Noninvasive oxygen partial pressure measurement of human body fluids in vivo using magnetic resonance imaging.
The oxygen partial pressure (pO2) of human body fluids reflects the oxygenation status of surrounding tissues. All existing fluid pO2 measurements are invasive, requiring either microelectrode/optode placement or fluid removal. The purpose of this study is to develop a noninvasive magnetic resonance imaging method to measure the pO2 of human body fluids. We developed an imaging paradigm that exploits the paramagnetism of molecular oxygen to create quantitative images of fluid oxygenation. A single-shot fast spin echo pulse sequence was modified to minimize artifacts from motion, fluid flow, and partial volume. Longitudinal relaxation rate (R1 = 1/T1) was measured with a time-efficient nonequilibrium saturation recovery method and correlated with pO2 measured in phantoms. pO2 images of human and fetal cerebrospinal fluid, bladder urine, and vitreous humor are presented and quantitative oxygenation levels are compared with prior literature estimates, where available. Significant pO2 increases are shown in cerebrospinal fluid and vitreous following 100% oxygen inhalation. Potential errors due to temperature, fluid flow, and partial volume are discussed. Noninvasive measurements of human body fluid pO2 in vivo are presented, which yield reasonable values based on prior literature estimates. This rapid imaging-based measurement of fluid oxygenation may provide insight into normal physiology as well as changes due to disease or during treatment.


Reliability of QRS duration and morphology on surface electrocardiogram to identify ventricular dyssynchrony in patients with idiopathic dilated cardiomyopathy.
Using Fourier phase analysis of equilibrium radionuclide angiography in both ventricles, inter- and intraventricular delays were correlated with QRS morphology and duration in 103 patients with idiopathic dilated cardiomyopathy. We found that left bundle branch block on the left axis may reflect a high intra-left ventricular dyssynchrony. A precise evaluation of the dyssynchrony appears useful because a high proportion of patients with incomplete bundle branch block, left anterior hemiblock, or "normal" QRS exhibit a marked intra-LV dyssynchrony and may respond to cardiac resynchronization therapy.


Integrating smartphone technology, social support and the outdoor built environment to promote community-based aerobic and resistance-based physical activity: Rationale and study protocol for the 'ecofit' randomized controlled trial.
Regular physical activity can significantly reduce the risk of numerous chronic diseases, and improve bone density and mental health. Yet, only 50% of Australian adults meet the aerobic physical activity guidelines and 9-19% meet the resistance-based physical activity guidelines. The aim of this study is to enhance community-based aerobic and resistance-based physical activity through the use of publicly available outdoor exercise equipment, social support and smartphone technology. The ecofit intervention will be evaluated using a two-arm randomized controlled trial. A total of 240 adults (aged 18-80) will be recruited and randomly allocated to either the ecofit intervention or a 'wait-list' control group. Both groups will have access to the two types of outdoor park exercise equipment, but the intervention group will be given access to the purpose-built ecofit app and a 90-min introductory group training session. To promote social support, participants can enrol in a group of up to four individuals and access the ecofit Facebook group. The ecofit app include workout plans that can be tailored to different locations, difficulty levels and workout-types (i.e., resistance-only or combined resistance and aerobic workouts). Outcome assessments will be conducted at baseline, 3- (primary-end point) and 9-months follow-up. The primary outcomes are upper and lower body muscular fitness. The secondary outcomes include physical activity, body composition, aerobic fitness, body mass index, self-report resistance-based physical activity, and mental health outcomes. The cost-effectiveness of the study will also be evaluated. ecofit is an innovative, multi-component physical activity intervention that integrates smartphone technology, social support and the outdoor built environment to promote community-based aerobic and resistance-based physical. The findings will be used to guide future interventions and to support councils to promote community-based physical activity through the use of local outdoor exercise equipment. ACTRN12619000868189.


Dietary Antioxidants and Cardiovascular Health-Editorial Comments and Summary.
The influence of dietary antioxidants on health is supported by a large body of evidence [...].


The potential of Mikania micrantha (Chinese creeper) to hyperaccumulate heavy metals in soil contaminated by electronic waste.
The objective of this study is to investigate the contributions of Mikania micrantha (chinese creeper) to remediate heavy metal pollutants present in an e-waste-contaminated soil. Different proportions of e-waste soil (0%, 25%, 50%, 75%, and 100% w/w) planted with Cynodon dactylon together with the test species were prepared for testing relative seed germination (RSG), relative root growth (RRG), and germination index (GI) tests. A significant higher value of GI (77.8%) was found in M. micrantha than that of the other species when planting in 100% e-waste-contaminated soil. A significant correlation (< 0.05) was found between heavy metal concentration and germination assays in M. micrantha. A significant decrease in heavy metal concentration of the polluted soil after the experiment indicated that biomolecule development studies to determine the aggregate benefit of M. micrantha for phytoremediation remain to be studied in future.


Phenotypic characterization of normal human placental mononuclear cells.
The placenta is a rich source of immunocompetent cells. We have studied the phenotype, number and origin of placental mononuclear blood cells isolated from 32 normal term placentae using 4 color flow cytometry. Respective maternal and cord blood leucocyte preparations were also compared. Placental tissue without extraembryonic membranes was cut into small pieces and divided. One portion was washed extensively with ice-cold PBS. Both tissue portions were disrupted in a blender and cells were dissociated by using a 180 mu sieve. Leucocytes were isolated by Ficoll-Hypaque density gradient centrifugation. Maternal and cord bloods were HLA typed and in cases of HLA-A2 or B7/40 disparity, monoclonal anti-HLA antibodies to these antigens showed that unwashed placental tissue contained 35% maternal and 65% fetal cells. This ratio, however, was not reflected for a given cell phenotype. In comparison, washed placental tissue contained cells of fetal origin only. Both unwashed and washed placental tissue contained fewer CD3 and CD4, but more CD8 cells than maternal and cord blood. Markers of NK cells such as, CD16, CD56, and CD57 showed this cellular phenotype to be 15 times more abundant in the placental preparations than in cord and maternal blood. The quantitative differences between peripheral blood and placental CD8 and NK cells were further explored with an antiprogesterone receptor antibody in combination with anti-CD8, anti-CD57 and anti-HLA-DR. The number of progesterone receptor (PGR) positive cells was three times higher in placental tissues than in cord or maternal blood. These data indicate that the phenotypic frequencies of certain placental leucocytes are significantly different from maternal and fetal peripheral blood. Progesterone and the presence of PGR may be important in the differential retention of placental leucocytes.


Effects of dietary zinc levels, phytic acid and resistant starch on zinc bioavailability in rats.
Owing to its fermentability, it has been advocated that resistant starch (RS) has a positive effect on the absorption of minerals by increasing their solubility in the hindgut. In marginally zinc-deficient rats, the enhancement of zinc bioavailability by RS occurs mostly when the diet contains phytic acid. This study aims to investigate the effect of dietary zinc level and phytic acid on the cecal zinc pools and zinc bioavailability of rats fed RS. Wistar rats (male, 3wk old) were divided into eight groups (n = 6), and fed diets containing either 5% cellulose (control fiber: insoluble and low fermentable) or 20 % RS (test fiber: soluble and fermentable), with or without the addition of 1% sodium phytate, at the 10 and 30 mg/kg dietary zinc levels, for 21 days. At 10 mg Zn/kg, RS increased femur zinc concentration only in the group receiving the phytate-containing diet, while at 30 mg Zn/kg it increased femur zinc concentration in rats fed both phytate-free and phytate-containing diets. The total content of zinc in the cecum was increased by the higher dietary zinc level and tended to be increased by the addition of phytate, which is assumed to impair zinc absorption in the small intestine. Feeding RS lowered cecal pH values, which correlated with increasing values of zinc solubility (r = -0.3471; P < 0.05). The later was, in turn, directly associated with zinc apparent absorption (r = 0.3739; P < 0.05). The increase in zinc bioavailability by RS occurs when dietary zinc levels are adequate and/or zinc absorption is impaired in the small intestine, increasing the influx of unabsorbed zinc into the cecum and favoring the increase of zinc bioavailability when RS fermentation lowers the cecal pH.


Smartphone-based cough monitoring as a near real-time digital pneumonia biomarker.
Cough represents a cardinal symptom of acute respiratory tract infections. Generally associated with disease activity, cough holds biomarker potential and might be harnessed for prognosis and personalised treatment decisions. Here, we tested the suitability of cough as a digital biomarker for disease activity in coronavirus disease 2019 (COVID-19) and other lower respiratory tract infections. We conducted a single-centre, exploratory, observational cohort study on automated cough detection in patients hospitalised for COVID-19 (n=32) and non-COVID-19 pneumonia (n=14) between April and November 2020 at the Cantonal Hospital St Gallen, Switzerland. Cough detection was achieved using smartphone-based audio recordings coupled to an ensemble of convolutional neural networks. Cough levels were correlated to established markers of inflammation and oxygenation. Cough frequency was highest upon hospital admission and declined steadily with recovery. There was a characteristic pattern of daily cough fluctuations, with little activity during the night and two coughing peaks during the day. Hourly cough counts were strongly correlated with clinical markers of disease activity and laboratory markers of inflammation, suggesting cough as a surrogate of disease in acute respiratory tract infections. No apparent differences in cough evolution were observed between COVID-19 and non-COVID-19 pneumonia. Automated, quantitative, smartphone-based detection of cough is feasible in hospitalised patients and correlates with disease activity in lower respiratory tract infections. Our approach allows for near real-time telemonitoring of individuals in aerosol isolation. Larger trials are warranted to decipher the use of cough as a digital biomarker for prognosis and tailored treatment in lower respiratory tract infections.


Three-Dimensional Regulation of Ferroptosis at the Intersection of Iron, Sulfur, and Oxygen Executing Scrap and Build Toward Evolution.
Significance: Iron is an essential element for every life on earth as a primary media for electron flow. Sulfur compounds as sulfhydryls counteract catalytic activity of iron whereas sulfur overdose is also toxic. In aerobic organisms, oxygen is the major media for electron transfer with higher intracellular mobility, which cooperates with the iron system. Based on the importance of iron, there is no active pathway to excrete iron outside the body in higher species. Whereas bacterial infection causes a scramble for iron in situ, cancer can be the outcome of the side effects of long use of iron and oxygen. Recent Advances: Ferroptosis is a recently coined cell death, defined as catalytic Fe(II)-dependent regulated necrosis accompanied by lipid peroxidation. Researchers recently recognized that ferroptosis is involved in a variety of physiological and pathological contexts, including embryonic erythropoiesis, aging, neurodegeneration and cancer cell death. Alternatively, carcinogenesis is a process to obtain iron addiction with ferroptosis-resistance, based on rodent animal studies. Critical Issues: Here we propose that ferroptosis is three-dimensionally regulated by iron, sulfur and oxygen, which correspond to oxidants, antioxidants and membrane fluidity with susceptibility to lipid peroxidation, respectively. Future Directions: Whereas life attempts to prevent ferroptosis, ferroptotic cells eventually emit iron-loaded ferritin as extracellular vesicles to maintain monopoly of iron. Antioxid. Redox Signal. 39, 807-815.


Behaviour-based pain scales: Validity and interrater reliability of BPS-NI and PAINAD-G on general wards.
Measuring pain intensity in patients unable to communicate is a challenge on general wards. Observation-based pain scoring tools have been used for patients with dementia or in critically ill intensive care unit (ICU) patients. However, there is no established or validated assessment tool for non-ICU patients without dementia related cognitive deficiencies who cannot communicate. The "Behavioural Pain Scale Non-Intubated" (BPS-NI) and the "Pain Assessment In Advanced Dementia-German" (PAINAD-G) are potential tools to fill this gap. This study was performed with verbal non-ICU patients on general wards at Charité Berlin. Two assessors independently rated pain intensity using the BPS-NI and the PAINAD-G along with patients' self-ratings on the Numeric Rating Scale (NRS). The interrater-reliability of BPS-NI and PAINAD-G was calculated and ROC-analyses were performed to identify cut-off values for medium and intense pain for each score. Effectiveness was calculated using percentage agreement. In total, 126 patients were included into analysis. The BPS-NI showed substantial congruence in interrater-reliability (Cohens-Kappa 0.71), whereas the PAINAD-G showed moderate congruence (Kappa 0.48). Based on ROC-analyses, for medium pain levels a cut-off 4 (BPS-NI) or 2 (PAINAD-G) and for severe pain levels cut-off 5 (BPS-NI) and 3 (PAINAD-G) would result in good accordance with self-reported NRS for pain. The BPS-NI shows a good validity in measuring pain intensity in patients on general wards and may possibly be used for patients unable to communicate. Using defined cut-off values for BPS-NI and PAINAD-G, clinically relevant pain intensities in patients can reliably be detected. Measuring the pain intensity in patients unable to communicate is a common challenge on general wards. This study assessed reliability and applicability BPS-NI and the PAINAD-G in a general ward setting. Furthermore, it provides cut-off values in order to estimate pain intensity and support analgesic response.


Schistosoma mansoni: melatonin enhances efficacy of cercarial and soluble worm antigens in the induction of protective immunity against infection in the hamster.
Based on the beneficial influence of melatonin administration on the course of schistosomiasis and on its possible action on the immune system, we aimed in this study to establish an immunization program using Schistosoma mansoni adult worm antigen (SWAP) and cercarial antigen (CAP) alone or concurrently with melatonin treatment, for 30 successive days, in an attempt to enhance their efficacy against the infection in hamsters. Results showed that the worm reduction percentages were 53.8%, 67.01%, 56.4% and 99.3% for CAP, CAP+melatonin, SWAP, SWAP+melatonin, respectively, indicating that melatonin enhanced efficacy of SWAP but only produced a slight increase in efficacy of CAP. Highly significant reductions in egg load in the liver and alteration in the oogram pattern with a high percentage of immature eggs and few dead eggs were recorded in the groups that received melatonin treatment suggesting a possible role for melatonin in the regulation of egg production and development. On the other hand, melatonin clearly improved the oxidative status in the immunized groups. No antibody (Ab) response was recorded in the groups immunized with SWAP+melatonin while low Ab level was seen in the other melatonin-treated group. In addition to the antioxidant properties of melatonin, our results suggested that the early and continuous melatonin administration may result in immunomodulatory actions which in turn enhanced the efficacy of SWAP and CAP in different ways. This indicates the importance of further investigation of the mechanisms of melatonin action and the possible application in a vaccination program.


An autecological study of Palicourea riparia Bentham as related to rain forest disturbance in Puerto Rico.
The present study is an attempt to understand biology of Palicourea riparia Bentham relative to the process of recovery following disturbance in a tropical rain forest in Puerto Rico. In this species seed germination and seedling maintenance are favored by environmental conditions associated with disturbance, yet germination and seedling persistence are significant in undisturbed forest conditions. The pattern of seed germination indicates that (1) the seeds possess a dormancy mechanism, (2) dormany is not evenly distributed within a seed population, and (3) some seeds can remain viable if buried in the soil for at least three years. Results from reciprocal transplant studies indicate that there has been no differentiation into growth ecotypes in this species. Photosynthetic rates are very low under both field and laboratory conditions. In the laboratory, photosynthetic rates are consistently enhanced by increases in light intensity. It is concluded that P. riparia is a species of wide ecological amplitude that acts as a gap opportunist.


Molecular determinants of inhibition of UCP1-mediated respiratory uncoupling.
Brown adipose tissue expresses uncoupling protein 1 (UCP1), which dissipates energy as heat, making it a target for treating metabolic disorders. Here, we investigate how purine nucleotides inhibit respiration uncoupling by UCP1. Our molecular simulations predict that GDP and GTP bind UCP1 in the common substrate binding site in an upright orientation, where the base moiety interacts with conserved residues R92 and E191. We identify a triplet of uncharged residues, F88/I187/W281, forming hydrophobic contacts with nucleotides. In yeast spheroplast respiration assays, both I187A and W281A mutants increase the fatty acid-induced uncoupling activity of UCP1 and partially suppress the inhibition of UCP1 activity by nucleotides. The F88A/I187A/W281A triple mutant is overactivated by fatty acids even at high concentrations of purine nucleotides. In simulations, E191 and W281 interact with purine but not pyrimidine bases. These results provide a molecular understanding of the selective inhibition of UCP1 by purine nucleotides.


Influence of exogenous phytase supplementation on phytate degradation, plasma inositol, alkaline phosphatase, and glucose concentrations of broilers at 28 days of age.
Inositol is the final product of phytate degradation, which has the potential to serve as an indicator of phytase efficacy. An experiment was conducted to evaluate effects of supplementing broiler diets with phytase on phytate degradation and plasma inositol concentrations at 28 d of age. Twenty-four Ross × Ross 708 male chicks were placed in battery cages (4 birds per cage) from 1 to 21 d of age and individually from 22 to 28 d of age. At 27 d of age, a catheter was placed in the brachial vein of broilers to avoid repeated puncture of the vein during blood collection. At 28 d of age, broilers received 1 of 3 experimental diets formulated to contain 0, 400, or 1,200 phytase units (FTU)/kg, respectively, in diet 1, 2, and 3. Blood was collected 1 h before feeding experimental diets and from 20 to 240 min after feeding experimental diets at 20-min intervals with a final blood collection at 480 min to determine plasma inositol concentrations. Inositol phosphate (IP) ester degradation was determined in gizzard contents and ileal digesta. Broilers provided the 1,200 FTU/kg phytase diet had 60% less (P < 0.01) IP6 concentration in gizzard content (1,264 vs. 4,176 nmol/g) and ileal digesta (13,472 vs. 33,244 nmol/g) than birds fed the 400 FTU/kg diet. Adding phytase at 1,200 FTU/kg increased (P < 0.01) inositol concentrations in gizzard content and ileal digesta of broilers by 2.5 (2,703 vs. 1,071 nmol/g) and 3.5 (16,485 vs. 4,667 nmol/g) fold, respectively, compared with adding 400 FTU/kg. Plasma inositol concentration of broilers was not different (P = 0.94) among the dietary treatments at each collection time. Inositol liberation in the digesta of broilers fed diets with 1,200 FTU/kg phytase did not translate to increased plasma inositol concentrations, which warrants further investigation.


The contrast-enhanced CT scan and the radionuclide brain scan: parallel mechanisms of action in the detection of supratentorial astrocytomas.
The preoperative contrast-enhanced CT scan and the radionuclide brain scan of 70 patients with surgically verified supratentorial astrocytomas were evaluated and compared. The results indicate parallel mechanisms of action of contrast enhancement and radionuclide uptake. These diagnostic modalities apparently mirror the integrity of the blood-brain barrier (BBB) and therefore are useful in assessing the degree of malignancy of supratentorial gliomas. However, lesions with an intact BBB will be missed with RN imaging. These lesions and/or the associated mass effect will be detected with contrast-enhanced computed tomography. Our findings firmly establish contrast-enhanced CT as the primary investigate tool in the suspected brain tumor.


Liver X Receptors as Therapeutic Targets for Managing Cholesterol: Implications for Atherosclerosis and Other Inflammatory Conditions.
Atherosclerosis is a disease characterized by excess cholesterol and inflammation in the blood vessels. The liver X receptors (alpha and beta) are members of the nuclear hormone receptor family that are activated by endogenous cholesterol metabolites. These receptors are widely expressed with a tissue distribution that includes the liver, intestine and macrophage. Upon activation, these receptors have been shown to increase reverse cholesterol transport from the macrophage back to the liver to aid in the removal of excess cholesterol. More recently, they have also been shown to inhibit the inflammatory response in macrophages. These functions are accomplished through direct regulation of gene transcription. Herein, we will describe the key benefits and potential risks of targeting the LXRs for the treatment of atherosclerosis.


Standardization of medium composition and agricultural waste in the production of p-hydroxybenzoic acid by Paecilomyces variotii.
Biotransformation of p-coumaric acid by Paecilomyces variotii resulted in the accumulation of a metabolite which was identified as p-hydroxybenzoic acid. We studied the possibility of enhancing this metabolite by various parameters such as varying the concentration of substrate, addition of different nitrogen sources and carbon sources at varying concentration and use of different agricultural wastes as a nutritive medium. Maximum amount of 254.6 ± 9.34 mg/kg of p-hydroxybenzoic acid was detected in the solid medium of corncob on 4th day of incubation using 10 mM p-coumaric acid as a sole carbon source. Consequently, P. variotii can be used as candidate fungi for the production of p-hydroxybenzoic acid. Solid wastes can be managed by using them for biotransformation of pharmaceutically important phenolic acids and hence cut down the cost of production of value-added compounds using a low-cost medium.


Urinary tract infection in children: Diagnosis, treatment, imaging - Comparison of current guidelines.
Urinary tract infection (UTI) is a frequent disorder of childhood, yet the proper approach for a child with UTI is still a matter of controversy. The objective of this study was to critically compare current guidelines for the diagnosis and management of UTI in children, in light of new scientific data. An analysis was performed of the guidelines from: American Academy of Pediatrics (AAP), National Institute for Health and Care Excellence (NICE), Italian Society of Pediatric Nephrology, Canadian Paediatric Society (CPS), Polish Society of Pediatric Nephrology, and European Association of Urology (EAU)/European Society for Pediatric Urology (ESPU). Separate aspects of the approach for a child with UTI, including diagnosis, treatment and further imaging studies, were compared, with allowance for recent research in each field. The analyzed guidelines tried to reconcile recent reports about diagnosis, treatment, and further diagnostics in pediatric UTI with prior practices and opinions, and economic capabilities. There was still a lack of sufficient data to formulate coherent, unequivocal guidelines on UTI management in children, with imaging tests remaining the main area of controversy. As a result, the authors formulated their own proposal for UTI management in children.


The influence of hierarchy on probability judgment.
Consider the task of predicting which soccer team will win the next World Cup. The bookmakers may judge Brazil to be the team most likely to win, but also judge it most likely that a European rather than a Latin American team will win. This is an example of a non-aligned hierarchy structure: the most probable event at the subordinate level (Brazil wins) appears to be inconsistent with the most probable event at the superordinate level (a European team wins). In this paper we exploit such structures to investigate how people make predictions based on uncertain hierarchical knowledge. We distinguish between aligned and non-aligned environments, and conjecture that people assume alignment. Participants were exposed to a non-aligned training set in which the most probable superordinate category predicted one outcome, whereas the most probable subordinate category predicted a different outcome. In the test phase participants allowed their initial probability judgments about category membership to shift their final ratings of the probability of the outcome, even though all judgments were made on the basis of the same statistical data. In effect people were primed to focus on the most likely path in an inference tree, and neglect alternative paths. These results highlight the importance of the level at which statistical data are represented, and suggest that when faced with hierarchical inference problems people adopt a simplifying heuristic that assumes alignment.


Animal models in vaccinology: state of the art and future perspectives for an animal-free approach.
Vaccine discovery and development is mainly driven by studies on immunogenicity and safety based on the appropriate animal models. In this review we will describe the importance of animal models in vaccinology, from research and development to pre-licensure and post-licensure commitments with particular emphasis on the advantages and limitations of each animal species. Finally, we will describe the most modern technologies, the new in vitro and ex vivo models and the new advances in the field which may drive into a new era of 'animal free' vaccinology.


Extraction Optimization of Mucilage from Seeds of Mimosa pudica by Response Surface Methodology.
Mimosa pudica seed mucilage (MPM) is composed of glucuronoxylan, which is a swellable, pH-responsive and non-toxic biomaterial. Herein, we aimed to extract MPM from M. pudica seeds (MP seeds) to ascertain optimization of extraction conditions to get highest yield by response surface methodology, via Box-Behnken design (RSM-BBD). MPM was extracted from MP seeds by a hot water extraction method. The effects of four different parameters on the extraction yield of MPM were evaluated: pH of the extraction medium (1-10), seed/water contact time (1-12 h), the temperature of extraction medium (30-90 °C), and seed/water ratio (1:5-1:35 w/v). The maximum yield of MPM obtained by Design-Expert software was 10.66% (10.66 g/100 g) at pH 7, seed/water contact time of 6 h, extraction temperature of 50 °C, and seed/water ratio of 1:20 w/v. The p values of ANOVA were found to be less than 0.0001, which indicated that the extraction yield of MPM was significantly affected by all the study parameters. The results revealed that pH and extraction temperature were the most significant factors affecting the yield of MPM. MPM in compressed tablet form showed pH-responsive on-off switching behavior at pH 7.4 and 1.2 in a reversible manner. MPM in compressed tablet form sustained the release of itopride for 16 h following a super case-II transport mechanism and zero-order release kinetics.


The Impact of Severe Acute Respiratory Syndrome-Coronavirus-2 Infection and Pandemic on Mental Health and Brain Function in the Elderly.
This review discusses the evolving evidence base and clinical considerations for examining the direct and indirect effects of the coronavirus disease (COVID-19) pandemic on the mental health of elderly individuals. It briefly addresses the cognitive and psychiatric outcomes in older adults who have survived COVID-19 infections and the complexity of appraising them during different stages of the pandemic. Indirect effects of the COVID-19 pandemic on the mental health of the geriatric population are also explored, including those influenced by quarantine, media campaigns, discrimination, and difficulties in accessing supportive services like long-term care and medical care.


The TelN/tos-assisted precise targeting of chromosome segments (TAPE).
Performing genomic large segmentation experiments will promote the annotation of complex genomic functions and contribute to the synthesis of designed genomes. It is challenging to obtain and manipulate large or complex DNA sequences with high efficiency. This study aims to develop an effective method for direct cloning of target genome sequences from different species. The TelN/tos system and a linear plasmid vector were first used to directly clone the large genomic segments in E. coli. For the in vitro cloning reaction, two telomeric sites were developed using TelN protelomerase at the end of the linear plasmid vector. The target DNA sequence can be easily hooked with the homology arms and maintained as a linear artificial chromosome with arbitrary restriction sites in a specific E. coli strain. Using the linear cloning strategy, we successfully cloned the bacterial DNA fragment of 156 kb, a yeast genomic fragment of 124 kb and mammalian mitochondrial fragment of 16 kb. The results showed a considerable improvement in cloning efficiency and demonstrated the important role of vector ratio in the cloning process. Due to the high efficiency and stability, TAPE is an effective technique for DNA cloning and fundamental molecular biotechnology method in synthetic biology.


Identifying chromatin features that regulate gene expression distribution.
Gene expression variability, differences in the number of mRNA per cell across a population of cells, is ubiquitous across diverse organisms with broad impacts on cellular phenotypes. The role of chromatin in regulating average gene expression has been extensively studied. However, what aspects of the chromatin contribute to gene expression variability is still underexplored. Here we addressed this problem by leveraging chromatin diversity and using a systematic investigation of randomly integrated expression reporters to identify what aspects of chromatin microenvironment contribute to gene expression variability. Using DNA barcoding and split-pool decoding, we created a large library of isogenic reporter clones and identified reporter integration sites in a massive and parallel manner. By mapping our measurements of reporter expression at different genomic loci with multiple epigenetic profiles including the enrichment of transcription factors and the distance to different chromatin states, we identified new factors that impact the regulation of gene expression distributions.


The nucleus tractus solitarii of the cat: a comparison of Golgi impregnated neurons with methionine-enkephalin- and substance P-immunoreactive neurons.
Golgi-impregnated and methionine-enkephalin (ME)- and substance P (SP)-immunoreactive neurons were studied throughout the feline nucleus tractus solitarii. The majority of Golgi-impregnated neurons in the NTS range in size from 5 to 18 micron. A noticeable exception is the large (15-30 micron) neurons of the ventrolateral subdivision. The Golgi-impregnated neurons possess dendritic trees which remain within the nucleus and even at times within the particular subdivisions. Golgi-impregnated neurons had a variety of spine forms: pedunculated, sessile, filiform, and complex. A number of neurons exhibited axons originating from the cell and they could be followed for distances up to 100 micron. ME- and SP-immunoreactive neurons were found in commissural, medial, lateral, and parvocellular subdivisions while ME-immunoreactive neurons were situated additionally in the intermediate and ventrolateral subdivisions. Both types of immunostained neurons were similar in size (6-20 micron) and shape of dendritic arbor. One population of ME-immunoreactive neurons resembled the large ventrolateral neurons of the Golgi impregnations. Neither type of immunostained neuron possessed the extensive dendritic arbor, numbers of spines, or axons of the Golgi-impregnated neurons. The presence of ME- and SP-immunoreactive neurons in regions which are associated with autonomic regulation suggests that these two peptides are involved in this process.


A non-randomized comparison of two radiotherapy protocols in inoperable squamous cell carcinoma of the oesophagus.
We report an audit of two different telebrachytherapy schedules in inoperable carcinoma of the oesophagus. Between October 1990 and December 1996, 108 patients with a Karnofsky performance status > or = 50 were selected from our database on the basis of intention to treat by telebrachytherapy. Teletherapy in the low dose group L (55 patients) consisted of 35 Gy in 15 fractions over 3 weeks, while that in the high dose group H (53 patients) consisted of 50 Gy in 25 fractions over 5 weeks. The choice of teletherapy dose was based on physician preference. The high dose rate intraluminal radiotherapy that followed 2 weeks later was identical in both groups and consisted of two applications of 6 Gy, a week apart. The pretreatment disease characteristics of the patients in both arms were similar. Relief of dysphagia was obtained in 49% of the patients in group L and in 75% of those in group H (chi2: P = 0.004). The median dysphagia-free interval was 0 and 7 months in groups L and H respectively (log-rank: P = 0.06), while the median overall survival was identical at 8 months (log-rank: P = 0.21) for both groups. The probability of survival at 1, 2 and 5 years was 34.8% versus 35.8%, 14.5% versus 13.9% and 0% versus 10% for groups L and H respectively. Morbidity in the form of ulcers, strictures and fistulae were observed in 9%, 7% and 5% of patients in group L compared with 8%, 8% and 13% in groups H respectively. This audit suggests that the protocol used in group H, when compared with group L, results in a greater proportion of patients being rendered dysphagia free, with a statistical trend towards a greater sustainment of dysphagia relief on follow-up.


Too Much or Too Little? How Much Control Should Patients Have Over EHR Data?
Electronic health records (EHRs) have been promoted as a mechanism to overcome the fragmented healthcare system in the United States. The challenge that is being discussed is the rights of the patient to control the access to their EHRs' data and the needs of healthcare professionals to know health data to make the best treatment decisions for their patients. The Federal Trade Commission has asked those who store consumer information to comply with the Fair Information Practice Principles. In the EHR context, these principles give the rights to the patient to control who can see their health data and what components of the data are restricted from view. Control is not limited to patients, as it also includes parents of adolescent children. We suggest that the ongoing policy discussion include consideration of the precise questions patients will be asked when a need for data sharing arises. Further, patients should understand the relative risks that they face, and the degree to which their decisions will (or will not) significantly reduce the risk of a data breach. As various approaches are considered, it is important to address the relative resource requirements and the associated costs of each option.


On the electrotransfer of polypeptides from gels to nitrocellulose membranes.
The conditions which affect the elution of polypeptides from polyacrylamide gels by electrophoresis and polypeptide-nitrocellulose interactions have been studied. The rate of elution of polypeptides from a 15% sodium dodecyl sulfate-polyacrylamide gel is dependent on the molecular weight of the individual polypeptides, which is in agreement with the results of W. N. Burnette (Anal. Biochem. 112, 195 (1981)). We also observed that current density affects the rate of elution. Polypeptides smaller than 20,000 daltons pass through pores of 0.45 microns, but not through the pores of 0.1-microns nitrocellulose membranes during electrophoresis. The nonionic detergent NP-40 inhibits the binding of polypeptides to nitrocellulose and removes prebound polypeptides from the membranes. Amido black and Coomassie blue staining and destaining processes do not remove the bound polypeptides from the membranes, but may affect the antigenicity of polypeptides. Polypeptides immobilized on nitrocellulose can be stored at -70 degrees C for future use.


Defense Mechanism of Phosphorothioated DNA under Peroxynitrite-Mediated Oxidative Stress.
DNA phosphorothioation (PT) exists in many pathogenic bacteria; however, the mechanism of PT-DNA resistance to the immune response is unclear. In this work, we meticulously investigated the peroxynitrite (PN) tolerance using PT-bioengineered E. coli strains. The in vivo experiment confirms that the S+ strain survives better than the S- strain under moderately oxidative stress. The LCMS, IC, and GCMS experiments demonstrated that phosphorothioate partially converted to phosphate, and the byproduct included sulfate and elemental sulfur. When O,O-diethyl thiophosphate ester (DETP) was used, the reaction rate k1 was determined to be 4.3 ± 0.5 M-1 s-1 in the first-order for both phosphorothioate and peroxynitrite at 35 °C and pH of 8.0. The IC50 values of phosphorothioate dinucleotides are dramatically increased by 400-700-fold compared to DETP. The SH/OH Yin-Yang mechanism rationalizes the in situ DNA self-defense against PN-mediated oxidative stress at the extra bioenergetic cost of DNA modification.


Successful transcatheter embolization of an unruptured idiopathic peripheral pulmonary artery aneurysm.
Pulmonary artery aneurysms (PAAs) occurring in the distal branch of the pulmonary artery are rare, and there are only a few reports of transcatheter embolization of unruptured idiopathic peripheral PAAs. We report a case of a 66-year-old woman with a history of surgery for pulmonary arteriovenous malformations who presented with an idiopathic peripheral PAA of A10 confirmed by contrast-enhanced computed tomography and pulmonary artery angiography. Transcatheter embolization was performed to prevent rupture, and the aneurysm was successfully embolized. Although there is no consensus on the indications and treatment for unruptured idiopathic peripheral PAAs, transcatheter embolization may be a favorable treatment option.


Aqueous Cr(VI) removal by a novel ball milled Fe0-biochar composite: Role of biochar electron transfer capacity under high pyrolysis temperature.
A novel ball milled Fe0-biochar composite was synthesized by ball milling the mixture of biochar (pyrolyzed at 300 °C, 500 °C, and 700 °C) and micron grade iron powder. FTIR, SEM, TEM-EDS, XRD, and XPS were applied to characterize this composite. XRD results showed that iron carbide phase was formed during the ball milling process. The ability of this synthesized composited to remove aqueous Cr(VI) was tested. Removal rates of Cr(VI) (49.6%, 65.8%, and 97.8%, respectively) by ball milled Fe0-biochar composite consisting of biochar pyrolyzed at 300 °C (300BMFe0-BC), 500 °C (500BMFe0-BC), and 700 °C (700BMFe0-BC) were much higher than those (19%, 11%, and 4%, respectively) by pristine biochar pyrolyzed at 300 °C (300BC), 500 °C (500BC), and 700 °C (700BC). Cr(VI) removal rate by 700BMFe0-BC increased from 15.4% to 97.8% when prolonging ball milling time from 6 h to 48 h. Ball milling promoted the combination of Fe0 and biochar as well as reduced the hydrodynamic diameter of the composite. Acidic conditions favored Cr(VI) removal. Ball milling exposed the functional groups of biochar and improved its Cr(VI) removal rate. Raman spectra showed that the degree of graphitization in 700 °C ball milled biochar (700BMBC) was the highest. Electrochemical analysis demonstrated that 700BMBC had the highest electron transfer capacity. In the presence of Fe0, graphitized structure in 700BMBC acted as an electron conductor, facilitating electron transfer from Fe0 to Cr(VI). Ball milling also destroyed the surface iron oxide layer to regenerate the composite.


Hypofractionated Proton Beam Therapy for cT1-2N0M0 Non-small Cell Lung Cancer Patients With Interstitial Lung Disease.
To evaluate the outcomes of proton beam therapy (PBT) for early-stage non-small cell lung cancer (NSCLC) in patients with interstitial lung disease (ILD). Between 2002 and 2017, 110 patients receiving hypofractionated PBT for cT1-2N0M0 NSCLC were reviewed. Of the 110 patients, 17 were diagnosed with ILD. The median follow-up period was 37.8 months. No significant difference in the 1-year cumulative rate of grade ≥2 pneumonitis was observed between patients with and those without ILD (17.6% vs. 14.1%, p=0.708). The lung doses were significantly lower in patients with than in those without ILD among patients without grade ≥2 pneumonitis. There were no significant differences in overall survival or local recurrence-free rates according to the presence of ILD. PBT appears to be a feasible and effective treatment for cT1-2N0M0 NSCLC in patients with ILD, but the lung dose should be strictly reduced.


DNA methylation changes in cervical cancers.
Cervical carcinoma is one of the major causes of death in women worldwide. It is difficult to foresee a dramatic increase in cure rate even with the most optimal combination of cytotoxic drugs, surgery, and radiation; therefore, testing of molecular targeted therapies against this malignancy is highly desirable. Cervical cancer is a multistep process with accumulation of genetic and epigenetic alterations in regulatory genes, leading to activation of oncogenes and inactivation or loss of tumor suppressor genes (TSGs). In the last decade, in addition to genetic alterations, epigenetic inactivation of TSGs by promoter hypermethylation has been recognized as an important and alternative mechanism in tumorigenesis. In cervical cancer, epigenetic alterations can affect the expression of papillomavirus as well as host genes in relation to stages representing the multistep process of carcinogenesis. Here we discuss these epigenetic alterations in cervical cancer focusing on DNA methylation.


Mass spectrometry cancer data classification using wavelets and genetic algorithm.
This paper introduces a hybrid feature extraction method applied to mass spectrometry (MS) data for cancer classification. Haar wavelets are employed to transform MS data into orthogonal wavelet coefficients. The most prominent discriminant wavelets are then selected by genetic algorithm (GA) to form feature sets. The combination of wavelets and GA yields highly distinct feature sets that serve as inputs to classification algorithms. Experimental results show the robustness and significant dominance of the wavelet-GA against competitive methods. The proposed method therefore can be applied to cancer classification models that are useful as real clinical decision support systems for medical practitioners.


A comprehensive review on clinically proven natural products in the management of nerve pain, with mechanistic insights.
People are treating their neuropathic pain with several approved and licensed pharmacological drugs. But due to having existing limitations like low efficacy with some side effects, there needs to be a more effective alternative and complementary therapeutic options. s: The study was designed to discuss the mechanistic role of several clinically proven natural products that have been shown to play a significant role against different nerve pain or neuropathic pain. ology: Information for this review article was salvaged using several accessible searching databases like SciVerse Scopus ® (Elsevier Properties S. A, USA), Web of Science® (Thomson Reuters, USA), and PubMed® (U.S. National Library of Medicine, USA) considering some search items like - nerve pain, natural products in pain/nerve pain management, clinically proven natural products in pain management, pain-reducing agents and so on. Our study reported the therapeutic efficacy of natural products and their possible mechanism against neuropathic pain in the human body. Natural products widely used to treat neuropathic pain include comfrey root extract ointment, lavender oil, Rose Oil, aromatic essential oil, ginger oil, vitex agnus-castus, peganum oil, and ajwain 10%. Some common pathways are involved in pain relief through sensory stimulation, enzymatic, anti-inflammatory, and pain-related receptor regulation. The present study suggests that the mentioned natural products can be an appropriate choice for the treatment and management of neuropathic pain.


Can the laryngeal mask play a role in electroconvulsive treatment? A pilot study.
Optimising ventilation control in the anaesthetic process of electroconvulsive therapy (ECT) can be a method for improving the effectiveness and safety of some treatments. There are several approaches for inducing adequate therapeutic seizures in patients with medical co-morbidity or in those with high seizure thresholds, although all of them present limitations. A new approach is hyperventilation with laryngeal mask, a method that improves oxygenation, achieves hypocapnia, and lowers seizure threshold. Thus, hyperventilation with laryngeal mask could, hypothetically, be useful to decrease the energy needed to obtain adequate therapeutic seizures. A randomized crossover study was conducted on 14 patients on maintenance ECT, using a laryngeal mask versus a facemask. When laryngeal masks were applied, the energy was reduced by 45% compared with the energy required when ventilated with facemasks (performed with standard dose). The results of the study revealed that, despite a 45% energy reduction with laryngeal mask, there were no significant differences in seizure length in either group. These results prompt us to propose ventilation with laryngeal mask as an alternative to decrease energy applied without lowering seizure times. This is a preliminary study with a small sample, which opens new lines of research. Larger samples and other measurements of seizure adequacy are needed in future studies, in order to study possible improvements in ventilation and minimising adverse effects of ECT, especially in patients with medical co-morbidities.


Viral Infection at High Magnification: 3D Electron Microscopy Methods to Analyze the Architecture of Infected Cells.
As obligate intracellular parasites, viruses need to hijack their cellular hosts and reprogram their machineries in order to replicate their genomes and produce new virions. For the direct visualization of the different steps of a viral life cycle (attachment, entry, replication, assembly and egress) electron microscopy (EM) methods are extremely helpful. While conventional EM has given important information about virus-host cell interactions, the development of three-dimensional EM (3D-EM) approaches provides unprecedented insights into how viruses remodel the intracellular architecture of the host cell. During the last years several 3D-EM methods have been developed. Here we will provide a description of the main approaches and examples of innovative applications.


[Traumatic Lacrimal Duct Stenosis: Demographics and Success Rate of Surgical Procedures for Secondary Treatment].
Traumatic lacrimal duct stenosis can be the result of sharp trauma of the eyelid, indirect trauma or surgery in the nasoorbital region, as well as burns, chemotherapy and radiation of the facial region. The aim of the study is to present the demographics, patient satisfaction, and course of different surgical procedures for secondary treatment of traumatic lacrimal duct stenosis. We retrospectively reviewed the medical records of 50 patients who required surgery for traumatic lacrimal duct stenosis from 2009 to 2011 at the University Eye Hospital in Muenster. The evaluation included the following criteria: age, sex, duration of symptoms, complication rate and the rate of recurrence. We systematically evaluated the medical data and asked the patients about the recent symptoms via telephone questionnaire. 60 eyes of 50 patients were included (18 women; 32 men; age: 51.8 ± 17.1 years); 34 eyes (56.7 %) had already had operations of the lacrimal system. The success rate was 58.1 %. The mean postoperative follow-up was 52.3 ± 10.7 months. In 11 cases (18.3 %), a dakryocystorhinostomy (DCR) was necessary. Traumatic lacrimal stenosis is more common in men, and has a poorer prognosis than lacrimal duct stenosis from other causes.


Home Health Agency Performance in the United States: 2011-15.
To evaluate home health agency quality performance. Observational study. Home health agencies. All Medicare-certified agencies with at least 6 months of data from 2011 to 2015. Twenty-two quality indicators, five patient survey indicators, and their composite scores. The study included 11,462 Medicare-certified home health agencies that served 92.4% of all ZIP codes nationwide, accounting for 315.2 million people. The mean composite scores were 409.1 ± 22.7 out of 500 with the patient survey indicators and 492.3 ± 21.7 out of 600 without the patient survey indicators. Home health agency performance on 27 quality indicators varied, with the coefficients of dispersion ranging from 4.9 to 62.8. Categorization of agencies into performance quartiles revealed that 3,179 (27.7%) were in the low-performing group (below 25th percentile) at least one time during the period from 2011-12 to 2014-15 and that 493 were in the low-performing group throughout the study period. Geographic variation in agency performance was observed. Agencies with longer Medicare-certified years were more likely to have high-performing scores; agencies providing partial services, with proprietary ownership, and those with long travel distances to reach patients had lower performance. Agencies serving low-income counties and counties with lower proportions of women and senior residences and greater proportions of Hispanic residents were more likely to attain lower performance scores. Home health agency performance on several quality indicators varied, and many agencies were persistently in the lowest quartile of performance. Still, there is a need to improve the quality of care of all agencies. Many parts of the United States, particularly lower-income areas and areas with more Hispanic residents, are more likely to receive lower quality home health care.


Calculated infrared spectra of nerve agents and simulants.
Since organophosphorus nerve agents are among the most toxic known chemical warfare agents, it is desirable to have a way to distinguish between one and another. Infrared spectroscopy is a common tool for identifying molecules. Given the difficulty in handling these chemicals, calculated IR spectra can be useful. Calculated IR spectra are presented for G agents, V agents, and simulants. Quantum chemistry calculations were performed using the Gaussian 03 package at the B3LYP/6-31+G(d,p) level of theory. The most prominent IR lines are due to vibrations of P-O-C and P=O groups within the molecules. It should be possible to distinguish between the G-series and V-series agents using IR spectroscopy, but unique identification of individual chemical agents is unlikely.


Expressions of ER, PR, HER-2, COX-2, and VEGF in primary and relapsed/metastatic breast cancers.
In the present study, we evaluated expressions of estrogen receptor (ER), progestin receptor (PR), human epidermal growth factor receptor-2 (HER-2), cyclooxygenase-2 (COX-2), and vascular endothelial growth factor (VEGF) in primary and relapsed/metastatic breast cancers to elucidate the clinical significance of these markers. The markers were evaluated by immunohistochemistry in specimens of 50 patients with primary or metastatic breast cancer. Positive rates of ER were significantly (p = 0.002) higher in primary versus relapsed/metastatic breast cancer (70 vs. 38 %, respectively). The VEGF positive expression rates were also significantly higher in primary versus metastatic cancer (82 vs. 38 %, respectively; p < 0.001). By contrast, positive rates of HER-2 and COX-2 were not significantly different between different types of cancer. COX-2 correlated with HER-2 expression in both primary and relapsed/metastatic focuses of breast cancer. COX-2 also correlated with VEGF expression in primary breast cancer. Expressions of ER, PR, HER2, and COX-2 did not correlate between primary and relapsed/metastatic breast cancers, indicating that the treatment decision should be made according to the status of these markers in relapsed/metastatic focuses. The total change rates of ER, PR, HER-2, COX-2, and VEGF were 26, 18, 10, 30, and 58 %, respectively. In conclusion, HER-2 and COX-2, along with VEGF, appear to play a role in the development and progression of breast cancer. In addition, all of the studied markers may serve as indicators of prognosis.


Cyclosporin partitions into phospholipid vesicles and disrupts membrane architecture.
Cyclosporin is a fungal metabolite demonstrating potent immunosuppressive activity both in vitro and in vivo, but the mechanism of action is poorly understood. Using [3H]dihydrocyclosporin C ([3H]CsC) we observed significant binding by mononuclear cells, erythrocytes and phosphatidyl choline (PC) vesicles which was reversible by the addition of excess CsA. Trypsin, pronase or heat treatments demonstrated that B cells and adherent cells express a protease-sensitive membrane binding site not observed on T cells. The nature of the interaction between CsA and the PC vesicles was studied using the membrane surface probe 1-anilino-8-naphthyl sulfonic acid (ANS-). ANS- -induced fluorescence was reduced by 24% in the presence of 4.75 X 10(-7). M CsA indicating that CsA displaces ANS- from the PC vesicles. CsA also effected a shift in the phase transition temperature of PC vesicles from 23 degrees C to 19 degrees C. Finally, the rate of concanavalin A (Con A)-induced cap formation by T lymphocytes was approximately doubled in the presence of 2.6 X 10(-5) M CsA. These data demonstrate that CsA partitions into phospholipid vesicle membranes and the plasmalemma of mononuclear cells resulting in an increased membrane fluidity.


Long-term outcomes for children with an incidentally discovered Chiari malformation type 1: what is the clinical significance?
Chiari malformation type 1 (CM1) is an increasingly common incidental finding on magnetic resonance imaging (MRI). The proportion of children with an incidentally discovered CM1 who upon further evaluation require operative intervention for previously unrecognized signs and symptoms of neurological compromise or significant radiographic findings (syringomyelia) is unclear. An extensive long-term single-institution patient series was evaluated to better clarify the likelihood of surgery in patients who present with an incidentally discovered CM1. This study was conducted using prospective data for patients up to 18 years old that were evaluated for a CM1 at a large tertiary pediatric neurosurgery clinic between February 2009 and June 2019. Patients were excluded if they did not have an incidentally discovered CM1 and at least 12 months of clinical follow-up. A total of 218 consecutive patients were included in this study. The mean age at the initial neurosurgical evaluation was 6.5 years (range 5 months to 18.4 years), and the mean duration of clinical follow-up was 40.6 months (range 12 to 114 months). Initial MR imaging was most commonly obtained for the evaluation of seizures (15.1%), nonspecific headaches (not occipital or tussive) (14.7%), trauma (9.6%), and developmental delay (7.8%). Of the patients studied that eventually required surgery, we identified two groups: those operated before 6 months since presentation and those operated after 6 months. A total of 36 patients (16.5%) underwent a decompression with 22 patients (61.1%) receiving surgery within 6 months and the remaining 14 patients (38.9%) beyond 6 months. Patients undergoing early surgery (10.1%) initially presented with a significant syrinx or were noted to have an occult neurological dysfunction, whereas a smaller subset of patients (6.4%) eventually required surgery over time due to the development of new symptoms or a de novo syrinx. Only the presence of syringomyelia was statistically significant for the need of a surgical intervention, while age, sex and degree of tonsillar herniation were not. Evaluation of a large group of patients with an incidentally discovered Chiari malformation demonstrated that most patients may be managed conservatively, especially in the absence of syringomyelia. However, there is a subset of patients who will go on to develop a de novo syrinx or neurological symptoms that are new or progressive during follow-up, which should be evaluated by imaging of the brain and spinal cord. The presence of syringomyelia was associated with need for early surgical intervention. However, for patients without syringomyelia, surgical intervention is uncommon but may be delayed up to several years after presentation; therefore, long-term clinical follow-up is recommended.


Trigger for intercellular adhesion molecule-1 expression in rat lungs transplanted from non-heart-beating donors.
Lung transplantation from non-heart-beating donors causes ischemia-reperfusion injury. We sought to determine the trigger for expression of intercellular adhesion molecule-1 (ICAM-1) caused by ischemia-reperfusion injury. Thirty-six Sprague-Dawley rats underwent left lung transplant (six groups of 6). Lungs were transplanted immediately after arrest, or from non-heart-beating donors after 2 hours of oxygen-ventilation or no ventilation. Recipients were reperfused for 4 or 6 hours, then lungs were stained with a mouse anti-rat ICAM-1 monoclonal antibody, developed with avidin-biotin peroxidase to a biotinylated anti-mouse immunoglobin G antibody. Intercellular adhesion molecule-1 expression was graded by two masked observers as 0 = absent, 1 = weak, or 2 = strong in alveoli, arterioles, and venules. Explanted recipient left lungs served as negative controls, and positive controls were generated 6 hours after intraperitoneal injection of endotoxin. Intercellular adhesion molecule-1 expression above baseline among groups was compared by Fisher's exact test. Constitutive expression of ICAM-1 was present in rat lung alveoli, with 24 of 35 controls staining weakly and 4 of 35 strongly positive in alveolar areas. Intercellular adhesion molecule-1 expression was not increased in transplanted lungs evaluated after 4 hours of reperfusion, even lungs retrieved from non-heart-beating donors. But when non-heart-beating donor lungs were assessed 6 hours after onset of reperfusion, ICAM-1 expression was significantly more apparent in alveolar and arteriolar areas, compared with controls and lungs transplanted immediately after arrest. Lungs transplanted immediately after circulatory arrest do not sustain sufficient ischemia-reperfusion injury to upregulate ICAM-1. Onset of reperfusion is the signal for ICAM-1 expression, not the onset of ischemia or the total duration of ischemic and reperfusion time together. Strategies at reperfusion may minimize ICAM-1 expression.


Mitochondrial gene expression is regulated at multiple levels and differentially in the heart and liver by thyroid hormones.
Biogenesis of the oxidative phosphorylation system (OXPHOS) requires the coordinated expression of the nuclear and the mitochondrial genomes. Thyroid hormones play an important role in cell growth and differentiation and are one of the main effectors in mitochondrial biogenesis. To determine how mtDNA expression is regulated, we have investigated the response of two different tissues, the heart and liver, to the thyroid hormone status in vivo and in vitro. We show here that mtDNA expression is a tightly regulated process and that several levels of control can take place simultaneously. In addition, we show that the mechanisms operating in the control of mtDNA expression and their relevance differ between the two tissues, being gene dosage important only in heart while transcription rate and translation efficiency have more weight in liver cells. Another interesting difference is the lack of a direct effect of thyroid hormones on heart mitochondrial transcription.


Relative efficacies of insulin and poly (lactic-co-glycolic) acid encapsulated nano-insulin in modulating certain significant biomarkers in arsenic intoxicated L6 cells.
The present study evaluates relative efficacies of insulin and PLGA-loaded-nano-insulin (NIn) in combating arsenic-induced impairment of glucose uptake, insulin resistance and mitochondrial dysfunction in L6 skeletal muscle cells. L6 cells were treated with 0.5 mM sodium arsenite for 30 min and then the cells were further treated with either 100 nM insulin (standardized dose) or either of the two doses, 50 nM and 100 nM of nano-insulin. Various biomarkers like pyruvate-kinase and glucokinase, ATP/ADP ratio, mitochondrial membrane potential, cytosolic release of mitochondrial cytochrome c, cell membrane potential and calcium-ion level were studied and analyzed to ascertain the status of mitochondrial functioning in all experimental and control sets of L6 cells. The size, morphology and zeta potential of formulated NIn were determined by using dynamic light scattering, scanning electronic and atomic-force microscopies. Expression of signalling cascades like GLUT4, IRS1, IRS2, UCP2, PI3, and p38 was critically analyzed. Overall results suggested that both insulin and NIn improved mitochondrial functioning in arsenite-intoxicated L6 cells, NIn showing better effects at a much lower dose (at nearly 10-fold decreased dose) than that produced by insulin. Nano-insulin being non-toxic and effective at a much lesser dose, therefore, has potential for therapeutic use in the management of arsenic induced diabetes.


Transport of circulating serum cholesterol by human renal cell carcinoma.
Clear cell renal cancer contains a large quantity of cholesterol ester (300-mg./gm. protein). To determine whether abnormalities in cholesterol transport could account for this sterol accumulation, the uptake, release, and imaging capabilities of intravenously injected 131I-6-iodomethyl-29-norcholesterol, a cholesterol analogue, were studied preoperatively in five patients with clear cell renal cancer. At surgery, samples of the liver, tumor, adrenal, and non-tumor kidney were obtained for analysis. 131I-sterol uptake by the tumor, when normalized for cholesterol content, was less than for adrenal, liver or kidney. In contrast, release of preloaded 131I-sterol from the human tumors was consistently slower than for normal kidney. The reduced release of free cholesterol from renal cancer cells may, in part, be responsible for the accumulation of cholesterol in human renal cancer.


Structure and function of the bc-complex of Rhodobacter sphaeroides.
The ubiquinol:cytochrome c2 oxidoreductase (bc-complex) of Rhodobacter sphaeroides has three main subunits, which bear the prosthetic groups, and contribute to three catalytic sites and internal electron transfer pathways which define the modified Q-cycle mechanism. In this paper, we report on progress in modelling the structure of the bc-complex, and experiments using site directed mutagenesis and biophysical assay to probe the structural and function consequences of specific modifications to these subunits.


IgE-mediated 15-hydroxyeicosatetraenoic acid (15-HETE) generation by peripheral blood leukocytes: its association with basophil activation.
Allergen-induced basophil activation has been associated with the release of several mediators and with an increased expression of CD203c molecules on basophils. To assess the influence of specific allergens on the generation of 15-hydroxyeicosatetraenoic (15-HETE) from peripheral blood leukocytes in relation to basophil activation, on the basis of CD203c molecule expression and histamine release. The study included 15 patients with clinical symptoms of birch pollen allergy confirmed by a positive skin prick test with the birch allergen, and 6 healthy controls. Leukocytes isolated from peripheral blood were incubated with 3 concentrations of the birch pollen allergen (Bet v 1), anti-IgE or with ionophore A23187. In vitro challenge of leukocytes from allergic patients with 1 ng/ml of allergen induced a significant increase in 15-HETE generation. An increase above 30% was observed in almost half the allergic patients, with mean values ranging from 40% to 46%, but not in healthy controls. Anti-IgE antibodies increased 15-HETE generation in 5 patients (termed IgE+), and the allergen induced a significant increase in 15-HETE in all patients who reacted to anti-IgE. The mean CD203c expression on basophils of the allergic patients increased after allergen challenge, but a significant increase (> 30%) was observed only in patients who demonstrated an increased expression after anti-IgE exposure. A significant correlation was seen between 15-HETE generation and histamine release induced by the highest concentration of the allergen (r = 0.95; p < 0.01). Allergen-induced, IgE-mediated activation of basophils is associated with a significant increase in 15-HETE generation.


Isolation and biological characteristics of sheep amniotic epithelial cells.
Amniotic epithelial cells (AECs), isolated from placenta, have epithelial cells and stem cells characteristics. Most of the previous studies focused on the biological characteristics of human amniotic epithelial cells, which demonstrated amniotic epithelial cells not only had low immunogenicity and potent potential to differentiate into three germ layers, but also could secrete various immunomodulatory factors. However, compared to study on human amniotic epithelial cells, few studies have been done on other animals. In this study, sheep amniotic epithelial cells were successfully isolated and their surface makers were accessed by immunofluorescence assay, and found that AECs were expressed Oct4 and Sox2, which were necessary for maintaining the undifferentiated state of pluripotent stem cells. Based on cloning efficiency and growth kinetics assay, AECs were found to possess self-renewal capacity and the growth curve was S-shaped. In addition, AECs could be induced into adipocytes, osteoblasts and chondrocytes in vitro, showing they had multi-differential ability. Reverse transcription-polymerase chain reaction results showed that AECs expressed CD29, CD44, CD90 and CK19, and didn't expressed CD34, CD45 and the telomerase gene (TERT). Little change in chromosome number was observed in AEC cultures for up to at least the first ten passages. In summary, this study results revealed that sheep AECs possessed more advantages for cell therapy and might play a key role in cell therapy and regenerative medicine in the future.


Postoperative application of amphotericin B nasal spray in chronic rhinosinusitis with nasal polyposis, with a review of the antifungal therapy.
Chronic rhinosinusitis (CRS) affects 1-4% of the adult population. The etiology of this multifactorial, chronic disease, which leads to a significant impairment of the quality of life, often accompanied by nasal polyposis, is not fully understood. In the past decade, it was presumed that the disease, which causes characteristic eosinophilic infiltration of the nasal mucosa, is triggered by an enhanced (but not classical allergic IgE-type) immune response against fungal organisms in the nasal mucus. If this supposition is correct, then it appears obvious that the administration of amphotericin B nasal spray in adequate concentration following endoscopic polypectomy should be advantageous for these patients, and might even reduce the number of recurrent cases. To check on this assumption, we conducted a prospective randomized placebo-controlled trial involving 33 patients, 30 of whom remained in the study throughout. Patients with nasal polyposis were operated on with an endoscopic technique between 1 November 2005 and 1 October 2006; group A (14 randomly selected patients) were treated with a nasal spray containing 5 mg/ml amphotericin B, while the placebo group B (16 randomly selected patients) received a nasal spray lacking amphotericin B. We evaluated our results with the aid of a modified Lund-Mackay CT score, the SNAQ-11 test (which assesses changes in the symptoms), a quality of life test and endoscopy. The SPSS 14.0 for Windows program was utilized to process the data of examinations performed preoperatively and 1 year postoperatively. The CT scores of the group A patients 1 year after the operation exhibited wide scattering, without signs of recovery. The CT scores of the group B patients indicated a slight improvement, though this did not prove significant relative to group A. Both the SNAQ-11 test and the quality of life test revealed a significant improvement in each group, but the degrees of change in these tests did not differ significantly between the two groups of patients. The endoscopic findings indicated a slight improvement to the advantage of the amphotericin B-treated group 12 months after the operation. These results lead to the conclusion that the administration of amphotericin B nasal spray to patients operated on for nasal polyposis does not give rise to a significant alteration in either CT score, clinical symptoms, or quality of life. The more favorable clinical aspects observed in the amphotericin B-treated group during the endoscopic follow-up did not correspond to an improvement in the symptoms. In connection with the conclusions drawn from this study, the authors discuss the controversial data available on the fungal etiology of CRS. They critically analyze the contradictory observations and conclusions of seven recent clinical studies.


IgD+IgM- B cells mount immune responses that exhibit altered antibody repertoire.
IgM and IgD expression during B cell development and differentiation is strictly and developmentally controlled. Although studies have suggested subtle differences in B cell activation, tolerance, and affinity maturation when antigens ligate cell membrane IgM or IgD, the mechanisms that may explain these differences remain unknown and no drastic differences in immune responses have been reported in mice whose B cells selectively lack IgM or IgD. We now show that the antibody repertoire in IgM(-/-) mice is dramatically altered during the primary response to phosphorylcholine. In IgM(-/-) mice, B cells that are activated and differentiate into antibody-forming cells and germinal center B cells express VH genes other than the T15 genes that dominate in wild-type mice. The kinetics of the antigen-specific IgD primary antibody response in IgM(-/-) mice appears similar to that of IgG, but not to that of IgM in wild-type mice. Thus, our studies demonstrate that differences in the roles played by IgM and IgD in regulating the responsiveness and differentiation of B lymphocytes can have major biological consequences during adaptive immune responses.


Photoprotection of 1,4-dihydropyridine derivatives by dyes.
The possibility of increasing the photochemical stability of nisoldipine by using indigotine and azorubine as photoprotectors has been studied. The course of the photodegradation was monitored by means of UV-vis spectrophotometry and HPLC. Quantitative assessments of the nisoldipine photodegradation included evaluation of the quantum yields and kinetic parameters. In order to establish the light intensity absorbed by a system, Reinecke salt was used as a chemical actinometer. The values of the quantum yields (phi) of photodegradation decreased with increasing dye concentration and were 0.24-0.15 and 0.27-0.25 for indigotine and azorubine, respectively. Furthermore, our attention was focused on determination the role of the selected dyes during the photodegradation of nisoldipine and the calculations were made to eliminate an inner filter effect. The values obtained were used to construct a Stern-Volmer plot and calculate the Stern-Volmer constants (KSV). On the basis of the KSV and the values of the quenching constant (kq) the exited state lifetime (tauo) of nisoldipine in the presence of indigotine and azorubine were estimated. The calculated values of tauo for NS were 6.66x10(-6)s--in the presence of indigotine and 2.76x10(-6)s--in the presence of azorubine, indicating that the photodegradation of nisoldipine occurs from the lowest triplet excited state.


CT-based renal volume measurements: correlation with renal function in patients with renal tumours.
To evaluate the correlations between renal cortical volume (RCV), renal parenchymal volume (RPV), and renal function in patients with renal tumours before and after laparoscopic partial nephrectomy (LPN). Thirty-five patients with a single unilateral renal tumour who had undergone contrast-enhanced computed tomography (CT) and renal nuclear scintigraphy before and after LPN were retrospectively studied. RCV and RPV were calculated as renal volume, excluding tumours or cysts, using a semi-automatic segmentation program. The correlations between RCV, RPV, and glomerular filtration rate (GFR) were undertaken preoperatively and postoperatively using the Pearson correlation coefficient. Preoperatively, the correlations between RCV and GFR, and RPV and GFR for the operated kidneys was r=0.502 (p=0.002) and 0.527 (p=0.001), respectively, whereas the correlations for the contralateral side were r=0.384 (p=0.023) and r=0.412 (p=0.014). The mean RCV and RPV of the operated kidneys decreased by 27.4% and 24.8%. The mean split GFR of the operated kidneys decreased by 36.4%. Postoperatively, residual RCV (r=0.619, p<0.001) and RPV (r=0.593, p<0.001) correlated moderately with the GFR of the operated kidneys. Renal volume, both RCV and RPV, had a moderate relationship with renal function before and after operation. CT-based renal volume measurements could serve as a simple and effective method for estimation of postoperative renal function.


Micrometastases in sentinel nodes of gastric cancer.
The sentinel node (SN) is the first lymph node in the lymphatic basin to be affected by metastasis from the primary tumour and is used to predict the status of the remaining nodes in the basin. We succeeded in detecting SNs of clinically early gastric cancers by intraoperative injection of a blue dye around the tumour. In the study presented here, multiple-marker reverse transcription-polymerase chain reaction (RT-PCR) was used to detect micrometastases in SNs and results were compared with those obtained with conventional histology. Expressions of cytokeratin-18 (CK-18), carcinoembryonic antigen (CEA), human telomerase reverse transcriptase (hTRT) and MUC-1 in SNs were determined by RT-PCR and Southern blot assay. Of the 213 SNs obtained from 35 cases of gastric cancer, eight nodes (3.8%) from five patients contained metastases that could be identified by conventional histology. However, CK-18 mRNA was expressed in 15 (7.0%), CEA in 12 (5.6%), hTRT in 10 (4.7%), and MUC-1 in 12 (5.6%) nodes, with at least one mRNA marker expressed in 25 nodes (11.7%) obtained from six patients. In the five patients with nodal metastases identified by conventional histology, two had metastases in both SNs and non-SNs. And, in the 30 patients without nodal metastases identified by conventional histology, one patient with micrometastases in the SNs identified by RT - PCR and Southern blot assay also had metastases in non-SNs as identified by serial sectioning and immunostaining of CK-18. All additional metastases were detected in non-SNs located in the same lymphatic basin as the previously detected SNs. This suggests that lymph node dissection of early-stage gastric cancer in the lymphatic basin may be mandatory even for patients without histologically detectable metastases in SNs.


Developmental changes of heat shock protein 73 in human brain.
The expression of heat shock protein (HSP) 73, a constitutive form of HSP, was evaluated immunohistologically in human brains, from embryos to adults. HSP 73 immunoreactivity was first detected in the embryo at 6 weeks of gestational age (GW) in the ventral horn cells of the spinal cord and the dorsal root ganglion cells. During the fetal period, the reactivity extended cranially, becoming detectable in the cerebral pyramidal cells at 40 GW. Glial cells in the spinal cord also showed HSP 73 immunoreactivity, from 22 GW. The time course of the development of HSP 73 immunoreactivity was mostly consistent with the time courses of overall neuronal and glial maturation, suggesting an increasing role of HSP 73 during neural cell differentiation.


Using big data to retrospectively validate the COMPASS-CAT risk assessment model: considerations on methodology.
External validation is a prerequisite in order for a prediction model to be introduced into clinical practice. Nonetheless, methodologically intact external validation studies are a scarce finding. Utilization of big datasets can help overcome several causes of methodological failure. However, transparent reporting is needed to standardize the methods, assess the risk of bias and synthesize multiple validation studies in order to infer model generalizability. We describe the methodological challenges faced when using multiple big datasets to perform the first retrospective external validation study of the Prospective Comparison of Methods for thromboembolic risk assessment with clinical Perceptions and AwareneSS in real life patients-Cancer Associated Thrombosis (COMPASS-CAT) Risk Assessment Model for predicting venous thromboembolism in patients with cancer. The challenges included choosing the starting point, defining time sensitive variables that serve both as risk factors and outcome variables and using non-research oriented databases to form validated definitions from administrative codes. We also present the structured plan we used so as to overcome those obstacles and reduce bias with the target of producing an external validation study that successfully complies with prediction model reporting guidelines.


Protein S and protein C level changes with adjuvant tamoxifen therapy in postmenopausal women.
Tamoxifen citrate is a synthetic antiestrogen that provides survival benefit when given as adjuvant treatment in postmenopausal women with breast cancer. Venous thrombophlebitis may complicate tamoxifen treatment at a rate of approximately one per 800 treatment-years. To explore the possible procoagulant effects associated with tamoxifen therapy we evaluated changes in protein S and C activity levels in 58 postmenopausal women with surgically resected breast cancer who were disease-free and participating in a double-blind, placebo-controlled, randomized toxicity study of tamoxifen. The changes in protein C activities for the tamoxifen group (mean level = 113%) compared to those in the placebo group (mean level = 115%) were not significant (p = 0.45). Protein S activity levels increased while protein C activity levels decreased from baseline at 24 months in both tamoxifen and placebo groups. We conclude that the possible thrombophlebitis-promoting effect of tamoxifen in postmenopausal women is unlikely to be explained on the basis of protein S and protein C activity level changes.


Potential Role of Natural Polyphenols against Protein Aggregation Toxicity: In Vitro, In Vivo, and Clinical Studies.
One of the main features of neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease is the amyloidogenic behavior of disease-specific proteins including amyloid β, tau, α-synuclein, and mutant Huntingtin which participate in the formation, accumulation, and deposition of toxic misfolded aggregates. Consequently, these proteins not only associated with the progress of their respective neurodegenerative pathologies but also qualify as disease-specific biomarkers. The aim of using natural polyphenols is to target amyloid-dependent proteopathies by decreasing free radical damage and inhibiting and dissolving amyloid fibrils. We explore the effectiveness of the polyphenols epigallocatechin-3-gallate, oleuropein aglycone, and quercetin on their ability to inhibit aggregation of amyloid β, tau, and α-synuclein and mitigate other pathological features for Alzheimer's disease and Parkinson's disease. The analysis was carried from in vitro and cell line studies to animal models and clinical trials. This Review describes the use of phytochemical compounds as prophylactic agents for Alzheimer's disease, Parkinson's disease, and other proteopathies.


Inflammation-triggered local drug release ameliorates colitis by inhibiting dendritic cell migration and Th1/Th17 differentiation.
Enteric-coated formulations using Eudragit® polymers have been extensively used for delivering drugs to the lower gastrointestinal tract. However, these drug-delivery systems cannot accurately deliver the therapeutic cargoes to colon because of early degradation of the polymers at alkaline pH of the small intestine. Here, we describe a precise method of delivering drugs to inflammation sites in colon using an oral drug delivery system. Tacrolimus (FK506)-loaded microspheres were prepared using a thioketal-based polymer that releases drug in response to reactive oxygen species (ROS), which are abundantly produced at the sites of inflammation in acute colitis. Orally-administered FK506-loaded thioketal microspheres (FK506-TKM) led to a substantial accumulation of FK506 in inflamed colon and effectively alleviated dextran-sulfate sodium (DSS)-induced murine colitis. At the molecular level, FK506-TKM significantly inhibited infiltration of CD4+ and CD8+ T lymphocytes in colon and differentiation of CD4+ T cells into Th1 and Th17 cells in colon-draining mesenteric lymph nodes via restricting dendritic cell migration from colon. Our findings indicate orally-administered thioketal-based drug delivery system as a promising means of treating acute inflammatory bowel diseases.


Aspergillus oryzae atfB encodes a transcription factor required for stress tolerance in conidia.
Using an Aspergillus oryzae EST database, we identified a gene encoding a transcription factor (atfB), which is a member of the ATF/CREB family. Expression of atfB was barely detectable during vegetative growth, but was readily detected during conidiation in solid-state culture. Microarray analyses showed that expression of many other genes, including catalase (catA), were downregulated in an atfB-disruptant. The expression of most of these genes was upregulated in the wild-type strain during the conidiation phase in solid-state culture, and the expression pattern was similar to that of atfB itself. In the absence of stress, e.g. heat-shock or hydrogen peroxide, the conidial germination ratios for the DeltaatfB strain and the wild-type strain were similar, but the stress tolerance of conidia carrying the DeltaatfB deletion was less than that of the wild-type conidia. CRE-like DNA motifs, which are bound by ATF/CREB proteins, were found in the promoters of most of the downregulated genes in the DeltaatfB strain. Thus, atfB appears to encode a transcription factor required for stress tolerance in conidia.


High dose cyclophosphamide as an adjunct to radiotherapy for advanced seminoma.
Two patients with far-advanced seminoma of the testis were given single intravenous injections of high dose (1.5 and 2.0 gm. per m.) cyclophosphamide. Both patients experienced prompt, dramatic objective regressions without severe toxicity. Definitive radiotherapy, deemed technically unfeasible before cyclophosphamide administration, was administered subsequently. Long-term complete remissions (more than 16 and 28 months) have been attained in both patients. High dose cyclophosphamide may prove useful in selected patients with disseminated seminoma.


Performance of entropy and Bispectral Index as measures of anaesthesia effect in children of different ages.
Entropy and Bispectral Index (BIS) have been promoted as EEG-based anaesthesia depth monitors. The EEG changes with brain maturation, but there are limited published data describing the characteristics of entropy in children, and some data suggest that BIS is less reliable in young children. The aim of this study was to compare the performance of entropy as a measure of anaesthetic effect in different age groups. The performance of entropy was compared with BIS. Fifty-four children receiving a standard sevoflurane anaesthetic for cardiac catheter studies were enrolled. The entropy and BIS were recorded pre-awakening and at 1.5%, 2% and 2.5% steady-state end-tidal sevoflurane concentrations. For analysis children were divided into four age groups: 0-1 yr, 1-2 yr, 2-4 yr and 4-12 yr. The pre-awakening values were obtained in 46 children. The median pre-awakening values for entropy and BIS varied significantly across ages with the values being lowest in the 0-1 yr age group (response entropy: 45 vs 84, 87 and 89, P=0.003; state entropy: 36 vs 78, 74 and 77, P=0.009; BIS: 56 vs 78, 76.5 and 72, P=0.02). Values were recorded at all three sevoflurane concentrations in 48 children. Compared with older groups, the 0-1 yr age group had the least significant difference in BIS and entropy when compared among different sevoflurane concentrations. The calculated sevoflurane concentrations to achieve mid-scale values of entropy and BIS were highest in the 1-2 yr age group, lower in the 0-1 yr age group and progressively lower in the 2-4 and 4-12 yr age groups. For both entropy and BIS the measure of anaesthetic effect was significantly different for children aged <1 yr compared with older children. There was no difference in performance of entropy and BIS. Both should be used cautiously in small children.


The long-term results of pre-operative hyperthermo-chemo-radiotherapy for oesophageal carcinoma--a comparison with preoperative radiation therapy alone.
The long-term results of patients with oesophageal carcinoma treated with pre-operative hyperthermo-chemo-radiotherapy (HCR) and pre-operative radiation therapy alone were compared. Twenty-six patients treated with pre-operative hyperthermo-chemo-radiotherapy (HCR Group) and 25 treated with radiation therapy alone (R Group), which demonstrated histopathologically marked effective results (Grade 3), were entered into the study. The 3-year survival rates after oesophagectomy in the HCR Group and the R Group were 67.4% and 41.8%, respectively, while the 5-year survival rates were 50.5% and 34.9%, respectively. Thus post-operative prognosis in the HCR Group was significantly more favourable than that in the R Group (P < 0.05). The local recurrence and distant metastasis rate in the HCR Group was significantly less than in the R Group (P < 0.01). This significant difference in prognosis was thought to be due to the reinforced effects of local regulation by hyperthermia and the systemic control of micrometastasis by chemotherapy in addition to radiation. Our data suggest that for carcinoma of oesophagus, pre-operative hyperthermo-chemo-radiotherapy contributes to prolonged post-operative survival while reducing both local recurrence and micrometastasis.


hMena: altered expression in psoriatic skin.
Psoriasis is a chronic inflammatory skin disease, characterized by an enhanced proliferation and a deregulated differentiation of keratinocytes. hMena is an actin regulatory protein involved in the control of cell motility and adhesion. hMena results up-modulated in several human tumors with respect to normal tissues and its expression has been positively correlated to proliferation rate, tumor size and aggressiveness in response to mitogenic stimuli, such as epidermal growth factor. The hyperproliferation of keratinocytes observed in psoriasis prompted us to evaluate hMena expression on biopsies collected from involved and uninvolved skin of 12 patients with active plaque-type psoriasis with respect to healthy skin. We analyzed the expression of hMena at transcript and protein levels by quantitative RT-PCR and immunohistochemistry. We correlated the expression of hMena to Ki67 proliferation index and to keratin 10 (K10) and keratin 16 (K16) used as markers of keratinocyte differentiation and activation. We demonstrated the expression of hMena in a hyperproliferative skin condition not related to neoplastic transformation. Interestingly, we observed that hMena is not expressed in healthy skin, but it becomes detectable in non-lesional areas and it is even more expressed in lesional psoriatic skin. In addition, we found that hMena expression is correlated to the rate of keratinocyte proliferation and activation. Hence, our observations indicate hMena as a new possible player, involved in the development and/or maintenance of the hyperproliferative state of psoriatic keratinocytes.


Cost of achieving equivalent outcomes in sicker patients after liver transplant.
We aimed to characterize variability in cost after straightforward orthotopic liver transplant (OLT). Using the University HealthSystem Consortium and Scientific Registry of Transplant Recipients databases, we identified patients who underwent OLT between 2011 and 2014. Patients meeting criteria for straightforward OLT, defined as length of stay < 14 days with discharge to home, were selected (n = 5763) and grouped into tertiles (low, medium, high) according to cost of perioperative stay. Patients undergoing straightforward OLT were of similar demographics regardless of cost. High cost patients were more likely to require preoperative hemodialysis, had higher severity of illness, and higher model for end-stage liver disease (MELD) (p < 0.01). High cost patients required greater utilization of resources including lab tests, blood transfusions, and opioids (p < 0.01). Despite having higher burden of disease and requiring increased resource utilization, high cost OLT patients with a straightforward perioperative course were shown to have identical 2-year graft and overall survival compared to lower cost patients (p = 0.82 and p = 0.63), respectively. Providing adequate perioperative care for OLT patients with higher severity of illness and disease burden requires increased cost and resource utilization; however, doing so provides these patients with long term survival equivalent to more routine patients.


The association between television viewing and irregular sleep schedules among children less than 3 years of age.
Regular sleep schedules are an important part of healthy sleep habits. Although television viewing is associated with altered sleep patterns and sleep disorders among children and adolescents, the effect of television viewing on the sleep patterns of infants and toddlers is not known. To test the hypothesis that television viewing by infants and toddlers is associated with having irregular naptime and bedtime schedules. We used data from the National Survey of Early Childhood Health, a nationally representative, cross-sectional study of the health and health care of children 4 to 35 months of age. Our main outcome measures were whether children had irregular naptime and bedtime schedules. Our main predictor was hours of television watched on a daily basis. We performed multivariate logistic regression analyses, adjusting for a variety of factors including demographic information, measures of maternal mental health, and measures of family interactions, to test the independent association of television viewing and irregular naptime and bedtime schedules. Data were available for 2068 children. Thirty-four percent of all children had irregular naptime schedules, and 27% had irregular bedtime schedules. Mean hours of television viewing per day were as follows: 0.9 hours/day (95% confidence interval [CI]: 0.8-1.0 hours/day) for children <12 months of age, 1.6 hours/day (95% CI: 1.4-1.8 hours/day) for children 12 to 23 months of age, and 2.3 hours/day (95% CI: 2.1-2.5 hours/day) for children 24 to 35 months of age. In our logistic regression model, the number of hours of television watched per day was associated with both an irregular naptime schedule (odds ratio: 1.09; 95% CI: 1.01-1.18) and an irregular bedtime schedule (odds ratio: 1.13; 95% CI: 1.04-1.24). Television viewing among infants and toddlers is associated with irregular sleep schedules. More research is warranted to determine whether this association is causal.


Isolation and characterization of a Drosophila neuropeptide gene.
We have purified a 9 amino acid amidated neuropeptide, DPKQDFMRFamide, from whole adult D. melanogaster. This peptide exhibits sequence homology to the molluscan bioactive tetrapeptide FMRFamide and is a novel member of the FMRFamide peptide family. The gene encoding DPKQDFMRFamide has been cloned and characterized. It is present in a single copy per haploid genome, is expressed as a unique 1.7 kb mRNA species, and cytologically maps to 46C on the right arm of chromosome 2. Characterization of a cDNA clone indicates that the precursor protein is 347 amino acids in length and contains 5 copies of DPKQDFMRFamide, as well as 10 additional amidated peptides exhibiting varying degrees of structural relatedness. The Drosophila DPKQDFMRFamide gene and the Aplysia FMRFamide gene are ancestrally related; however, peptides display a higher degree of homology within a species than between species, suggesting intragenic concerted evolution of these neuropeptides.


Pattern-triggered immunity restricts host colonization by endophytic fusaria, but does not affect endophyte-mediated resistance.
Fusarium oxysporum (Fo) is best known as a host-specific vascular pathogen causing major crop losses. Most Fo strains, however, are root endophytes potentially conferring endophyte-mediated resistance (EMR). EMR is a mechanistically poorly understood root-specific induced resistance response induced by endophytic or nonhost pathogenic Fo strains. Like other types of induced immunity, such as systemic acquired resistance or induced systemic resistance, EMR has been proposed to rely on the activation of the pattern-triggered immunity (PTI) system of the plant. PTI is activated upon recognition of conserved microbe-associated molecular patterns (MAMPs) of invading microbes. Here, we investigated the role of PTI in controlling host colonization by Fo endophytes and their ability to induce EMR to the tomato pathogen Fo f. sp. lycopersici (Fol). Transgenic tomato and Arabidopsis plants expressing the Fo effector gene Avr2 are hypersusceptible to bacterial and fungal infection. Here we show that these plants are PTI-compromised and are nonresponsive to bacterial- (flg22) and fungal- (chitosan) MAMPs. We challenged the PTI-compromised tomato mutants with the EMR-conferring Fo endophyte Fo47, the nonhost pathogen Fom (a melon pathogen), and with Fol. Compared to wild-type plants, Avr2-tomato plants became hypercolonized by Fo47 and Fom. Surprisingly, however, EMR towards Fol, induced by either Fo47 or Fom, was unaffected in these plants. These data show that EMR-based disease resistance is independent from the conventional defence pathways triggered by PTI, but that PTI is involved in restricting host colonization by nonpathogenic Fo isolates.


Pregnancy outcome in women with reflux nephropathy--a review of experience at the Royal Women's Hospital Brisbane, 1977-1986.
The outcome of pregnancy was examined, retrospectively, in women with a diagnosis of reflux nephropathy attending The Royal Women's Hospital from January, 1977 to August, 1986. Of 15 women, 10 were known to have had the disease antepartum and 5 were diagnosed postpartum. On the basis of plasma creatine concentration prior to pregnancy, 10 patients had 22 pregnancies with normal renal function and 5 patients had 7 pregnancies with plasma creatinine concentrations ranging from 0.12 to 0.17 mmol/l. The pregnancies of the women with normal renal function were complicated by urinary tract infections, while those with impaired renal function were associated with preeclampsia. The perinatal mortality for both groups were zero. A relatively low incidence of vesicoureteric reflux was found in 12 of the offspring who were subjected to micturating cystourethrography (mild reflux in 1 of 24 ureters).


The new era of immune checkpoint inhibition and target therapy in early-stage non-small cell lung cancer. A review of the literature.
Surgery is the best option for patients with early stage non-small cell lung cancer (NSCLC). However, the rate of local and metastatic recurrences following surgery alone is high, especially in NSCLC patients with N2 lymph node involvement. A recent American study showed that 60% of lung cancers are diagnosed in an advanced stage, and less than 20% are diagnosed in an early, resectable stage. The same study reported the 5 year survival of patients with stage IV NSCLC was 6% compared to 50% in patients with resectable NSCLC depending by stage. The addition of adjuvant or neoadjuvant chemotherapy only improves 5 year survival by 5%-10%. Recently, immunotherapy with or without chemotherapy and novel targeted therapies have yielded excellent results, in terms of both progression-free survival and overall survival, in advanced NSCLC. Published studies have shown a benefit in using immunotherapy and targeted therapy in both the adjuvant and neoadjuvant settings with many further studies still ongoing. Here we review the published data on immunotherapy and targeted therapy in the adjuvant and neoadjuvant settings in patients with operable NSCLC.