WD), but several neurological complications after OLT have been reported. We report a WD patient who developed a unilateral wing-beating tremor 6years after OLT. New neurological symptoms develop immediately after OLT in most cases. In our patient, the onset of extrapyramidal symptoms was at a prolonged interval after OLT. To our knowledge this is the first patient with delayed extrapyramidal symptoms after OLT in WD where the pathophysiology of these late extrapyramidal symptoms is still unknown.


Microanalysis of stomach cancer glycosaminoglycans.
Glycosaminoglycans (GAGS) are anionic, linear, polysaccharides involved in cell signaling. The GAG content, composition and structure of human tissue have been suggested to play a role in cancer and might provide useful diagnostic or prognostic markers. The current study examines 17 stomach tissue biopsy samples taken from normal individuals and from patients with gastric cancers. An ultrasensitive liquid chromatography (LC) - mass spectrometry assay was applied to individual biopsy samples as small 250 μg providing GAG content and disaccharide composition. The results of these analyses show a significant increase in non-sulfated chondroitin/dermatan sulfate concentration in all cancer samples when compared to normal tissues. In addition in advanced gastric cancer, a significant decrease is observed in hyaluronan.


The Role of Serum Calprotectin as a New Marker in Abdominal Aortic Aneurysms - A Preliminary Report.
Abdominal Aortic Aneurysm (AAA) remains a surgical challenge. There are many recognizable markers associated with the formation of AAA. Previous experiments carried out on animal models have shown a correlation between serum calprotectin and the occurrence of AAA. This study aimed to evaluate the level of calprotectin as a potential diagnostic biomarker in patients with diagnosed AAA. The study group consisted of 75 patients aged 35-75 years assigned to two groups: a control group (n=43) of healthy subjects without AAA and a study group (n=32) of patients with a diagnosed AAA. The first calprotectin test was performed upon patient admission to the hospital, and the second control test was performed after three months. The concentration of calprotectin in plasma was determined using the Immunoenzymatic Method (ELISA) with the commercially available Assaypro Kit (AssayMax™ Human Calprotectin ELISA Kit), as well as the sandwich method with polyclonal antibodies to human calprotectin and peroxidase enzyme. Serum calprotectin levels in AAA patients were three times higher than in healthy subjects (p<0.05). A statistically significant twofold decrease in calprotectin concentration was observed after AAA surgery compared to the control group (p<0.05). Calprotectin levels can be an important marker in the detection of AAA. In conclusion, AAA patients showed a threefold increase in serum calprotectin level and a twofold decrease in this marker after AAA surgery.


Shrinking the language accessibility gap: a mixed methods evaluation of telephone interpretation services in a large, diverse urban health care system.
Language interpretation services for patients who are not proficient in a country's official language(s) are essential for improving health equity across diverse populations, and achieving clinical safety and quality for both patients and providers. Nevertheless, overall use of these services remains low, regardless of how they are delivered. In Toronto, Ontario, one of the most ethnically diverse urban centres, the regional local health integration network which oversees the highest concentration of health care organizations servicing 1.2 million residents, partnered with key stakeholders to make Over-the-Phone (OPI) interpretation services broadly and economically available in 170 different languages to its diverse network of health care organizations. This evaluation aimed to assess patients' and providers' experiences with OPI in these varied settings and the impact (if any) on alternative interpretation services and on health service delivery access and quality. This study used a two-phased sequential exploratory mixed-methods approach to evaluate the initiative. Phase I was comprised of semi-structured interviews with representatives from the program stakeholders; these findings were applied to identify appropriate survey questions and response categories, and provided context and depth of understanding to Phase II results. Phase II included web-based and self-administered surveys for both providers and patients engaging with OPI. Both providers and patients identified a broad range of positive impacts OPI had on health care service delivery quality and access, and high levels of satisfaction with OPI, in a variety of health care settings. Providers also revealed a marked decrease in the use of ad-hoc, nonprofessional strategies for interpretation after the implementation of OPI, and noted it had either no impact on their workload or had decreased it overall. OPI is clearly not the sole answer to the complex array of health care needs and access gaps that exist for persons without proficiency in their country's official language. Nevertheless, this evaluation provides compelling evidence that OPI is a valuable component, and that it may contribute to a broader range of positive impacts, and within a broader range of health care settings, than previously explored.


Antimicrobial and phytochemical studies of Swertia corymbosa.
The aqueous, methanol, chloroform and hexane extracts of Swertia corymbosa were tested (in vitro) for their antimicrobial efficiency. Maximum inhibitory activity was noticed against Staphylococcus aureus and Salmonella typhi.


Nucleotide sequence of cloned cDNA for bovine corticotropin-beta-lipotropin precursor.
The nucleotide sequence of a 1,091-base pair cloned cDNA insert encoding bovine corticotropin-beta-lipotropin precursor mRNA is reported. The corresponding amino acid sequence indicates that the precursor protein consists of repetitive units and includes a third melanotropin sequence in its cryptic portion. Pairs of lysine and arginine residues separate the component peptides of the precursor.


Investigation of unilateral facial weakness: magnetic stimulation of the proximal facial nerve and of the face-associated motor cortex.
Twenty-four patients with unilateral facial weakness of various aetiologies were investigated using a magnetic stimulator to stimulate the proximal segment of the facial nerve directly (short latency response) and also to activate the facial motoneurons bilaterally via corticonuclear pathways by placing the stimulating coil over the motor cortex (long latency responses). Electromyographic recordings were taken from both mentalis muscles using concentric needle electrodes. Seventeen patients were investigated at various times after onset of idiopathic facial palsy (Bell's palsy). In the acute stage (less than 5 days after onset) short and long latency responses on the paretic side were abnormal, being absent in all but one patient, in whom the short latency response was delayed. These abnormal responses were the earliest neurographic correlate for nerve conduction block. In 4 out of 9 patients seen up to 30 days after onset of palsy, trans-synaptically evoked long latency responses were absent. In patients examined more than 2 months after onset, long latency responses could always be obtained and, in 5 of 8 patients, short latency responses could also be elicited, indicating a return of the direct excitability of the nerve. Five patients with cerebral hemisphere lesions causing mild unilateral facial weakness had absent long latency responses when stimulating over the affected hemisphere, but normal bilateral long latency responses following stimulation over the unaffected cerebral hemisphere; short latency responses were normal. Magnetic stimulation of the brain and of the facial nerve can differentiate between central and peripheral causes of unilateral facial weakness and may prove useful in the early assessment of the degree of conduction block in Bell's palsy.


Impact of Gender Differences on Outcomes of Peripheral Artery Disease Intervention (from a Nationwide Sample).
We aimed to evaluate the role of gender differences in the outcomes of catheter-based peripheral arterial disease interventions on a national level. We queried the National Inpatient Sample Database and identified all patients who presented with acute or symptomatic long term limb ischemia requiring transcatheter nonsurgical peripheral intervention in the years of 2016 to 2017. The primary outcome was major adverse cardiovascular events (MACE), defined as the composite end point of in-hospital mortality, nonfatal stroke, and acute myocardial infarction. Secondary outcomes were the subject components of the primary end point, vascular complications, major bleeding, acute kidney injury, limb amputation, total cost, and length of stay. A total of 58,165 patients were included. The majority were males (57.2%) and of white race (67.1%). On multivariate analysis, female gender was an independent predictor of MACE with an adjusted odd ratio (a-OR) of 1.36 (95% confidence interval [CI]: 1.12 to 1.65, p = 0.002), mortality (a-OR 1.52; 95% CI: 1.12 to 2.04, p = 0.006), nonfatal stroke (a-OR 2.51; 95% CI: 1.56 to 4.03, p < 0.001), major bleeding (a-OR 1.87; 95% CI: 1.53 to 2.28, p < 0.001), and higher cost with an adjusted mean ratio of 1.03 (95% CI: 1.00 to 1.06, p = 0.033). There was no significant difference in the rates of myocardial infarction, vascular complications, limb amputation, acute kidney injury, and length of stay. In conclusion, females presenting with acute or symptomatic long term limb ischemia requiring transcatheter peripheral intervention have a significantly higher composite risk of MACE.


Uneven damage on head and liner contact surfaces of a retrieved Co-Cr-based metal-on-metal hip joint bearing: An important reason for the high failure rate.
Detailed metallurgical investigations have been performed on a used Co-Cr-based metal-on-metal (MoM) hip joint bearing containing a type of liner that is commonly used in such joints. The damage on the metal-liner sliding surface was considerably more severe than that on the metal head counterpart, in terms of wear-scar density and width and microcrack frequency. Cross-sectional transmission electron microscopy revealed that a thick (>3 μm) nanocrystalline layer formed on the sliding surface of the head, whereas the liner had coarse carbides embedded in it and nanocrystals were formed in a very limited region no deeper than 1 μm. Comparative investigation of an unused head and a liner of identical type showed that although the chemical compositions of the liner and head were nearly identical, their microstructures were significantly different. Specifically, the grain size in the liner was larger than that in the head on average, and the grain boundaries of the liner were decorated with coarse carbides. Moreover, X-ray diffraction analysis revealed a large tensile residual stress only in the liner. These differences are possibly responsible for the wear damage on the liner being more serious than that on the head.


Feline oncornavirus-associated cell membrane antigen: a viral and not a cellularly coded transformation-specific antigen of cat lymphomas.
The feline oncornavirus-associated cell membrane antigen (FOCMA) was defined as a tumor antigen common to cat lymphomas and fibrosarcomas induced by feline leukemia virus (FeLV) and feline sarcoma virus (FeSV), respectively. The antigen was recognized by sera from cats thought to be resistant to leukemogenesis. We report here that a common denominator in the activity of naturally occurring viremic cat antisera to FOCMA is, in fact, their reactivity to FeLV C antigenic determinants. The cat antisera, monoclonal antibodies to FOCMA, and monoclonal antibodies to FeLV C, all reacted in immunofluorescence assays with FeLV C-infected cells and immunoprecipitated a molecule electrophoretically indistinguishable from envelope glycoprotein of FeLV. Viremic cat antisera to FOCMA bound to budding virus particles of FeLV C-infected cells, even though some of them could not be absorbed by mature virion proteins. Thus, the unusual feature of cat antibodies to FOCMA is their binding to nascent but not to mature virus particles. FOCMA-positive cat lymphomas expressed antigenic determinants of FeLV-C gp70, with or without productive infection. FeLV-negative tumors not expressing FeLV C gp70 were also FOCMA negative. Furthermore, most of the viremic cat sera and the monoclonal antibodies to FOCMA did not react with FeSV-transformed nonproducer cells. The absence of FOCMA from these cells and from FeLV-negative lymphoid tumors and its presence in FeLV-C infected fibroblasts indicated that this antigen is virus encoded and not a cellular tumor-specific antigen.


Sex-specific responses to GnRH challenge, but not food supply, in kittiwakes: Evidence for the "sensitivity to information" hypothesis.
Seasonal timing of breeding is usually considered to be triggered by endogenous responses linked to predictive cues (e.g., photoperiod) and supplementary cues that vary annually (e.g., food supply), but social cues are also important. Females may be more sensitive to supplementary cues because of their greater role in reproductive timing decisions, while males may only require predictive cues. We tested this hypothesis by food-supplementing female and male colonial seabirds (black-legged kittiwakes, Rissa tridactyla) during the pre-breeding season. We measured colony attendance via GPS devices, quantified pituitary and gonadal responses to gonadotropin releasing hormone (GnRH) challenge, and observed subsequent laying phenology. Food supplementation advanced laying phenology and increased colony attendance. While female pituitary responses to GnRH were consistent across the pre-breeding season, males showed a peak in pituitary sensitivity at approximately the same time that most females were initiating follicle development. The late peak in male pituitary response to GnRH questions a common assumption that males primarily rely on predictive cues (e.g., photoperiod) while females also rely on supplementary cues (e.g., food availability). Instead, male kittiwakes may integrate synchronising cues from their social environment to adjust their reproductive timing to coincide with female timing.


[Application of FTIR spectroscopy to the analysis of eleven kinds of Dendrobium].
To establish an FTIR method for the analysis of Dendrobium. Using fourier transform infrared spectrometer to record the characteristic spectra of eleven samples of Dendrobium, and to compare the spectra by PCA (principal component analysis). The FTIR spectra of the upper part of the stem displayed significant differences between fresh and dried samples of Dendrobium. On the other hand, differences were observed in the spectra of the middle and lower parts of stems of D. guangxieuse when compared to other species. The method of applying PCA to FTIR analysis is a rapid and dependable method for comparing samples of Dendrobium.


[Effects of the Saponins from Panax japonicus on Intestinal Epithelial Tight Junction Proteins in Aging Rats].
To investigate the moderating effects of total saponins of Panax japonicus on intestinal epithelial tight junction proteins in aging rats,and to explore the potential mechanism. SD rats were divided into adult group( 6 months),old model group( 24 months),and different doses( 10,30 and 60 mg / kg) of total saponins of Panax japonicus treatment groups. Levels of tight junction proteins( Occludin and ZO-1),anti-oxidative pathway proteins( Nrf2、HO-1 and NQO-1),mitochondrial biogenesis related proteins( Sirt1 and PGC-1α) and p-AMPK in the ileum were determined by immunohistochemistry staining. Compared with adult group,the expressions of Occludin,ZO-1,Nrf2,HO-1,NQO-1,Sirt1 and PGC-1α of aging rats were obviously decreased( P < 0. 01),and p-AMPK was inhibited in the ileum of aging rats. Compared with aging model rats,total saponins of Panax japonicus increased the expressions of Occludin,ZO-1,Nrf2,HO-1,NQO-1,Sirt1 and PGC-1α( P < 0. 05 or P < 0. 01),and activated p-AMPK in the ileum of aging rats. The decreased level of intestinal tight junction proteins in the ileum of aging rats may be related to oxidative stress. Total saponins of Panax japonicus can up-regulate the level of intestinal tight junction proteins to improve the intestinal mucosal barrier dysfunction in the ileum of aging rat via reducing the levels of oxidative stress.


Effects of Docetaxel Injection and Docetaxel Micelles on the Intestinal Barrier and Intestinal Microbiota.
Increasing evidence has suggested that chemotherapeutics affect the integrity of the intestinal barrier and alter the intestinal microbiota, thus limiting the therapeutic outcomes of cancer chemotherapy. Docetaxel (DTX) is used for breast cancer treatment and has gastrointestinal side effects, but the influence of DTX formulations on the intestinal barrier and intestinal microbiota remains unknown. Therefore, in this work, the influence of DTX injection (free DTX, commercial formulation) and DTX/methoxy poly(ethylene glycol)-block-poly(D,L-lactide) (mPEG-PDLLA) (DTX micelles, nanoformulation) on the integrity of the intestinal barrier and the intestinal microbiota is investigated. It is found that the free DTX causes significantly greater intestinal barrier damage than the DTX micelles. The diversity of the intestinal microbiota, and the relative abundance of Akkermansia muciniphila and Ruminococcus gnavus in the DTX micelle-treated group is significantly higher than that in the free DTX-treated group. Moreover, the tumor growth rate is elevated in antibiotic mixture-pretreated mice, demonstrating that the diversity and composition of the intestinal microbiota may be associated with tumor progression. This work demonstrates that different formulations of chemotherapeutics have different effects on the integrity of the intestinal barrier and the intestinal microbiota.


Sustainable Flow-Synthesis of (Bulky) Nucleoside Drugs by a Novel and Highly Stable Nucleoside Phosphorylase Immobilized on Reusable Supports.
The continuous synthesis of valuable nucleoside drugs was achieved in up to 99 % conversion by using a novel halotolerant purine nucleoside phosphorylase from Halomonas elongata (HePNP). HePNP showed an unprecedented tolerance to DMSO, usually required for substrate solubility, and could be immobilized on agarose microbeads through disulfide bonds, via a genetically fused Cystag. This covalent yet reversible binding chemistry showcased the reusability of agarose microbeads in a second round of enzyme immobilization with high reproducibility, reducing waste and increasing the sustainability of the process. Finally, the flow synthesis of a Nelarabine analogue (6-O-methyl guanosine) was optimized to full conversion on a 10 mm scale within 2 min residence time, obtaining the highest space-time yield (89 g L-1 h-1 ) reported to date. The cost-efficiency of the system was further enhanced by a catch-and-release strategy that allowed to recover and recirculate the excess of sugar donor from the downstream water waste.


Hypothesis:presynaptic receptors controlling renin release.
Presynaptic receptors have recently been identified on nerves supplying vascular smooth muscle. Renin is stored in juxtaglomerular cells in the wall of the afferent glomerular arteriole and the JG cell is probably derived from vascular smooth muscle. Nervous stimuli exert an important influence on release of renin, but the nature of the receptors involved is not agreed. We propose that there are presynaptic receptors associated with the JG apparatus as with vascular muscle and that a mechanism of this sort gives a better account of data on renin release.


Association of HLA class I alleles with aloplecia areata in Chinese Hans.
Some studies suggested that human HLA status may potentiate development of the AA phenotype and exists ethic differences. No report has been published about HLA class I alleles associated with AA in Chinese Hans. To study the distribution of HLA class I alleles and haplotypes in Chinese Hans AA patients and the relation of HLA class I profile with age of onset, severity, duration of current attack, past history and family history. The polymerase chain reaction-sequence-specific primer (PCR-SSP) method was used to analyze the distribution of HLA class I alleles in 192 patients with AA and 252 healthy controls in Chinese Hans. The frequencies of HLA-A*02, -A*03, -B*18, -B*27, -B*52 and -Cw*0704 were significantly higher in patients than in controls. The A*2-B*18, A*2-B*27, A*2-B*52, A*2-Cw*0704, B*18-Cw*0704, B*27-Cw*0704, B*52-Cw*0704 were found as high-risk haplotypes in developing AA in this study. The HLA-A*02 and -A*03 were observed increased frequencies in patients less than 50% hair loss, and HLA-B*27 equally in patients of 50-99% hair loss, alopecia totalis and alopecia universalis. The frequencies of HLA-A*02 and -B*27 were significantly raised in recurrent patients, and ones of HLA-A*02, -A*03 and -B*27 similarly in patients without a positive family history. This study demonstrated the positive association of HLA class I alleles and haplotypes with AA. There may be differences in genetic background in patients with different age of onset, grade of scalp hair loss, duration of current attack, a past history and a family history.


Latent membrane protein 1 of Epstein-Barr virus plays an important role in the serum starvation resistance of Epstein-Barr virus-immortalized B lymphocytes.
We have previously shown that SNU-1103, which is a latency type III Epstein-Barr virus (EBV)-transformed lymphoblastoid cell line (LCL) that was developed from a Korean cancer patient, resists serum starvation-induced G(1) arrest. In this study, we examined the role of latent membrane protein-1 (LMP-1) in serum starvation resistance, since LMP-1 is known to be essential for EBV-mediated immortalization of human B lymphocytes. The LMP-1 gene from SNU-1103 was introduced into the EBV-negative BJAB cell line, and shown to be associated with resistance to G(1) arrest during serum starvation. Western blot analyses of the LMP-1-transfected cells revealed several protein alterations as compared to vector-transfected control cells. The expression of key cell-cycle regulatory proteins was affected in the G(1) phase: the expression of cyclin D3, CDK2, p27, and E2F-4 was up-regulated, and the expression of cyclin D2, CDK6, p21, and p103 was down-regulated during serum starvation. These results imply that of the several EBV viral genes expressed in EBV-negative B lymphoma cells, LMP-1 mediates resistance to serum starvation-induced G(1) arrest. However, we cannot rule out the possibility that other EBV genes are also involved in the cell-cycle progression of the EBV-transformed LCL during serum starvation, since the altered protein expression profile of the LMP-1 transfectants was distinct from that of the SNU-1103 cells that expressed all of the EBV viral proteins.


Stereospecificity in allergic contact dermatitis to simple substituted methylene lactone derivatives.
The enantiomers of beta,gamma-dimethyl- and beta-methyl-alpha-methylene-gamma-butyrolactones have been synthesized stereospecifically from glutamic acid and beta-hydroxy isobutyric acid, respectively. Guinea pigs have been sensitized (Freund complete adjuvant technique) and tested to them. Both enantiomers of beta-methyl lactone as well as (+)-beta,gamma-dimethyl lactone induced enantiospecific allergic contact dermatitis (ACD); in turn, (-)-beta,gamma-dimethyl lactone showed no specificity. An interpretation is proposed.


Age and sex modulate metabolic and cardiovascular risk markers of patients after 1 year of highly active antiretroviral therapy (HAART).
Primary aim: To evaluate the modulating effects of age and sex on changes in plasma lipid levels in the response to highly active antiretroviral therapy (HAART). Secondary aim: To study insulin, leptin, adiponectin, E-selectin and P-selectin levels and their relation to demographics. Comparative, longitudinal, open cohort-study. Tertiary care center at a University Hospital. Eighty-two consecutive HIV-seropositive patients of either sex were enrolled in the study. Subjects were between 19 and 60 years of age and naive to HAART. Patients were treated with nucleoside reverse transcriptase inhibitors (NRTIs) and protease inhibitor(s) (PIs) or reverse transcriptase inhibitors (RTIs) only. Subjects were evaluated at baseline and after 3, 6 and 12 months. Total cholesterol levels increased in all patients. The greatest increase was seen in patients of older age treated with PI. Male gender was another risk factor for higher cholesterol but also for higher triglyceride levels. Therapy with protease inhibitors and/or stavudine had a negative influence on plasma triglyceride levels. Selectins, adipokines and insulin were less influenced by HAART. Based on the results of this study selection of therapy regimen according to the demographic risk factors sex and age can offer an easy strategy to help to minimize lipid elevations.


B-1 lymphocytes are able to produce IL-10, but is not pathogenic during Leishmania (Leishmania) amazonensis infection.
Over the years research has found an association between B lymphocytes and pathogenesis during Leishmania sp. infections. Recently we demonstrated that B-2 lymphocytes are the main producers of IL-10 during L. amazonensis infection, and that the disease severity in BALB/c mice was attributed to these IL-10-producing B-2 lymphocytes. Here, we aim to understand the role of peritoneal B-1 lymphocytes in the pathogenesis of L. amazonensis infection. We found that infection resulted in a decrease in the number of B-1a lymphocytes and increase in B-1b lymphocytes in the peritoneal cavity of WT BALB/c mice but not in B lymphocyte deficient mice (BALB/Xid) mice. In vitro interaction between B-1 lymphocytes and L. amazonensis showed that the amastigote form of the parasite was able to induce higher levels of IL-10 in B-1 lymphocytes derived from infected BALB/c mice than the promastigote. Moreover, B-1 lymphocytes derived from infected mice produced more IL-10 than B-1 lymphocytes derived from naïve mice under amastigote interaction. However, the repopulation of BALB/Xid mice with B-1 lymphocytes from WT BALB/c mice did not affect the lesion development. Together, these results suggest that although B-1 lymphocytes are able to produce IL-10 during in vitro interaction with L. amazonensis, they are not directly related to pathogenesis in vivo.


Glutamate current noise: post-synaptic channel kinetics investigated under voltage clamp.
1. Analysis of voltage-clamped noise has been used to investigate the operation of glutamate receptors and associated channels at the locust nerve-muscle junction. Channels opened by glutamate and an agonist have been compared. 2. Glutamate-induced current fluctuations have a power spectrum with a single (1/frequency2) component which fits a simple model for the operation of channels. The form of the spectra for glutamate voltage noise and for 'background' noise has been determined. 3. The single channel conductance was estimated from the spectra, gamma glutamate = 122 +/- 0.4 (S.E.) pS. This estimate is independent of membrane potential and of the amplitude of membrane current change produced by glutamate. 4. The rate constant, alpha, for the closing of glutamate-operated channels depends exponentially on membrane potential, conforming to the equation alpha = approximately alphaeetaVm (approximately alpha = 0.26 +/- 0.014 msec-1, eta = 0.0054 +/- 0.001 msec-1); the duration of the channel lifetime (tau) decreases with hyperpolarization. Membrane potential dependence of alpha reduces as temperature is lowered. 5. For glutamate-operated channels, the temperature dependence of alpha and gamma fits the Arrhenius equation; alpha and gamma decrease exponentially as a function of T-1 (degrees K) with a descrete change in slope at about 6 degrees C, indicating a change in the activaiton energies of the respective rate processes. 6. Spectra of quisqualate-induced current fluctuations have the same form as spectra for glutamate noise. The single channel conductance was estimated from the spectra, gamma quisqualate = 120 +/- 3.9 (S.E.) pS. 7. The rate constnt, alpha, for the closing of quisqualate-induced channels depends exponentially on membrane potential. The duration of the open state for quisqualate channels was 2.2 times longer than for glutamate channels. 8. For glutamate receptors the voltage-sensitivity of the channel life-time is in the opposite direction to that of ACh receptors in vertebrate muscle. Possible explanations for the sharp change in the activation energy of the rate processes associated with the channel are discussed.


Effect of cyclosporine on the renal tubular amino acid handling after kidney transplantation.
The renal tubular handling of free amino acids was studied 5-6 weeks after successful renal transplantation (tx) in 20 children treated with CsA and in 10 children treated with azathioprine (Aza). The results were compared with those of 34 control children. The amino-acid clearance studies were performed in combination with short-term inulin clearance. The CsA group revealed a mean inulin clearance of 49 +/- 16.8 ml/min/1.73 m2, the Aza group of 76.9 +/- 18.2, and the controls of 114 +/- 15.6. The plasma amino-acid concentrations were not different between CsA- and Aza-treated groups; however, most of the essential amino acids were lower in transplanted children than in controls. The decrease was correlated with the GFR. The amino-acid-clearance rates were statistically not different between both transplanted groups, but lower values than in controls were found for alanine, glycine, histidine, lysine, and phenylalanine, and significantly higher values for methionine. The fractional clearance rates of most amino acids were significantly elevated in transplanted children compared to controls. In CsA-treated patients, the fractional clearance rates of arginine, glycine, and serine were higher than in Aza-treated patients. No influence of CsA blood levels or rejection episodes on the amino-acid handling were detectable. We conclude that CsA has no specific influence on the renal handling of amino acids. Most disturbances observed depend on the graft function or may be caused by injuries to the graft following the tx procedure.


Missed lung cancer on chest radiography and computed tomography: imaging and medicolegal issues.
Missed lung cancer is an important medicolegal issue and is the second leading cause for malpractice actions against radiologists. Contributing factors to overlooked lung cancer can be ascribed to observer performance, lesion characteristics, and technical considerations. Of these, errors related to observer performance are probably the most important. Missed lung cancer does not necessarily constitute malpractice, but lesions of high conspicuity are more likely to be associated with an adverse legal outcome. Multiple strategies have been advocated to reduce the frequency of missed lung cancer. Several studies have emphasized the importance of careful comparison of the current radiograph with one or more prior examinations.


Should Non-ST-Elevation Myocardial Infarction be Treated like ST-Elevation Myocardial Infarction With Shorter Door-to-Balloon Time?
It is estimated that each year in the United States >780,000 persons will experience an acute coronary syndrome. Approximately 70% of these will have non-ST-elevation myocardial infarction (NSTEMI). Optimal timing of angiography in NSTEMI is a matter of debate. The aim of this retrospective analysis was to evaluate whether and how the timing of percutaneous coronary intervention (PCI) affects the 1-year rate of major adverse cardiac events (MACE) in patients presenting with NSTEMI. Within our PCI database, we identified 1550 patients who underwent PCI for NSTEMI. We then divided the population into 3 groups based on door-to-balloon time (D2BT) (group 1 = D2BT <90 minutes; group 2 = D2BT >90 minutes <24 hours; group 3 = D2BT >24 hours). Primary outcome was MACE, a composite of MI, death and target vessel revascularization (TVR), or TVR at 1 year. Baseline characteristics were heterogeneous among the 3 groups, with patients who underwent angiograms >24 hours from presentation being older with more cardiovascular co-morbidities. Patients with D2BT <90 minutes were more likely to present with cardiogenic shock and had higher troponin levels. In-hospital mortality was similar among the 3 groups, but 1-year MACE/TVR was significantly higher in groups 1 and 3, driven by worse mortality. In this large cohort of patients presenting with NSTEMI, patients who underwent PCI between 90 minutes to 24 hours from presentation had better 1-year outcomes but also had fewer co-morbidities and with significantly lower prevalence of cardiogenic shock and high troponin on presentation. Therefore, treatment selection bias makes causal inference concerning rapid revascularization and outcome unreliable. Randomized clinical trials are warranted to assess outcome of rapid revascularization in patients presenting with NSTEMI.


Defective vasodilation response to exercise in cutaneous precapillary vessels in diabetic humans.
Although anatomic abnormalities in capillary structure are well described in diabetes mellitus, physiologic responses of diabetic microcirculation are less clear. This study addresses whether cutaneous vasodilation occurs normally in diabetic subjects in response to a standard exercise of underlying muscle. The methods involve a washout analysis from forearm skin of the fast-moving isotope 133Xe and the diffusion-limited marker 111In-diethylenetriamine pentaacetic acid. Twenty-four normotensive type II diabetic patients of age (mean +/- SE) 59 +/- 1.4 yr were age and weight matched with 24 controls. Control subjects increased flow 50.6 +/- 6.4%, which was not different from the increase in diabetic subjects of 52.7 +/- 7.6%. Permeability-surface-area product (PS) increased in control subjects from 1.77 +/- 0.20 ml.min-1.100 g-1 to 3.79 +/- 0.50 (2P = .0001), whereas diabetic subjects did not show a change in PS (3.29 +/- 0.35 to 2.69 +/- 0.31). Resting PS was higher in diabetic than control subjects (P = .0003), perhaps indicating an already recruited capillary bed. During repetitive exercise, control subjects increased capillary density 125 +/- 27% (2P = .0001) above basal with a nonsignificant 14 +/- 8.4% decrease in mean capillary blood flow. Diabetic subjects responded with a nonsignificant decrease in capillary density and a significant 139 +/- 38% increase in capillary blood flow (2P = .001). It is concluded that, although overall vasodilation occurs normally in diabetic cutaneous circulation, the mechanism is different from the normal response in that flow increases by augmentation of capillary flow rather than by recruitment. This is consistent with the thesis promulgated by others that diabetic precapillary vessels have depressed vasodilatory responses or are already maximally vasodilated. The ability of diabetic subjects to vasodilate by augmentation was not correlated with years of diabetes or use of insulin and only slightly negatively correlated with random blood glucose levels (r = -.423, P less than .05).


Efficacy and implementation of an Internet psychoeducational program for teens with type 1 diabetes.
The purpose of the study was to evaluate the participation and preliminary efficacy of an Internet psychoeducational program (Teens.Connect) shown to be efficacious under controlled conditions compared with an open-access diabetes website for youth (Planet D) on the primary outcomes of A1C and quality of life (QoL), and secondary outcomes of psychosocial and behavioral factors. Teens with type 1 diabetes (n = 124, 11-14 yr) from two clinical sites were randomly prescribed one of the programs and completed baseline, 3-month and 6-month data. A1C was obtained from clinic records. Participation data included number of log ins, posts to the discussion board, and lessons completed (Teens.Connect only). Descriptive and mixed model analyses were used. Eighty-five percent (85%) of consented teens registered for their prescribed program. Satisfaction and log ins were similar between groups (satisfaction ranged 3.3-3.5/5; mean log ins = 14/teen). Posts to the discussion forum were higher in Planet D (mean = 28 vs. 19). Participation in the Teens.Connect lessons was low, with only 69% of teens completing any lesson. After 6 months there were no significant differences in A1C, QoL or secondary outcomes between groups. Teens in the Teens.Connect group reported lower perceived stress over time (p < 0.01). Teens do not actively participate in an Internet psychoeducational program when they do not have frequent reminders, which may have contributed to a lack of treatment effect. Teens have many competing demands. Strategic implementation that includes targeted reminders and family support may be necessary to assure participation and improvement in health outcomes.


Unlocking new alleles for leaf rust resistance in the Vavilov wheat collection.
Thirteen potentially new leaf rust resistance loci were identified in a Vavilov wheat diversity panel. We demonstrated the potential of allele stacking to strengthen resistance against this important pathogen. Leaf rust (LR) caused by Puccinia triticina is an important disease of wheat (Triticum aestivum L.), and the deployment of genetically resistant cultivars is the most viable strategy to minimise yield losses. In this study, we evaluated a diversity panel of 295 bread wheat accessions from the N. I. Vavilov Institute of Plant Genetic Resources (St Petersburg, Russia) for LR resistance and performed genome-wide association studies (GWAS) using 10,748 polymorphic DArT-seq markers. The diversity panel was evaluated at seedling and adult plant growth stages using three P. triticina pathotypes prevalent in Australia. GWAS was applied to 11 phenotypic data sets which identified a total of 52 significant marker-trait associations representing 31 quantitative trait loci (QTL). Among them, 29 QTL were associated with adult plant resistance (APR). Of the 31 QTL, 13 were considered potentially new loci, whereas 4 co-located with previously catalogued Lr genes and 14 aligned to regions reported in other GWAS and genomic prediction studies. One seedling LR resistance QTL located on chromosome 3A showed pronounced levels of linkage disequilibrium among markers (r 2 = 0.7), suggested a high allelic fixation. Subsequent haplotype analysis for this region found seven haplotype variants, of which two were strongly associated with LR resistance at seedling stage. Similarly, analysis of an APR QTL on chromosome 7B revealed 22 variants, of which 4 were associated with resistance at the adult plant stage. Furthermore, most of the tested lines in the diversity panel carried 10 or more combined resistance-associated marker alleles, highlighting the potential of allele stacking for long-lasting resistance.


Best Predictors of Continuous Positive Airway Pressure Adherence.
About half of continuous positive airway pressure (CPAP)-treated patients are adherent, substantially affecting efficacy. A limitation to understanding predictors of adherence is the lack of a singular definition. Univariate analyses have suggested an array of factors that are statistically significant and reflect disease pathophysiology, clinical features, demographic characteristics, device-related variables, and psychological factors, but whether differences are clinically meaningful is unclear. There have been limited applications of multiple regression to compare the relative influence of multiple variables. This review article considers categories of variables that have been explored and suggests those that may be labeled "best" predictors in understanding CPAP use.


A comparison of depth of tissue injury caused by different modes of electrosurgical current in a pig colon model.
This study compared tissue injury produced by 3 types of electrosurgical current (pure cut, coagulation, blend) and two different procedures (snare and "hot biopsy"). Each type of electrosurgical current was passed through the colon of a live pig by means of a snare or "hot biopsy" forceps, and the depth of tissue injury was determined histologically. The high-frequency electrosurgical current was provided by a single electrosurgical generator. The peak power for each type of current was determined with an oscilloscope. The depth of tissue injury caused by coagulation current was significantly greater compared with blended current (p=0.0157). The depth of injury with coagulation current also was greater than with pure cut current, (p=0.0461 in a single statistical test; significance removed by Bonferroni-Dunn correction). With the hot biopsy forceps, the depth of tissue injury was deeper compared with that produced with a snare, regardless of the diameter of the snare loop. Peak power at a setting of 30 W was 1154 W for coagulation, 90.2 W for pure cut, and 227.8 W for blend current. When a high-frequency electrosurgical current device is used, use of a cutting current in the blend mode is recommended instead of coagulation current because this waveform is suitable for incision and provides effective hemostasis. Skillful technique is required for safe use of a hot biopsy forceps, because there is a significant potential for deeper tissue injury with this device.


Co-expression of protein phosphatases in insect cells affects phosphorylation status and expression levels of proteins.
The activity of kinases is regulated by phosphorylation on Ser, Thr or Tyr residues within the activation loop. The ability to produce these enzymes recombinantly with a specific phosphorylation status is essential in order to understand structure and function. In this paper we describe a screening approach to co-express different phosphatases together with a kinase in the baculovirus expression system. This enabled the testing of different phosphatases as well as different levels of both phosphatase and kinase by varying the multiplicity of infection (MOI) of the different baculoviruses. This approach translated well to a larger scale. An unexpected observation was that co-expression of the phosphatase could have profound effects on expression levels even of heterologous target proteins that would not be a substrate for the phosphatase. This was most apparent with lambda phosphatase, an enzyme that removes phosphorylation from Ser and Thr residues, where expression was almost completely abolished for all proteins, even at modest MOIs. The effect of lambda phosphatase was observed irrespective of whether co-expression was from two separate baculoviruses or from two genes on the same vector. The effect was shown to be due, in part at least, to a decrease in transcription.


Optical monitoring of activity of many neurons in invertebrate ganglia during behaviors.
Optical methods for monitoring neuron activity were developed because these methods lend themselves to simultaneous multiple-site measurements. With the use of new voltage-sensitive dyes, the dye-related pharmacology and photodynamic damage appear to be relatively unimportant. Using multiple-site measurements made with a 124-element photodiode array, we estimated that approximately 30 of the 200 neurons present in the Navanax buccal ganglion make action potentials during feeding and that approximately 300 of the 1100 neurons present in the Aplysia abdominal ganglion are active during the gill-withdrawal reflex. The fact that a light mechanical touch to the siphon skin activated such a large number of neurons in the abdominal ganglion suggests that understanding the neuronal basis of the gill-withdrawal reflex and its behavioral plasticity may be forbiddingly difficult.


Disrupting coupling within mycobacterial F-ATP synthases subunit ε causes dysregulated energy production and cell wall biosynthesis.
The dynamic interaction of the N- and C-terminal domains of mycobacterial F-ATP synthase subunit ε is proposed to contribute to efficient coupling of H+-translocation and ATP synthesis. Here, we investigate crosstalk between both subunit ε domains by introducing chromosomal atpC missense mutations in the C-terminal helix 2 of ε predicted to disrupt inter domain and subunit ε-α crosstalk and therefore coupling. The ε mutant εR105A,R111A,R113A,R115A (ε4A) showed decreased intracellular ATP, slower growth rates and lower molar growth yields on non-fermentable carbon sources. Cellular respiration and metabolism were all accelerated in the mutant strain indicative of dysregulated oxidative phosphorylation. The ε4A mutant exhibited an altered colony morphology and was hypersusceptible to cell wall-acting antimicrobials suggesting defective cell wall biosynthesis. In silico screening identified a novel mycobacterial F-ATP synthase inhibitor disrupting ε's coupling activity demonstrating the potential to advance this regulation as a new area for mycobacterial F-ATP synthase inhibitor development.


PAF-stimulated protein tyrosine phosphorylation in hippocampus: involvement of NO synthase.
The effect of platelet-activating factor (PAF) on protein tyrosine phosphorylation was studied in rat hippocampal slices. PAF caused an increase in the tyrosine phosphorylation of two phosphoproteins, which we identified by immunoprecipitation assays as the focal adhesion kinase p125FAK and crk-associated substrate p130Ca. The PAF effect was time- and dose-dependent. In addition, the involvement of PAF receptor was demonstrated by using PCA-4248, a specific receptor antagonist. When NO synthase was inhibited by NG-monomethyl-L-arginine (L-NMA), PAF-stimulated protein tyrosine phosphorylation was inhibited. In conclusion, our results indicate that PAF increased the tyrosine phosphorylation of both p125FAK and p130Cas proteins by the production of NO in hippocampus, suggesting that PAF may play a role in the functioning of this cerebral area.


Bilateral pneumothorax secondary to metastatic angiosarcoma of the breast.
Bilateral spontaneous pneumothorax is a rare complication of pulmonary metastases. We report a 44 year old woman who developed this complication a few months after undergoing mastectomy and postoperative radiotherapy for primary angiosarcoma of the breast.


Gene delivery into neuronal and glial cells by using a replication-deficient adenovirus vector: prospects for neurological gene therapy.
We have used a recombinant adenovirus vector (E1-) expressing β-galactosidase to explore a novel mechanism with which to transfer genes into cells of the central nervous system (CNS). The replication-deficient adenovirus vector expressing β-galactosidase (RAd35) was propagated on a permissive helper cell line (293 cells). High level protein expression from the human cytomegalovirus immediate early promoter (hCMV IE) was obtained in a target cell population of RAd35 infected cultured neuronal and glial cell lines. Light microscopy showed that over 50% of the glial cells studied expressed β-galactosidase. Following retinoic acid treatment, RAd35 infected cell lines ND7/23, NG108 and NTera2, showed β-galactosidase expression in up to 90% of the cells. In addition, these cells showed morphological evidence of differentiation into neurons. This pattern of β-galactosidase expression was also observed in primary rat cerebella granule neuron cultures. In vivo studies were performed in Balb/c mice following direct intracranial injections of RAd35 into the brain. Cell sections showed a localised staining in the brain at the site of injection of the virus. Non-replicating adenovirus vectors are therefore highly efficient systems for delivering a transgene into brain cells. However, their broad cell tropism may limit their applications for genetic disorders in which a specific cell type is to be targeted for gene therapy. To address this problem, we have constructed adenovirus vectors which contain specific neuronal promoters and are currently assessing in vitro expression.


Heterogenous electro-Fenton degradation of sulfamethoxazole on a polyethylene glycol-coated magnetite nanoparticles catalyst.
We report the preparation and application of poly (ethylene) glycol (PEG) coated magnetite nanoparticles (MNPs) catalyst for the heterogeneous electro-Fenton (HEF) degradation of sulfamethoxazole in real wastewater PEG-coated MNPs of four MNP:PEG ratios were synthesised using the co-precipitation method. The synthesised MNP were characterised using FTIR, XRD, EDX, TEM, and CHN elemental analysis. It was observed that the coating of MNP with PEG influences the nanoparticle size, agglomeration tendencies and catalytic efficiency of MNPs properties in the HEF degradation process. A 1:1 optimal MNP:PEG catalyst yielded 91% sulfamethoxazole degradation and 48% total organic carbon removal in 60 min, which is an improvement of 11% over degradation with the uncoated MNP. The PEG-coated MNP showed higher stability in 10 consecutive reaction cycles, reduced leaching, and improved performance at a lower dosage and broader pH range than the uncoated MNPs. These results show that coating MNP with PEG enhances HEF catalytic performance in the degradation of sulfamethoxazole in wastewater.


Characterizing long-COVID brain fog: a retrospective cohort study.
Long COVID or post-COVID condition (PCC) is a common complication following acute COVID-19 infection. PCC is a multi-systems disease with neurocognitive impairment frequently reported regardless of age. Little is known about the risk factors, associated biomarkers and clinical trajectory of patients with this symptom. To determine differences in clinical risk factors, associated biochemical markers and longitudinal clinical trajectories between patients with PCC with subjective neurocognitive symptoms (NC+) or without (NC-). A retrospective longitudinal cohort study was performed using a well-characterized provincial database of patients with clinically confirmed PCC separated into NC+ and NC- cohorts. Demographical, clinical and biochemical differences at initial consultation between the two patient cohorts were analyzed in cross-section. Multivariate regression analyses were conducted to identify independent risk factors for neurocognitive impairment. Determination of the recovery trajectory was performed using serial assessments of the patient-reported health-related quality of life (HR-QoL) metric Eq-5D-5L-vas score. Women, milder acute infection and pre-existing mental health diagnoses were independently associated with subjective neurocognitive impairment at 8 months post-infection. NC + patients demonstrated lower levels of IgG, IgG1 and IgG3 compared to NC- patients. The NC + cohort had poorer HR-QoL at initial consultation 8 months post-infection with gradual improvement over 20 months post-infection. Neurocognitive impairment represents a severe phenotype of PCC, associated with unique risk factors, aberrancy in immune response and a delayed recovery trajectory. Those with risk factors for neurocognitive impairment can be identified early in the disease trajectory for more intense medical follow-up.


Effect of sample matrices on supported liquid membrane: Efficient electromembrane extraction of cathinones from biological samples.
In this work, the effect of sample matrix on electromembrane extraction (EME) was investigated for the first time using cathinones (log P < 1.0) as polar basic model analytes. Ten supported liquid membranes (SLMs) were tested for EME from spiked buffer solutions, urine, and whole blood samples, respectively. For buffer solutions, SLMs containing aromatic solvents provided higher EME recovery than non-aromatic solvents, which confirmed the significance of cation-π interactions for EME of basic substances. Interestingly, when applied to urine and whole blood samples, aromatic SLMs were less efficient, while non-aromatic SLMs containing abundant hydrogen-bond acidity/basicity were efficient. These observations were explained by SLM fouling, and the antifouling property of the SLM was clearly dependent on the nature of the SLM solvent. Accordingly, a binary SLM containing aromatic 1-ethyl-2-nitrobenzene (ENB) and non-aromatic 1-undecanol (1:1 v/v) was developed. This binary SLM was not prone to fouling, and provided high recoveries of cathinones from urine and whole blood. EME based on this SLM was optimized and evaluated in combination with liquid chromatography tandem mass spectrometry (LC-MS/MS), and the linear ranges with R2 ≥ 0.9903 for cathinones in whole blood and urine were 5-200 ng/mL and 1-200 ng/mL, respectively. The LOD and LOQ of cathinones were ranged from 0.12 to 0.54 ng/mL and 0.38-1.78 ng/mL, respectively. The repeatability and accuracy bias at three levels were ≤11% and within 10%, respectively. In addition, the matrix effect ranged from 88% to 118% was also in compliance with guidelines for bioanalytical method validation provided by the European Medicines Agency.


Hamstring Muscle Quality Properties Using Texture Analysis of Ultrasound Images.
The aim of this study was to examine the intra- and inter-muscular differences of the hamstring muscles using textural analysis of ultrasound (US) images, and the relationship between textural indicators with hamstring torque. Transverse US scans were obtained from 10 young males from four different measurement sites along the thigh of each individual hamstring muscle at rest. Maximum-knee-flexion isometric torque measurements were also obtained. Texture analysis was applied to US images, and five gray-level co-occurrence matrix (GLCM) features were quantified: entropy (ENT), angular second moment (ASM), inverse difference moment (IDM), contrast (CON) and correlation (COR). The intraclass correlation coefficients ranged from 0.77 to 0.99, and the standard error of measurement ranged from 0.06 to 10.05%, indicating high test-retest reliability. Analysis of the variance indicated significant differences between measurement sites and individual muscles, with the proximal measurement sites having greater values for ASM, IDM and COR and lower values for ENT and CON compared with the distal sites. Additionally, only the COR at the proximal measurement site exhibited a significant relationship (r = -0.66) with strength. The present study indicated significant differences among hamstrings and measurement locations with respect to the textural analysis and may provide a novel indicator of hamstring functional properties.


Car drivers' attitudes towards motorcyclists: a survey.
Motorcyclists are over-represented in UK traffic accident statistics. Many car-motorcycle accidents are however due to the inappropriate actions of car drivers. It is predicted that car drivers at risk of collision with motorcycles have divergent attitudes and beliefs about motorcyclists compared to safer drivers, which may lead to a deficient mental model guiding their interactions with motorcyclists. To assess car drivers' attitudes towards motorcyclists, a survey was undertaken. Respondents filled in 26 general and motorcycle-related items and the 24 items of the reduced Driver Behaviour Questionnaire. Compared to an experienced dual driver group (who both drive cars and ride motorcycles), all other drivers showed divergent beliefs and attitudes. Four factors were extracted from the motorcycle items: negative attitudes, empathic attitudes, awareness of perceptual problems, and spatial understanding. Car drivers with a moderate amount of experience (between 2 and 10 years driving) held the most negative views and reported the most violations. The results have lead to several suggestions for interventions aimed at decreasing the divergence between drivers' perceptions of motorcyclists, and the perceptions of experienced dual drivers.


NADPH oxidase derived reactive oxygen species are involved in human neutrophil IL-1β secretion but not in inflammasome activation.
Neutrophils are essential players in acute inflammatory responses. Upon stimulation, neutrophils activate NADPH oxidase, generating an array of reactive oxygen species (ROS). Interleukin-1 beta (IL-1β) is a major proinflammatory cytokine synthesized as a precursor that has to be proteolytically processed to become biologically active. The role of ROS in IL-1β processing is still controversial and has not been previously studied in neutrophils. We report here that IL-1β processing in human neutrophils is dependent on caspase-1 and on the serine proteases elastase and/or proteinase 3. NADPH oxidase deficient neutrophils activated caspase-1 and did not exhibit differences in NALP3 expression, indicating that ROS are neither required for inflammasome activation nor for its priming, as has been reported for macrophages. Strikingly, ROS exerted opposite effects on the processing and secretion of IL-1β; whereas ROS negatively controlled caspase-1 activity, as reported in mononuclear phagocytes, ROS were found to be necessary for the exportation of mature IL-1β out of the cell, a role never previously described. The complex ROS-mediated regulation of neutrophil IL-1β secretion might constitute a physiological mechanism to control IL-1β-dependent inflammatory processes where neutrophils play a crucial role.


Nasal meningoencephalocele: A retrospective study of clinicopathological features and diagnosis of 16 patients.
Nasal meningoencephalocele (encephalocele or cephalocele) is a rare condition with congenital, traumatic, or spontaneous origins. We investigated the clinicopathological characteristics of nasal encephaloceles to improve pathologists' and clinicians' understanding of this disease. Sixteen patients with nasal encephaloceles were enrolled in this retrospective study investigating the condition's clinical and morphological features. Patients' average age was 37.8 (±20.8) years. The ratio of men to women was 2.2:1, patients' mean age was 47.4 (±11.8) years, and 10/16 patients had spontaneous encephaloceles. All patients with traumatic and spontaneous encephaloceles presented with cerebrospinal fluid leak. In 9/16 patients, the skull defect site occurred on the lateral wall of the sphenoid sinus. Both congenital patients experienced nasal obstruction. Histopathology, herniated tissues were brain and/or meningeal tissue, and the brain tissue was almost mature glial tissue. Nasal meningoencephalocele is a rare condition that can be challenging to diagnose. In patients with recurrent clear nasal discharge or in children with a unilateral nasal mass, a high index of suspicion for encephalocele is essential. In this study, spontaneous cases were most common in adults, and the lateral wall of the sphenoid sinus was the most common location.


An American Association for the Surgery of Trauma (AAST) prospective multi-center research protocol: outcomes of urethral realignment versus suprapubic cystostomy after pelvic fracture urethral injury.
Pelvic fracture urethral injuries (PFUI) occur in up to 10% of pelvic fractures. It remains controversial whether initial primary urethral realignment (PR) after PFUI decreases the incidence of urethral obstruction and the need for subsequent urethral procedures. We present methodology for a prospective cohort study analyzing the outcomes of PR versus suprapubic cystostomy tube (SPT) after PFUI. A prospective cohort trial was designed to compare outcomes between PR (group 1) and SPT placement (group 2). Centers are assigned to a group upon entry into the study. All patients will undergo retrograde attempted catheter placement; if this fails a cystoscopy exam is done to confirm a complete urethral disruption and attempt at gentle retrograde catheter placement. If catheter placement fails, group 1 will undergo urethral realignment and group 2 will undergo SPT. The primary outcome measure will be the rate of urethral obstruction preventing atraumatic passage of a flexible cystoscope. Secondary outcome measures include: subsequent urethral interventions, post-injury complications, urethroplasty complexity, erectile dysfunction (ED) and urinary incontinence rates. Prior studies demonstrate PR is associated with a 15% to 50% reduction in urethral obstruction. Ninety-six men (48 per treatment group) are required to detect a 15% treatment effect (80% power, 0.05 significance level, 20% loss to follow up/death rate). Busy trauma centers treat complete PFUI approximately 1-6 times per year, thus our goal is to recruit 25 trauma centers and enroll patients for 3 years with a goal of 100 or more total patients with complete urethral disruption. The proposed prospective multi-institutional cohort study should determine the utility of acute urethral realignment after PFUI.


Two-photon excited fluorescent silica nanoparticles loaded with iron(II) as a probe for determination and imaging of hydrogen peroxide in living cells.
A method is described for determination and optical imaging of hydrogen peroxide (H2O2) by using the two-photon (TP) excited fluorescence of silica (SiO2) nanoparticles containing Fe(II) ions. In the presence of H2O2, hydroxyl radicals (•OH) are produced via the Fenton reaction. This leads to quenching of the green fluorescence of a TP-excitable organic dye loaded into the SiO2NPs. Fluorescence is excited at 370 nm and has an emission peaking at 447 nm. The degree of quenching increases linearly in the 2.5 to 100 μM H2O2 concentration range. The nanoprobe is highly selective and sensitive, with a detection limit of 336 nM. The nanoprobe is biocompatible and was successfully used to image changes in the H2O2 concentration in HeLa cells via TP fluorescence imaging. Graphical abstractSchematic rpresentation of the detection of H2O2 by using the two-photon excited fluorescence of silica nanoparticles (TP-SiO2NPs) containing Fe2+. H2O2 triggers the Fenton reaction to produce hydroxyl radicals (•OH), which quench the green fluorescence of the SiO2NPs.


Biodegradable open cell foams of telechelic poly(epsilon-caprolactone) macroligand with ruthenium (II) chromophoric subunits via sub-critical CO2 processing.
This work reports on the effect of CO2 at subcritical conditions and the gaseous state on the telechelic poly(epsilon-caprolactone) polymers. The tested polymers are semi-crystalline in nature and thus the effect of functional groups and their overall contribution to foaming and formation of microstructures with open-cell morphollogy is discussed.


Attentional capture by auto- and allo-cues.
In a host of studies, the ability of various types of cues to capture attention has been examined. This article reviews a number of these studies by organizing them into a classification scheme based on the relationship between the putative attention-capturing item (the cue) and the item used to assess the distribution of attention (the probe). The second dimension of this taxonomy divides paradigms of attentional capture into those in which capture is indexed by performance benefits and those in which capture is indexed by performance costs. The relative methodological merits and disadvantages of the paradigms that occupy each of the cells of the resulting two-by-two matrix are discussed. A final section offers a new interpretation of the finding that dynamic cues capture attention.


Acute eosinophilic pneumonia in twins.
The pathogenesis of eosinophilic pneumonia is currently poorly understood, and this disease has not been reported in twins since 1983. Herein, we report a case of acute eosinophilic pneumonia in twins, which appeared to be triggered by initial smoking at different times by both patients. One patient resumed smoking after recovering from eosinophilic pneumonia, with no observed recurrence. This study discussed the possibility of an association between susceptibility to eosinophilic pneumonia and genetic factors in twins.


[Decline of competence in aging: an evaluation of the literature on "activities of daily living" and need for nursing care].
The results of a quantitative and qualitative analysis of 36 epidemiological studies on competence in activities of daily living and basic care needs in old age is presented. On the level of basic activities (ADL), the mean portions of elderly people in private households receiving help varied between 1.4% (eating) and 7.7% (bathing). With respect to the "instrumental" activities (IADL) cooking and shopping, the mean percentages were 14.1% and 17.8%, respectively. The mean rate of care needs in the elderly at home amounted to 4.7%. These global scores are further differentiated with respect to person variables (age, sex) and study characteristics (severity of care needs, inclusion of institutionalized elderly).


Cells transformed by a wide variety of agents express higher abundance levels of some cellular RNA species.
A cDNA-cloned library was prepared from mRNA synthesized by SV40-transformed mouse cells. Eleven cDNA clones were selected based on their ability to hybridize higher levels of mRNA in SV40-transformed 3T3 cells than in 3T3 cells. These cDNA clones were employed to screen the steady-state levels of cytoplasmic RNAs in a wide variety of viral (SV40, polyoma, adenovirus, and Rous sarcoma virus) and nonviral (methylcholanthrene, embryonal carcinoma) transformed cell lines. Two of the cDNA clones--A17 and 104--detected greater than 40-100-fold higher levels of mRNA in all the transformed cell lines tested when compared to nontransformed cells (3T3, C3HEF). The levels of mRNA complementary to these two cDNAs were regulated in a temperature-sensitive fashion (87-100-fold) in both SV40tsA- and RSV ts-src-transformed murine cell lines. These two cDNA clones detected greater than 100-fold, higher levels of complementary RNA derived from SV40 tumor tissue than in normal mouse liver. RNA species complementary to cDNA clones A17 or 104 were not detected in either actively growing nontransformed cells or in serum-stimulated 3T3 cells. The abundance levels of mRNAs detected by these two cDNA clones appear to be regulated 100-fold or greater by the transformed state, independent of the transforming agent. The higher levels of these RNA species detected in transformed mouse cells appear not to be solely regulated by the state of growth of nontransformed cells.


Stakeholder analysis: Medicare Diabetes Prevention Program awareness and implementation.
The Medicare Diabetes Prevention Program (MDPP) provides unprecedented coverage of a behavior change program for older adult Medicare beneficiaries, but uptake has been extremely limited; only 1.5 sites deliver the program per 100,000 beneficiaries nationwide. Inadequate reach and utilization of the MDPP threaten its long-term success; thus, the purpose of this project was to determine facilitators and barriers to MDPP implementation and use in western Pennsylvania. We conducted a qualitative stakeholder analysis project with suppliers of the MDPP and health care providers. Using an implementation science framework, we conducted individual interviews with 5 program suppliers and 3 health care providers (N = 8) to determine their perspectives on positive aspects of the program and reasons for MDPP unavailability and lack of use. Data were analyzed using Thorne and colleagues' approach of interpretive description. Three main themes emerged: (1) facilitators and attributes of the MDPP, (2) barriers to MDPP implementation, and (3) suggestions for improvement. Facilitators of the program included technical support and webinars from Medicare to assist with the application process. Barriers such as financial reimbursement constraints and a lack of a systematic referral process were noted. Stakeholders suggested refinements to participant eligibility and performance-based payments, a seamless method of flagging and referring patients through the electronic health record, and ongoing virtual program delivery options. Findings from this project can be used to improve implementation of the MDPP in western Pennsylvania, support Medicare policy refinement, and inform implementation research to promote broader adoption of the MDPP across the United States.


Gene delivery to the retina: from mouse to man.
With the recent progress in identifying disease-causing genes in humans and in animal models, there are more and more opportunities for using retinal gene transfer to learn more about retinal physiology and also to develop therapies for blinding disorders. Success in preclinical studies for one form of inherited blindness have led to testing in human clinical trials. This paves the way to consider a number of other retinal diseases as ultimate gene therapy targets in human studies. The information presented here is designed to assist scientists and clinicians to use gene transfer to probe the biology of the retina and/or to move appropriate gene-based treatment studies from the bench to the clinic.


The angiotensin II receptor blocker candesartan improves survival and mesenteric perfusion in an acute porcine endotoxin model.
Blockade of the angiotensin II type 1 (AT1) receptor has been demonstrated to ameliorate splanchnic hypoperfusion in acute experimental circulatory failure. This study focused on hemodynamic changes and survival in pigs treated with AT1 blockade prior to or during acute endotoxinemia. Escherichia coli lipopolysaccharide endotoxin was infused in anesthetized and mechanically ventilated pigs. Systemic, renal, mesenteric and jejunal mucosal perfusion as well as systemic oxygen and acid-base balance were monitored. The selective AT1 receptor blocker candesartan was administered prior to as well as during endotoxinemia. Control animals received the saline vehicle. Pre-treatment with candesartan resulted in higher survival rate (83%, 10 out of 12 animals) compared with 50% (6 of 12) in control animals and 27% (3 of 11) in animals treated during endotoxinemia. Pre-treatment with candesartan resulted in higher cardiac output, mixed venous oxygen saturation, arterial standard base-excess, portal venous blood flow during endotoxin infusion compared with controls and animals treated during endotoxinemia. No adverse effects were found on neither systemic nor renal circulation. The favorable results of AT1 receptor blockade prior to endotoxinemia are lost when blockade is established during endotoxinemia demonstrating the importance of the renin-angiotensin system and its dynamic involvement in acute endotoxinemic shock.


A critical examination of three-point bending for determining Young's modulus.
The three point bending method is commonly used for determining Young's modulus, E. However, the literature E values of some common pharmaceutical excipients, determined by three point bending, exhibit orders of magnitude variability that cannot be fully explained by normal variations in material properties. To understand the root cause for the large variations, we assessed several potential experimental factors, including instrument stiffness, data analysis method, test speed, alignment between loading beam and specimen surface, and ratio of sample thickness to distance between two supports. Our results show that E value is sensitive to 1) deformation of instrument under force, 2) force-displacement data analysis method, 3) misalignment between loading beam and specimen surface, 4) flashing at the edge of a specimen, 5) accuracy of the true density. Finally, strategies to improve the accuracy of E determination by the three point bending method were recommended.


[An impaired detection and clearance of dying cells can lead to the development of chronic autoimmunity].
Not properly cleared dead cells are dangerous for the body. The dead cells accumulate, lose their membrane integrity, danger signals are released, and nuclear antigens get accessible in an inflammatory context. In times of increased apoptosis, tolerance can be broken, a chronic inflammation results which then can lead to an autoimmune reaction against nuclear constituents. An impaired clearance of dying cells represents a central pathogenic process in the development of chronic autoimmune diseases like in systemic lupus erythematosus (SLE). Many adaptor molecules and receptors are involved in the clearance of dying cells. Complement components, serum DNase I, phosphatidylserine, and modified glycoproteins participate crucially in the clearance of apoptotic and necrotic cells. We further observed intrinsic defects of macrophages of some SLE patients. Macrophages as well as granulocytes of some SLE patients showed heterogeneous clearance defects. Furthermore, we observed an accumulation of nuclear material in germinal centres of lymph nodes of some SLE patients. The non-ingested nuclear material may provide survival signals for autoreactive B cells and consecutively antinuclear autoantibodies (ANA) will be produced. We therefore conclude that drugs promoting the phagocytosis are important candidates of specific therapies in the future which expect a more gentle and purposive treatment of patients with SLE.


Imatinib (STI-571) heals a gastrocutaneous fistula resulting from a malignant gastric stromal tumor.
The management of inoperable giant malignant gastrointestinal stromal tumor (GIST) of the stomach sed to be a formidable task. We report our success with the use of STI-571 in treating a patient with huge GIST of the stomach complicated by gastrocutaneous fistula after an unsuccessful laparotomy. The patient was a 49-year-old man who presented to our center with a painful epigastric mass in December 2001. Endoscopy, biopsy, and magnetic resonance scan confirmed that it was a malignant stromal tumor arising from the gastric fundus. Laparotomy with an intention to resect the tumor was performed in view of the obstructing symptoms. However, massive bleeding was encountered during dissection of the tumor and gastrectomy was abandoned. The case was further complicated with the development of a gastrocutaneous fistula in the early postoperative period. The patient ws then managed with naso-duodenal tube feeding and enteral STI-571 was prescribed. The fistula healed up in 20 days and the mass became impalpable 1 month afterwards. Follow-up computed tonography (CT) scan 3 months later confirmed significant tumor reduction, and the patient has experienced no side effects from the treatment


Early Results of Robotically Assisted Congenital Cardiac Surgery: Analysis of 242 Patients.
Robotic surgery is an alternative to traditional and minimally invasive cardiac procedures. The adaption of robotics in congenital cardiac surgery has remained limited, however. We analyzed the early outcomes of our single-center experience in robotically assisted congenital cardiac surgery. From May 2013 to February 2020, 242 robotic operations were done for secundum atrial septal defects (74.7%), sinus venous atrial septal defects (16.1%), partial anomalous pulmonary venous connections (10.7%), widely patent foramen ovale (3.7%), ostium primum defects (2.5%), unroofed coronary sinus (1.2%), partial atrioventricular canal defects (2.5%), residual septal defects after failed percutaneous closure (1.2%), ventricular septal defect (0.4%), cor triatriatum sinister (0.4%), subvalvular aortic stenosis (0.4%), common atrium (0.4%), and double-chambered right ventricle (0.4%) using the da Vinci system. There was no mortality. Mean age was 30.9 ± 12.1 years, and 132 (54.5%) patients were female. Thirty (12.3%) patients were pediatric (median, 16 years; range, 12-17 years). Mean cardiopulmonary bypass and aortic clamping times were 89.5 ± 30.0 and 44.9 ± 22.3 minutes, respectively. We noted a steady decrease in operation times during the first year. Conversion to larger thoracic incisions was needed in 2 (0.8%) patients. The postoperative rates of stroke, cardiac events, pulmonary complications, and reexploration were 0.4%, 2.4%, 4.1%, and 0.8%, respectively. Mean hospital stay was 3.5 ± 1.1 days. Robotic technology can be utilized to perform suitable congenital cardiac procedures safely and effectively. With acceptable complication rates, robotic surgery can be an alternative to traditional, minimally invasive, and endoscopic approaches in adolescent and adult patients.


Anatomical structure and nerve branching pattern of the human infraspinatus muscle.
The function of the infraspinatus muscle, critical to rotator cuff function, is dependent upon the muscle's structure and innervation pattern. The morphology of the infraspinatus muscle has been inconsistently described in the literature. Additionally, the branching pattern of the suprascapular nerve in the infraspinous fossa has not been addressed in the literature. The purposes of this study were to determine: the arrangement of the infraspinatus muscle bellies; the branching patterns of the suprascapular nerve to the infraspinatus muscle; if the infraspinatus muscle was composed the neuromuscular compartments. Forty-eight infraspinatus muscles from 24 embalmed cadavers were studied using standard dissection techniques to determine morphological characteristics and innervation patterns. Results demonstrated that the infraspinatus muscles were comprised of three separate muscular partitions with each partition residing in a thin fascial compartment but all residing deep to the posterior scapular fascia. A first order suprascapular nerve branch was present in 91.6% of superior, 100% of middle, and 70.8% of inferior partitions. A first order nerve was present in all 3 muscular compartments of the same infraspinatus muscle in 62.5% of cases. Second order nerve branches were present in 8.3% of superior, 0% of middle, and 29.2% of inferior partitions. These findings help to determine a more complete and accurate understanding of the structure of the infraspinatus muscle. A better understanding of its structure could lead to a better understanding of the function of the muscle. Such information will enable more effective rehabilitation strategies for injuries involving the infraspinatus component of the rotator cuff.


Evolution of prostate biopsy techniques. Looking back on a meaningful journey.
The technique of prostate biopsy has evolved a long way since its inception to being a safe diagnostic procedure. The principles of the biopsy technique continue to improvise with the knowledge about prostate cancer and availability of newer treatment options like active surveillance and focal therapy. Currently, we depend on accurate cancer information from the biopsy more than ever for deciding the ideal treatment option. The aim of this review is to present the major milestones in prostate biopsy technique evolutions and its impact on the prostate cancer management. We performed a detailed non-systematic literature review to present the historical facts on the transformations in prostate biopsy techniques and also the direction of present research to improve accurate cancer detection. There is a clear change in trend in biopsy technique before and after the introduction of transrectal ultrasound and prostate specific antigen. In the earlier era, the biopsies were aimed at palpable nodules and obtaining adequate prostatic tissue for diagnosis while the later era has moved towards detection of non-palpable and early prostate cancer. Recently, there is an increasing trend towards image guided targeted biopsies to extract maximum cancer information from minimum biopsy cores. Prostate biopsy techniques have seen major changes since its inception and have a major impact on prostate cancer management. There is a great potential for research which can further support the newer treatment options like focal therapy.


[The effect of trisporic acids on the composition of components of the cell wall in Blakeslea trispora Thaxter].
The cell wall of Blakeslea trispora was found to contain chitin similar to crustaceaous chitin in physico-chemical properties; this was confirmed by the data of IR spectroscopy and X-ray diffraction analysis. Trisporic acids, hormonal regulators of reproduction, hardly affected the content of chitin, xylan, and other polysaccharides of the cell wall of Blakeslea trispora, but increased the content of protein.


The decision to operate: role of integrated computed tomography positron emission tomography in staging oesophageal and oesophagogastric junction cancer by the multidisciplinary team.
Our objective was to assess the role of fusion positron emission tomography-computed tomography (PET-CT) in staging patients for minimally invasive oesophagectomy (MIO) with potentially resectable disease from the perspective of a multidisciplinary team (MDT) deciding on operability with conventional staging investigations. Fifty consecutive patients presenting with potentially operable oesophageal or oesophagogastric junctional tumours were staged with computed tomography (CT) and endoluminal ultrasound (EUS). The MDT categorised patients as group A (n=33; CT N0M0) or group B (n=17; CT N1/possible M1). All patients underwent FDG PET-CT. Patients with localised disease (at T3), including single level N1 disease on PET-CT, were deemed suitable for induction chemotherapy followed by surgery. PET-CT re-categorised 12% of patients as inoperable on grounds of distant metastases (four in group A, two in group B). Five patients did not proceed to resection for other reasons. Two had metastatic disease at thoracoscopy. Resection specimens (n=37) contained 24 nodes (median). Compared with pN status, positive predictive value of PET-CT was 40% and negative predictive value was 43%. The expected PET-CT N1 group had the highest mean number of involved nodes. Median survival for all patients (n=50) was 31.9 months for group A compared with 17.3 months for group B (not statistically significant). There was no significant difference between patients who were PET-CT N0 or N1 in survival or disease-free survival in patients undergoing surgery (n=37). PET-CT informs the MDT decision to operate in avoiding futile surgery in stage IV disease or widespread nodal disease. In this study, overall PET-CT N1 status has low positive and negative predictive value for overall pN status.


TNFSF4 is a risk factor to systemic lupus erythematosus in a Latin American population.
The aim of this study was to examine the association of three TNFSF4 single nucleotide variants (SNVs) with systemic lupus erythematosus (SLE) susceptibility in Mexican patients. Genotypes of the TNFSF4 rs1234315T/C, rs2205960G/T, and rs704840T/G SNVs were determined using a TaqMan assay. In our study, we included 395 patients with SLE and 500 controls. Our information shows a significant difference in the allelic and genotypic frequency of the three TNFSF4 SNVs between cases and controls. Thus, our data showed an association between TNFSF4 rs1234315T/C (T vs. C, OR 1.40, p = 0.00087), rs2205960G/T (G vs. T, OR 1.32, p = 0.0037), and rs704840T/G (T vs. G, OR 1.41, p = 0.0003) and SLE susceptibility in Mexican subjects. Besides, we conducted a meta-analysis to determine the role of TNFSF4 rs2205960G/T and SLE susceptibility; our results showed that this variant is a risk factor for SLE in Latin Americans and Asians. Our results show that TNFSF4 rs1234315T/C, rs2205960G/T, and rs704840T/G are risk factors to SLE in Mexicans. This is the first study to document an association between TNFSF4 rs704840T/G and SLE in a Latin American population. In addition, our meta-analysis showed that TNFSF4 rs2205960G/T is a risk factor for Asians and Latin Americans. Key Point • The TNFSF4 rs1234315T/C, rs2205960G/T, and rs704849T/G SNVs are risk factors to SLE in patients from Mexico.


Training aid for mental care staff.
A training pack to help mental health professionals challenge discrimination in health services was launched last week.


Effects of seed distribution and competitors on seed harvesting efficiency in heteromyid rodents.
The foraging strategies of four naturally co-existing heteromyid rodent species were investigated: Dipodomys deserti (≈100 g), D. merriami (≈38 g), Microdipodops pallidus (≈13 g), and Perognathus longimembris (≈7 g). In 208 over-night laboratory foraging trials animals were provided with millet seed distributed in clumped and scattered patterns. Net removal of seeds from the foraging arena and amounts of seeds in cheek pouches and in caches were determined. When alone in an areana none of these species specialized extensively on either clumped or scattered seeds, although each tended to take more clumped than scattered seeds. When placed together with other individuals, animals once again tended to cache more clumped than scattered seeds in all but one paired combination of species: P. longimembris cached more scattered than clumped seeds when opposed by D. deserti. This suggests that the smaller species obtained a less preferred distribution of seeds in the face of competition. The two smaller species showed a great reduction in general foraging success in the presence of either of the two larger species. In general, a species cached less seeds when faced by larger opponent species.


A rare complication of intraabdominal hydatid disease: gastric fistula and recurrent gastric bleeding.
Intra-abdominal hydatid disease is a type of echinococcosis. This silent disease becomes symptomatic with serious complications. A 50-year-old woman presented with abdominal pain and recurrent hematemesis. A computed tomographic scan demonstrated a large retrogastric hydatid cyst and suggested gastric fistula. An emergent laparotomy was performed, and gastric fistula of the intraperitoneal hydatid cyst and arterial bleeding around the fistula were demonstrated intraoperatively. Clinical course, complications, and management of intra-abdominal hydatid disease are discussed.


A system model for investigating passive electrical properties of neurons.
Passive membrane properties of neurons, characterized by a linear voltage response to constant current stimulation, were investigated by busing a system model approach. This approach utilizes the derived expression for the input impedance of a network, which simulates the passive properties of neurons, to correlate measured intracellular recordings with the response of network models. In this study, the input impedances of different network configurations and of dentate granule neurons, were derived as a function of the network elements and were validated with computer simulations. The parameters of the system model, which are the values of the network elements, were estimated using an optimization strategy. The system model provides for better estimation of the network elements than the previously described signal model, due to its explicit nature. In contrast, the signal model is an implicit function of the network elements which requires intermediate steps to estimate some of the passive properties.


Long-and short-range functional connectivity density alteration in non-alcoholic cirrhotic patients one month after liver transplantation: A resting-state fMRI study.
To longitudinally evaluate long- and short-range functional connectivity density (FCD) alteration in cirrhotic patients one month after liver transplantation (LT) and their correlation with cognitive changes by using resting-state functional magnetic resonance imaging (rs-fMRI). Twenty seven candidates awaiting LT and 24 age-, gender-, and education-matched healthy controls (HCs) were studied. All 27 patients and HCs performed rs-fMRI examinations. Of 27 cirrhotic patients, 13 patients received LT underwent the repeated rs-fMRI examinations one month after LT. Laboratory and psychometric tests were carried out. The long- and short-range FCD maps derived from degree centrality calculation were compared. Correlations between FCD alteration and laboratory/psychometric changes were evaluated as well. In cirrhotic patients, most of the brain areas with altered long- and short-range FCD could reverse one month after LT, which was accompanied with cognitive and liver functional improvement. The reduced long-range FCD in right posterior cingulate cortex (PCC) and Left middle frontal gyrus (MFG), and reduced short-range FCD in right precuneus (PCu) persisted in the early period after LT. In addition, one month after LT, the post-LT group showed reduced long-range FCD in right rectus gyrus (REC) and left medial prefrontal cortex (MPFC), and reduced short-range FCD in left middle temporal gyrus (MTG), when compared with the pre-LT group. We found Δdigital symbol test (ΔDST) score positively correlated with long-range ΔFCD in right precentral gyrus (preCG) (r = 0.72, P < 0.01) and right supplementary motor area (SMA) (r = 0.59, P < 0.05). LT results in favorable effect on cognitive function in cirrhotic patient, which can be reflected by FCD alteration. However, persistence of PCC/PCu functional connectivity disturbance one month after LT indicates complete cognitive function restoration may need a longer time.


Influence of soil characteristics on rare earth fingerprints in mosses and mushrooms: Example of a pristine temperate rainforest (Slavonia, Croatia).
The present study aims to investigate levels and distribution of rare earth elements (REE) in soils, mosses and mushrooms of a pristine temperate rainforest, a non-polluted natural system, in order to characterise their environmental availability and mobility. The multielement analysis of digested soil, moss and mushroom samples was performed by High Resolution Inductively Coupled Plasma Mass Spectrometry. The distribution of rare earths in mosses and mushrooms was found primarily affected by local pedological setting. Mosses displayed a consistent lithological signature with an almost insignificant REE fractionation compared to soils. Mushrooms showed differences in REE concentrations in certain parts of the fruiting body with regard to their main physiological function and indicated a significant impact of soil organic content on the overall REEs uptake. Results of our work highlight the importance of substrate characteristics on the initial levels of REEs in mosses and mushrooms. Moreover, this study provides baseline data on the rare earth element levels in mosses and mushrooms growing in a pristine forest area characterised by naturally elevated REE levels in the soil.


Ion chromatography for determination of nitrite and nitrate in seawater using monolithic ODS columns.
A fast and highly sensitive ion chromatographic method using monolithic ODS columns was developed for the determination of nitrite (NO2-) and nitrate (NO3-) in seawater. Two monolithic ODS columns (50 mm x 4.6 mm i.d. + 100 mm x 4.6 mm i.d.) connected in series were coated and equilibrated with 5 mM cetyltrimethylammonium chloride (CTAC) aqueous solution. The column efficiency with 0.5 M NaCl as the mobile phase did not decrease in spite of the increase in flow rate of the mobile phase. Thus, good chromatograms were obtained within 3 minutes for NO2- and NO3 in artificial seawater without interferences by coexisting ions. The detection limit (S/N = 3) with UV detection at 225 nm was 0.8 and 1.6 microg/L for NO2- and NO3-, respectively. The characteristics of the monolithic CTA(+)-coated ODS columns were discussed. The present method was successfully applied to the fast and sensitive determination of NO2- and NO3- in real seawater samples.


Experimental estimation of the viscous component of ultrasound attenuation in suspensions of bovine skeletal muscle myofibrils.
It has been suggested that viscous losses, caused by the motion of myofibrils relative to their environmental fluid, could be the major cause of ultrasound attenuation in muscle. This Paper presents theoretical and experimental estimates of the viscous component of attenuation in suspensions of myofibrils. Experimental estimates were made by monitoring the effects of varying the viscosity and density of the suspending fluid and ranged from 0.55 to 0.72 cm2 g-1 protein at 7 MHz, when expressed as mass attenuation coefficients. This represented 16-22% of the total attenuation measured in suspensions. Corresponding theoretical calculations were lower: 0.44 and 0.21 cm2 g-1 protein, respectively, for suspensions with the myofibrils aligned along and across the direction of sound propagation. It was concluded that most of the attenuation was caused by other absorption processes.


Mechanisms of protein degradation: an odyssey with ODC.
Intracellular proteolysis plays an important role in regulating fundamental cellular processes such as cell cycle, immune and inflammation responses, development, differentiation, and transformation. The ubiquitin-proteasome system accounts for the degradation of the majority of cellular short-lived proteins. This system involves the conjugation of multiple ubiquitin residues to the target protein and its recognition by the 26S proteasome through the poly-ubiquitin chain. Studies on the degradation of ornithine decarboxylase (ODC) demonstrated that poly-ubiquitin is not the only signal recognized by the 26S proteasome. The recognition of ODC by the 26S proteasome is mediated by interaction with a polyamine-induced protein termed, antizyme (Az). While the degradation of ODC is ubiquitin-independent, the degradation of its regulator Az, and of antizyme-inhibitor (AzI), an ODC homologous protein that regulates Az availability, are ubiquitin dependent. Interestingly, ODC undergoes another type of ubiquitin-independent degradation by the 20S proteasome that is regulated by NAD(P)H quinone oxidoreductase 1 (NQO1). Considering the prevalence of the ubiquitin system in the process of cellular protein degradation it is rather remarkable that a key cellular enzyme is subjected to two different proteolytic pathways that are different from the ubiquitin dependent one. This exceptional behavior of ODC provides us with valuable insights regarding protein degradation in general.


Studies on the effect of monosodium glutamate [MSG] administration on some antioxidant enzymes in the arterial tissue of adult male mice.
The subcutaneous administration of monosodium glutamate to normal adult male mice at dose levels of 4 and 8 mg/g body weight caused a significant increase in lipid peroxidation level in the arterial tissue. The levels of total sulfhydryl and protein-bound sulfhydryl groups were found to be significantly increased, whereas non-protein-bound sulfhydryl, representing the glutathione level, was significantly decreased. It was also observed that the administration of monosodium glutamate at a dose level of 4 mg/g body weight and above induced oxidative stress by significantly lowering the activities of antioxidant enzymes like superoxide dismutases, catalase, and glutathione metabolizing enzymes like glutathione reductase and glutathione peroxidase in the arterial tissue.


Chlorine dioxide reaction with selected amino acids in water.
Chlorine dioxide is a hypochlorite alternative disinfectant agent. In this context, we have determined the products formed in the reaction of ClO(2) with selected amino acids as model compounds that can be present in natural waters. The reaction of tryptophane, histidine and tyrosine (10 ppm each) with ClO(2) were studied at molar ratios ranging from 0.25 to 4 in the presence or absence of oxygen. It was found that in the absence of oxygen adding substoichiometric amounts of ClO(2) creates products that are structurally similar to the starting amino acids. Through a series of cascade reactions the initial product distribution gradually evolves toward simple, small carbon chain products that are far from the starting amino acid. The reaction product distribution revealed that chlorine dioxide can attack the electron-rich aromatic moieties as well as the nitrogen atom lone electron pair. Our study is relevant to gain knowledge on the reaction mechanism of ClO(2) with ubiquitous amino acids present in natural waters.