Diastolic mitral and tricuspid regurgitation by Doppler echocardiography in patients with atrioventricular block: new insight into the mechanism of atrioventricular valve closure.
The purpose of this study was to prospectively determine the incidence of diastolic mitral and tricuspid regurgitation in atrioventricular (AV) block using Doppler echocardiography. The temporal relation between mitral and tricuspid diastolic insufficiency and the diastolic murmur recorded in patients with complete heart block was also investigated. Twenty-two consecutive patients with AV block (referred to the Echo-Doppler laboratory for routine clinical studies), aged 18 to 87 years, were enrolled in the study. Eleven patients had third degree AV block and a ventricular-inhibited (VVI) pacemaker, two patients had second degree AV block, seven patients had first degree AV block, one patient had blocked premature atrial complexes and one patient had atrial flutter with 4:1 AV block. Diastolic mitral regurgitation was detected in 20 patients, and diastolic tricuspid regurgitation in 21. A mid-diastolic murmur was detected in all patients except in the three youngest. The murmur occurred before diastolic regurgitation and coincided with peak forward flow through the AV valve after atrial contraction. M-mode mitral valve echocardiograms obtained in nine patients demonstrated near closure of some portions of the mitral valve after atrial contraction. Effective closure of the valve, however, did not occur unless ventricular systole supervened. In conclusion, diastolic mitral and tricuspid regurgitation are almost universally present in patients with AV block and are associated with a diastolic murmur. The murmur coincides with forward AV valve flow. Diastolic regurgitation is silent. Effective AV valve closure is not established until ventricular systole occurs, as demonstrated by M-mode echocardiographic recording of the mitral valve.


Orphan G-protein-coupled receptors: the next generation of drug targets?
The pharmaceutical industry has readily embraced genomics to provide it with new targets for drug discovery. Large scale DNA sequencing has allowed the identification of a plethora of DNA sequences distantly related to known G protein-coupled receptors (GPCRs), a superfamily of receptors that have a proven history of being excellent therapeutic targets. In most cases the extent of sequence homology is insufficient to assign these 'orphan' receptors to a particular receptor subfamily. Consequently, reverse molecular pharmacological and functional genomic strategies are being employed to identify the activating ligands of the cloned receptors. Briefly, the reverse molecular pharmacological methodology includes cloning and expression of orphan GPCRs in mammalian cells and screening these cells for a functional response to cognate or surrogate agonists present in biological extract preparations, peptide libraries, and complex compound collections. The functional genomics approach involves the use of 'humanized yeast cells, where the yeast GPCR transduction system is engineered to permit functional expression and coupling of human GPCRs to the endogenous signalling machinery. Both systems provide an excellent platform for identifying novel receptor ligands. Once activating ligands are identified they can be used as pharmacological tools to explore receptor function and relationship to disease.


Hierarchical Sb2S3/ZnIn2S4 core-shell heterostructure for highly efficient photocatalytic hydrogen production and pollutant degradation.
In this work, a novel hierarchical 1D/2D core/shell Sb2S3-ZnIn2S4 (SB-ZIS) heterostructure with highly efficient photocatalytic activities for both hydrogen production from water and organic pollutant degradation was designed and fabricated via a simple one-step hydrothermal method. The as-prepared SB-ZIS heterostructure, where ZnIn2S4 nanosheets uniformly grew onto Sb2S3 nanorod to form a tight and large intimate contacted interface, was conducive to improve the absorption capacity of light, increase the surface area, shorten the distance of electronic transmission channels and accelerate the separation and migration of photogenerated carriers. As a result, the presented SB-ZIS composites demonstrated significantly enhanced photocatalytic performances for H2 generation and Tetracycline Hydrochloride (TCH) photodegradation. The photocatalytic H2 production rate of optimal SB-ZIS-2 sample (1685.14 μmol·g-1·h-1) was about 12.24 times as large as that of pure ZnIn2S4 (137.63 μmol·g-1·h-1). The apparent quantum efficiency (AQE) at 420 nm was up to 3.8%. In addition, the highest rate constant for TCH removal (0.514 h-1) was 20.3 and 2.89 times larger than those of pure Sb2S3 and Znln2S4, respectively. The possible reaction routes of TCH and the photocatalytic reaction mechanism of SB-ZIS sample were also discussed in detail. This work will provide some useful information for the development of dual-functional Sb2S3-based type I core-shell heterostructure with an efficient photocatalytic activity for solving environmental pollution and producing clean hydrogen energy.


A protocol for detection of mitochondrial DNA deletions: characterization of a novel deletion.
To develop a protocol capable of identifying deletions in mitochondrial DNA and use it to identify the breakpoints of a mtDNA deletion in a patient with chronic progressive external ophthalmoplegia (CPEO). Deletions in mtDNA were identified by a combination of long range PCR and Southern blotting. The precise breakpoints were determined by automated DNA sequencing. A series of DNA samples from patients with suspected mitochondrial disease was subjected to a protocol, which combines long range PCR and Southern blotting. We found a unique deletion in a patient with CPEO and we identified the precise location of this deletion through DNA sequencing. Long range PCR has the advantages of speed, minimal samples requirements, and sensitivity. Southern blotting is better able to evaluate heteroplasmy and detect duplications. We suggest a protocol that enables us to identify precisely the breakpoints in a unique mutation of mtDNA in a patient with CPEO.


In vivo glycosylation of dermal and tendon type I collagen.
Recent studies show that native collagen fibers in the extracellular space can be subject to nonenzymatic glycosylation and that the extent of such glycosylation increases in clinical hyperglycemia and aging. In the present study, a comparison was made on the extent of glycosylation in rat tail tendon and in the soluble and insoluble fractions of collagen separated from rat skin after in vivo labeling with [14C]glucose. It was observed that nonenzymatic glycosylation occurred maximally in the salt-soluble fraction as measured by the level of ketoamine linked hexose. 14C radioactivity incorporation as well as the number of free amino groups was also increased in this fraction. However, the amounts of O-glycosidically linked sugars did not show much variation between the soluble and insoluble fractions. These findings could be correlated to the enhanced metabolic turnover of newly synthesized collagen in diabetics.


An isolated, antegrade, perfused, peroneal nerve anterior tibialis muscle model in the rat: a novel model developed to study the factors governing the time course of action of neuromuscular blocking agents.
A model of an antegrade, perfused, isolated rat peroneal nerve anterior tibial muscle was developed to study potentially important factors governing the time course of action of (nondepolarizing) neuromuscular blocking agents such as concentration, blood flow, and temperature. The model allows observation of the effects of selective changes in these factors. The authors isolated the anterior tibial muscle and cannulated the anterior tibial artery and vein, providing a way for single-pass perfusion with blood from a donor rat. A force transducer was connected to the tibialis anterior muscle and a stimulator was connected to the tibial nerve. The influence of intrinsic potency (EC90) and muscle blood flow rate on the time course of pancuronium and rocuronium was investigated. The model remained stable for at least 4 h with respect to twitch height, muscle structure and function, and blood chemistry. Doubling the muscle-blood flow resulted in a significantly faster onset and offset for both pancuronium and rocuronium. Trebling the intrinsic potency (EC90) was not associated with significant changes in the time course of action of the relaxants. The authors developed and validated a model that allows us to study biophase kinetics of neuromuscular blocking agents in the anterior tibial muscle of the rat. In this model, muscle-blood flow rather than EC90 appears to predominantly determine the onset and offset time of nondepolarizing muscle relaxants.


Longitudinal distribution of macronutrients in the sediments of Jegricka watercourse in Vojvodina, Serbia.
Sediment matrices, as integral organo-mineral parts of aquatic bodies, can effectively bind and accumulate nutrients and potentially hazardous substances from diffuse and/or point sources of contamination. In this study, we analysed the longitudinal distribution of macronutrients (total N and available P and K) and the mechanical composition of the sediments of Jegricka watercourse (a part of the multi-functional Danube-Tisa-Danube canal network) known for its exposure to anthropogenic loads. The results showed that the nutrient pollution index was mostly above 1.0 (in 76%, 86% and 93% of the analysed samples for K, N and P, respectively), and the mean values for N, P and K were 2.69, 1.92 and 1.24, respectively. The average content of all nutrients and the sand fraction were significantly higher, whereas the clay fraction was considerably lower, in the sediment samples than in the adjacent arable Chernozem soil used as a benchmark. The differences in the nutrient contents and mechanical properties in the sediments were measured longitudinally (at upstream vs. downstream stations) and assessed using correlation, cluster analysis, and principal component analysis. The results suggest that the nutrient sources in the sediments as well as their transport and loading mechanisms along Jegricka watercourse are diverse and complex, likely driven by a combination of untreated industrial/urban wastewater discharges, erosion and surface runoff from the surrounding agricultural land. As a majority of the analysed watercourse banks belong to areas of special ecological value, the obtained results may be useful: i) indicators for designing and implementation of sustainable land/water policies and measures for the protection and rehabilitation of these valuable ecosystems, ii) inputs for testing/calibrating the sediment transport models and iii) the basis for sediment management.


PolyNaSS grafting on titanium surfaces enhances osteoblast differentiation and inhibits Staphylococcus aureus adhesion.
Titanium surface modifications to simultaneously prevent bacterial adhesion but promote bone-cell functions could be highly beneficial for improving implant osseointegration. In the present in vitro study, the effect of sulfonate groups on titanium surfaces was investigated with respect to both S. aureus adhesion and osteoblast functions pertinent to new bone formation. Commercial pure titanium (cpTi) squares were oxydized (Tiox), grafted with poly(sodium styrene sulfonate) groups (Tigraft) by covalent bonding using radical polymerization, and were characterized by infrared spectroscopy (HATR-FTIR) and colorimetry. Bacterial adhesion study showed that Tigraft exhibited high inhibition of S. aureus adhesion S at levels >90 %, when compared to cpTi (P < 0.05). In contrast osteoblasts adhesion was similar on all three titanium surfaces. While the kinetics of cell proliferation were similar on the three titanium surfaces, Alkaline phosphatase-specific activity of osteoblasts cultured on Tigraft surfaces was twofold higher than that observed on either on Tiox or cpTi surfaces (P < 0.01). More importantly, the amount and the distribution of calcium-containing nodules was different. The total area covered by calcium-containing nodules was 2.2-fold higher on the Tigraft as compared to either Tiox or cpTi surfaces (P < 0.01). These results provide evidence that poly(sodium styrene sulfonate) groups grafting on cpTi simultaneously inhibits bacteria adhesion but promote osteoblast function pertinent to new bone formation. Such modified titanium surfaces offer a promising strategy for preventing biofilm-related infections and enhancing osteointegration of implants in orthopaedic and dental applications.


Healthcare professionals' knowledge, attitudes, and practices regarding graduated compression stockings: a survey of China's big-data network.
The accurate identification of venous thromboembolism prophylaxis implementation barriers is an important part of prophylaxis prevention. However, in China, data to help identify these barriers is limited. This study has two objectives: 1) to determine the knowledge, attitudes, and practices (KAPs) of healthcare professionals regarding graduated compression stockings (GCS) since the launch of the National Program for the Prevention and Management of Pulmonary Embolism (PE) and Deep Venous Thrombosis (DVT) in October 2018 and 2) to identify the obstacles and assist the program. This was a cross-sectional study of 5070 healthcare professionals in China. We used exploratory factor and reliability analyses to evaluate the researcher-designed questionnaire's reliability and validity. The formal questionnaire, which included demographic data, knowledge, attitudes, and clinical practice patterns, was distributed to healthcare professionals. Of the 5070 respondents, 32.5% had a good knowledge of GCS, 78.5% had a positive attitude towards their use, and 34.0% exhibited normative behavior when applying them. The KAPs of healthcare professionals towards GCS were significantly correlated with one another. Binary logistic regression suggested that the training received by healthcare professionals was an important factor affecting their knowledge regarding GCS usage. The training provided for the use of GCS in China cannot meet medical staff needs and deserves more attention from policy makers. This represents an obstacle for venous thromboembolism prophylaxis, which restricts the effective implementation of the National Program for Prevention and Management of PE and DVT.


A new era of isotope ecology: Nitrogen isotope ratio of amino acids as an approach for unraveling modern and ancient food web.
Food web research is rapidly expanding through study of natural fractional abundance of 15N in individual amino acids. This paper overviews the principles of this isotope approach, and from my perspective, reanalyzes applications, and further extends the discussion. It applies kinetic isotope effects that enriches 15N in certain amino acids associated with the metabolic processes, which was clearly demonstrated by observations of both natural ecosystem and laboratory experiments. In trophic processes 'trophic amino acids' such as glutamic acid that significantly enrich 15N, whereas 'source amino acids' such as phenylalanine and methionine show little 15N enrichment. Through various applications conducted over the years, the principles of the method have shown to operate well and disentangle complex food webs and relevant problems. Applications include food chain length estimate, nitrogen resource assessment, tracking fish migration, and reconstruction of paleodiet. With this approach, considerations of a wide range of classical issues have been reinvigorated, while in the same time, new challenging frontiers are emerging.


Efficacy and safety of oral semaglutide by subgroups of patient characteristics in the PIONEER phase 3 programme.
To evaluate the efficacy and safety of oral semaglutide versus comparators by patient characteristic subgroups in patients with type 2 diabetes. Change from baseline in glycated haemoglobin (HbA1c) and body weight, and achievement of HbA1c <7.0% with oral semaglutide 7 mg, oral semaglutide 14 mg, flexibly dosed oral semaglutide (flex) and comparators were assessed across baseline subgroups (age, race, ethnicity, diabetes duration, body mass index and HbA1c) from the PIONEER programme. Treatment differences were analysed using a mixed model for repeated measurements for continuous variables and a logistic regression model for the binary endpoint. Pooled safety data were analysed descriptively. Changes from baseline in HbA1c and body weight, and the odds of achieving HbA1c <7.0%, were greater with oral semaglutide 14 mg/flex (n = 1934) and higher or similar with oral semaglutide 7 mg (n = 823) versus comparators (n = 2077) across most subgroups. Changes in HbA1c with oral semaglutide 14 mg/flex were greater for patients with higher baseline HbA1c (HbA1c >9.0%: -1.7% to -2.6%; HbA1c <8.0%: -0.7% to -1.2%). In some trials, Asian patients experienced greater HbA1c reductions with oral semaglutide 14 mg/flex (-1.5% to -1.8%) than other racial groups (-0.6% to -1.6%). The overall incidence of adverse events (AEs) with oral semaglutide was similar to that with comparators and was consistent across subgroups. More gastrointestinal AEs were observed with oral semaglutide, versus comparators, across subgroups. Oral semaglutide demonstrated consistently greater HbA1c and body weight reductions across a range of patient characteristics, with greater HbA1c reductions seen at higher baseline HbA1c levels.


Effect of body mass index on angle of needle insertion during ultrasound-guided lateral sagittal infraclavicular brachial plexus block.
The aim of our study was to establish the angle of needle insertion from the anterior chest wall during ultrasound-guided infraclavicular brachial plexus block and to examine for any correlation between body mass index (BMI) and insertion angle. This is a prospective observational study. The setting is at an operating room, university-affiliated teaching hospital. The patients are 23 American Society of Anesthesiologists physical status 1-3 patients scheduled to undergo elbow, forearm, or hand surgery under regional anesthesia with or without general anesthesia. The intervention is infraclavicular brachial plexus block with or without perineural catheter insertion. The measurement is the angle of needle insertion in relation to the anterior chest wall, BMI, and needle visibility as graded by the anesthesiologist. Twenty-three patients were studied. The mean (SD) BMI was 28.5 (5.4). The median (range) of angle of needle insertion was 50 (33-60). The Pearson correlation coefficient for BMI and angle of needle insertion was 0.357. There were no reported complications. The median (range) angle of needle insertion in relation to chest for our study patients was 50° (33°-60°). The needle visibility was rated difficult, requiring hydrolocation or "heeling-in," in 39% of cases. There was a moderate correlation between BMI and angle of insertion. Despite difficulties with needle visualization, the ultrasound-guided infraclavicular brachial plexus block provided reliable analgesia.


Unraveling salt responsive metabolites and metabolic pathways using non-targeted metabolomics approach and elucidation of salt tolerance mechanisms in the xero-halophyte Haloxylon salicornicum.
Haloxylon salicornicum is a xero-halophyte growing in saline and arid regions of the world. Metabolite profiling was carried out in shoot of both control and salinity treated (400 mM NaCl) samples by GC-QTOF-MS and HPLC-DAD analysis to decipher the salinity tolerance mechanism in this xero-halophyte. The present study investigates the alteration in metabolite profile of H. salicornicum that support the salinity tolerance of the plant. The metabolomic analysis of H. salicornicum shoot identified 56 metabolites, of which 47 metabolites were significantly changed in response to salinity. These metabolites were mainly included in the category of amino acids, organic acids, amines, sugar alcohols, sugars, fatty acids, alkaloids, and phytohormones. In response to salinity, most of the amino acids were down-regulated except alanine, phenylalanine, lysine, and tyramine, which were up-regulated in H. salicornicum. In contrast to amino acids, most sugars and organic acids were up-regulated in response to salinity. Correlation and pathway enrichment analysis identified important biological pathways playing significant roles in conferring salt tolerance of H. salicornicum. These biological pathways include amino sugar and nucleotide sugar metabolism, citrate cycle (TCA cycle), starch and sucrose metabolism, phenylalanine metabolism, cysteine, methionine, glycine, serine, and threonine metabolism, etc. The data presented here suggest that the modulations of various metabolic pathways facilitate H. salicornicum to survive and grow optimally even under high salinity condition. This study offers comprehensive information on metabolic adaptations and overall salt tolerance mechanisms in H. salicornicum. The information gained through this study will provide guidance to plant breeders and molecular biologists to develop salinity tolerant crop varieties.


Survival of hurricane-generated coral fragments and a disturbance model of reef calcification/growth rates.
Hurricane Gerta, with winds reaching 150 km/h, crossed the Belize barrier reef on September 18, 1978. Breakage and scouring of corals occurred in all zones of the reef to a depth of approximately 25 m. Survivorship of storm-generated coral fragments and detached colonies is strongly size dependent, conforming to the power function Y=4.44X0.66 where Y is the percent of fragments and X is the fragment size. Forty-six percent of detached Acropora palmata branches, which are larger ([Formula: see text]=37.6 cm long) than fragments of other species ([Formula: see text]=16.7 cm long), survived. Overall, 39% of fragments and detached colonies survived. This high survivorship, which probably increased the total number of colonies present, and redistribution of corals may explain the rapid recovery of reefs from all but the severest hurricanes. Storms appear to prevent coral reefs from reaching a mature state characterized by low calcification and growth rates. Therefore, we suggest that long-term reef calcification and growth rates are highest on reefs periodically distrubed by storms of intermediate intensity.


Comparing the Rates of Retinal Nerve Fiber Layer and Ganglion Cell-Inner Plexiform Layer Loss in Healthy Eyes and in Glaucoma Eyes.
To compare the rates of circumpapillary retinal nerve fiber layer (RNFL) and macular retinal ganglion cell-inner plexiform layer (GCIPL) change over time in healthy and glaucoma eyes. Cohort study. The rates of circumpapillary RNFL and macular GCIPL loss in 28 healthy subjects and 97 glaucoma subjects from the Diagnostic Innovations in Glaucoma Study (DIGS) were compared using mixed-effects models. The median follow-up time and number of visits were 1.7 years and 6 visits and 3.2 years and 7 visits for healthy and glaucoma eyes, respectively. Significant rates of loss of both global circumpapillary RNFL and average macular GCIPL thickness were detectable in early and moderate glaucoma eyes; in severe glaucoma eyes, rates of average macular GCIPL loss were significant, but rates of global circumpapillary RNFL loss were not. In glaucoma eyes, mean rates of global circumpapillary RNFL thickness change (-0.98 μm/year [95% confidence interval (CI), -1.20 to -0.76]) and normalized global circumpapillary RNFL change (-1.7%/year [95% CI, -2.1 to -1.3]) were significantly faster than average macular GCIPL change (-0.57 μm/year [(95% CI, -0.73 to -0.41]) and normalized macular GCIPL change (-1.3%/year [95% CI, -1.7 to -0.9]). The rates of global and inferior RNFL change were weakly correlated with global and inferior macular GCIPL change (r ranges from 0.16 to 0.23, all P < .05). In this cohort, the rate of circumpapillary RNFL thickness change was faster than macular GCIPL change for glaucoma eyes. Global circumpapillary RNFL thickness loss was detectable in early and moderate glaucoma, and average macular GCIPL thickness loss was detectable in early, moderate, and severe glaucoma, suggesting that structural changes can be detected in severe glaucoma.


A hierarchy of coupling free energies underlie the thermodynamic and functional architecture of protein structures.
Protein sequences and structures evolve by satisfying varied physical and biochemical constraints. This multi-level selection is enabled not just by the patterning of amino acids on the sequence, but also via coupling between residues in the native structure. Here, we employ an energetically detailed statistical mechanical model with millions of microstates to extract such long-range structural correlations, i.e. thermodynamic coupling free energies, from a diverse family of protein structures. We find that despite the intricate and anisotropic distribution of coupling patterns, the majority of residues (>70%) are only marginally coupled contributing to functional motions and catalysis. Physical origins of 'sectors', determinants of native ensemble heterogeneity in extant, ancient and designed proteins, and the basis for allostery emerge naturally from coupling free energies. The statistical framework highlights how evolutionary selection and optimization occur at the level of global interaction network for a given protein fold impacting folding, function, and allosteric outputs.


Role of oxidative stress-mediated cell death and signaling pathways in experimental fluorosis.
Free radicals and other oxidants have enticed the interest of researchers in the fields of biology and medicine, owing to their role in several pathophysiological conditions, including fluorosis (Fluoride toxicity). Radical species affect cellular biomolecules such as nucleic acids, proteins, and lipids, resulting in oxidative stress. Reactive oxygen species-mediated oxidative stress is a common denominator in fluoride toxicity. Fluorosis is a global health concern caused by excessive fluoride consumption over time. Fluoride alters the cellular redox homeostasis, and its toxicity leads to the activation of cell death mechanisms like apoptosis, autophagy, and necroptosis. Even though a surfeit of signaling pathways is involved in fluorosis, their toxicity mechanisms are not fully understood. Thus, this review aims to understand the role of reactive species in fluoride toxicity with an outlook on the effects of fluoride in vitro and in vivo models. Also, we emphasized the signal transduction pathways and the mechanism of cell death implicated in fluoride-induced oxidative stress.


Evidence that multiple risk factors for coronary artery disease exist in persons with abnormal glucose tolerance.
Multiple risk factors for coronary artery disease were determined in 50 healthy, non-diabetic persons with an oral glucose tolerance test result that could not be classified as normal by current criteria and 50 sex-, age-, and weight-matched persons with normal oral glucose tolerance. The results indicated that persons with abnormal oral glucose tolerance were hyperinsulinemic, as well as hypercholesterolemic and hypertriglyceridemic. In addition, patients with abnormal results in glucose tolerance tests had significantly elevated systolic blood pressure and heart rates. These data suggest that a cluster of risk factors for coronary artery disease exists in non-diabetic persons with abnormal oral glucose tolerance.


Sandwich therapy in testis tumor: current experience.
Sandwich therapy, a regimen of lymphadenectomy combined with preoperative and postoperative cobalt teletherapy, yielded a survival rate of 84% in 13 patients with stages A and B non-seminomatous testis tumor. This statistic is consistent with the previously reported survival rate of 83% achieved among 35 patients treated with this regimen from 1958 to 1970 at our institution. Bone marrow depression, retroperitoneal fibrosis and possible induction of a second malignancy were important side effects of radiation therapy. The superior survival rates with fewer severe side effects of treatment obtained by other investigators using either lymphadenectomy alone or lymphadenectomy combined with chemotherapy have prompted us to discontinue sandwich therapy as standard treatment for non-seminomatous testis tumors at our institution.


Positive skin reaction induced by 4,4'-azoxybenzenedisulfonamide in relationship to the sulfanilamide allergy.
All guinea pigs sensitized to sulfanilamide or 4-hydroxylaminobenzenesulfonamide in the absence of ultraviolet irradiation exhibited positive skin reactions to 4,4'-azoxybenzenedisulfonamide, 4-nitrosobenzenesulfonamide, and 4-hydroxylaminobenzenesulfonamide.


Comparison of the pharmacokinetics of naproxen tablets and suspension in children.
Twenty-three children, aged 8 to 14 years, with postoperative pain, were randomly assigned to receive a fixed 250-mg dose (4.66-7.58 mg/kg) of naproxen as either a liquid suspension or tablet. After an overnight fast, the serum concentrations were measured before and at 0.5, 1, 2, 3, 4, 8, 12, 18, and 24 hours after administration of naproxen. The concentration versus time data were best fit to a one-compartment open model. The area under the concentration versus time curve, apparent volume of distribution (VDss/F), and elimination parameters (CL/F, Ke, elimination half-life) were similar in children who received suspension or tablets. Although the apparent maximum peak plasma concentration (Cmax) was greater in children who received tablets compared with those who received the suspension, Cmax/area under the curve (AUC), apparent time to maximum peak concentration (tmax), Ka, and estimated time to 10%, 50%, and 90% absorption (T10, T50, T90) were not different. The dose range was relatively narrow; hence, direct relationships between dose and elimination parameters, VDss/F, apparent tmax, Ka, T10, T50 or T90 were not observed. Neither VDss/F or CL/F were age related over the relatively narrow range of ages that were studied. Elimination of naproxen in our patients was more rapid than has previously been reported in children or adults, however. From a practical standpoint, naproxen tablets and suspension seem to be bioequivalent in fasting children ages 8 to 14 years.


Identification of two Drosophila TGF-beta family members in the grasshopper Schistocerca americana.
Intercellular signaling molecules of the transforming growth factor-beta (TGF-beta) superfamily are required for pattern formation in many multicellular organisms. The decapentaplegic (dpp) gene of Drosophila melanogaster has several developmental roles. To improve our understanding of the evolutionary diversification of this large family we identified dpp in the grasshopper Schistocerca americana. S. americana diverged from D. melanogaster approximately 350 million years ago, utilizes a distinct developmental program, and has a 60-fold-larger genome than D. melanogaster. Our analyses indicate a single dpp locus in D. melanogaster and S. americana, suggesting that dpp copy number does not correlate with increasing genome size. Another TGF-beta superfamily member, the D. melanogaster gene 60A, is also present in only one copy in each species. Comparison of homologous sequences from D. melanogaster, S. americana, and H. sapiens, representing roughly 900 million years of evolutionary distance, reveals significant constraint on sequence divergence for both dpp and 60A. In the signaling portion of the dpp protein, the amino acid identity between these species exceeds 74%. Our results for the TGF-beta superfamily are consistent with current hypotheses describing gene duplication and diversification as a frequent response to high levels of selective pressure on individual family members.


Basic mechanisms of atrial fibrillation.
The mechanism of AF remains controversial as support still exists for multiple wavelets, "mother rotor," and focal sources. These mechanisms need not be mutually exclusive. For example, the mother rotor hypothesis may not be distinct form the focal AF, if the rotor is of small size (ie, microre-entry). With the development of several animal models to study AF, along with improving technologies and mapping techniques, a further understanding of the pathophysiology of AF is being gained. Each animal model has unique electrophysiological and structural abnormalities, and one may not be able to generalize from one model to the next. It is likely that there is not one mechanism for all AF, but that there are substrate-specific mechanisms, and that AF may be comprised of several different mechanisms.


Discovery of a novel cyclin-dependent kinase 8 inhibitor with an oxindole core for anti-inflammatory treatment.
Chronic inflammation is an underlying cause in a number of diseases. Cyclin-dependent kinase 8 (CDK8) has been implicated as an inflammatory mediator, indicating its potential as an anti-inflammatory target. Herein, we performed structure-based virtual screening (SBVS) to identify novel CDK8 inhibitors. The pharmacological interactions for CDK8 were identified and incorporated into a SBVS protocol. Selected compounds were tested in enzymatic assays, and one compound was confirmed to be a CDK8 inhibitor with a 50% inhibitory concentration (IC50) value of 1684.4 nM. Comparing structural analogs identified a compound, F059-1017, with greater potency (IC50 558.1 nM). When tested in cell lines, the compounds displayed low cytotoxicity. Cellular assays revealed that the identified CDK8 inhibitors can reduce phosphorylation and expression of signaling mediators associated with inflammation. In addition, results of kinase profiling showed that compound F059-1017 is selective towards CDK8. These findings suggest that the new inhibitors have great potential as lead compounds for developing novel anti-inflammatory therapeutics.


Thermostability of heterophile antibodies from human sera infected with Schistosoma mansoni and geo-helminths. An immuno-metric statistical analysis.
Antibody in human sera that induces lysis of sheep erythrocytes in hemolytic assay was investigated. The present study showed that the presence in serum of the thermostable cytolytic anti-sheep red blood cells antibodies is dependent on the Schistosoma mansoni infection, and this is more frequent in adults than in children. The thermostable characteristic of hemolysins in normal sera was not dependent on the presence of Ascaris lumbricoides, Trichuris trichiura or hookworm geo-helminths. Further, thermostable complement-activating heterophile antibodies were noticed in children in association with massive number of S. mansoni eggs. The results were obtained by using the z- and the chi-square tests. The z-test allows us to formulate a one-sided alternative, i.e., a tendency of one of the attributes. On the other hand, the chi-square test analyzes the independence between attributes by using a contingency table. Besides the obtained results being interesting in the field of schistosomiasis mansoni, they can provide a new insight into the use of statistics in medical science.


Synthesis of Saccharomyces cerevisiae invertase by Schizosaccharomyces pombe.
In order to gain information on the ability of the glycosylation system of Schizosaccharomyces pombe to process heterologous glycoproteins, the expression of Saccharomyces cerevisiae invertase in the former yeast was studied. Sc. pombe cells are able to produce enzymatically active invertase from the S. cerevisiae SUC2 gene introduced by transformation and the enzyme is glycosylated and secreted into the cell wall. However, Sc. pombe transformants do not glycosylate the heterologous enzyme as their own invertase since it is not bound by the lectin from Bandeiraea simplicifolia seeds, which indicates the absence of terminal galactose residues. Moreover, the electrophoretic mobility of the heterologous invertase is similar to that of the large enzyme from S. cerevisiae, both in its native form and after being deglycosylated with Endo H. These results suggest that the polypeptide chain of S. cerevisiae invertase is the primary factor for the glycosylation in Sc. pombe cells.


Superoxide dismutase and glutathione peroxidase activities in erythrocytes as indices of oxygen loading in disease: a survey of one hundred cases.
It has been established that the pyrogallol autoxidation method for the estimation of the activity of superoxide dismutase (SOD) (EC 126.96.36.199) is superior in precision and sensitivity to a superoxide-generating method (NADH/phenazine methosulfate linked to nitroblue tetrazolium reduction). Reference intervals were established in an urban population in the Far East for SOD activity in erythrocytes using the pyrogallol method, and for glutathione peroxidase (GSH-Px) (EC 188.8.131.52) activity in erythrocytes using a standard glutathione reductase-linked method. On this basis, erythrocyte SOD activities were significantly (P less than 0.05) depressed in cases of visceral cancer, acute myocardial infarct, congestive heart failure, respiratory failure, chronic renal failure, and diabetes mellitus, but within the reference interval in cases of lung cancer and asthma. Erythrocyte GSH-Px activity was significantly (P less than 0.05) depressed in cases of diabetes mellitus and chronic renal failure but elevated in respiratory failure and asthma. GSH-Px and SOD activities were well correlated in patients but not in the reference population.


ProACTH: use for early detection of lung cancer.
ProACTH was identified by radioimmunoassay and column chromatography in extracts of 38 lung carcinomas. One hundred patients were evaluated because of abnormalities on chest roentgenograms; 53 (72 per cent) of 74 with lung cancers had increased plasma ACTH immunoreactivity; all 26 with benign abnormalities had normal ACTH. Of 101 patients with obstructive lung disease, five of 20 with elevated plasma ACTH levels and two of 81 with normal plasma ACTH levels had lung cancer within two years. In three of 31 patients with granulomatous lung disease plasma ACTH levels were elevated during acute exacerbation of the disease, but they returned to normal after recovery. We conclude that plasma ACTH immunoreactivity is frequently increased early in the course of lung carcinoma.


Second harmonic generation microscopy in zebrafish.
Modern optical imaging has progressed rapidly with the ability to noninvasively image cellular and subcellular phenomena with high spatial and temporal resolution. In particular, emerging techniques such as second harmonic generation (SHG) microscopy can allow for the monitoring of intrinsic contrast, such as that from collagen, in live and fixed samples. When coupled with multiphoton fluorescence microscopy, SHG can be used to image interactions between cells and the surrounding extracellular environment. There is recent interest in using these approaches to study inflammation and wound healing in zebrafish, an important model for studying these processes. In this chapter we present the practical aspects of using second harmonic generation to image interactions between leukocytes and collagen during wound healing in zebrafish.


Modulation of cortical activity after anodal transcranial direct current stimulation of the lower limb motor cortex: a functional MRI study.
Functional magnetic resonance imaging (fMRI) has shown that transcranial direct current stimulation (tDCS) of the hand motor cortex modulates cortical activity of the healthy human brain. However, few studies have assessed the effects of tDCS on the leg motor cortex. We therefore used fMRI to examine the modulating effects of tDCS on lower limb motor cortex responses. In this sham-controlled case-control study, 11 subjects were exposed to active anodal (n = 6) or sham (n = 5) stimulation, with the anode being positioned on the leg motor cortex of the right hemisphere. Each tDCS was delivered for 15 minutes at 2 mA, with each subject receiving a total of four stimulatory sessions on consecutive days. Cortical activity was measured before the first and after the fourth session by fMRI, and changes in cortical activity were calculated. Anodal tDCS increased activation of the ipsilateral supplementary motor area and lowered the extent of activation of both anterior cingulate gyri, the right middle and superior temporal gyri, the middle and superior frontal gyri, and the primary and secondary somatosensory cortices. Anodal tDCS increased corticospinal excitability of the lower limb motor cortex in healthy subjects, suggesting that multiple brain cortical areas may be associated with leg motor performance via involvement of variable corticocortical connections.


The assessment of undergraduate bachelor of nursing students in the collaborative clusters education model: A qualitative descriptive design.
Worldwide, undergraduate Bachelor of Nursing students are required to complete experiential learning placements in health care settings as part of the curriculum. There are a variety of facilitation models that support student learning and assessment on clinical placement. As workforce pressures increase globally, innovative approaches to clinical facilitation are required. In the Collaborative Clusters Education Model of clinical facilitation, hospital-employed clinical facilitators work collaboratively within peer groups (clusters) to collectively participate in a process of facilitating student learning and conducting assessment and moderation of student performance. The assessment process in this collaborative clinical facilitation model is not well described. To describe how the assessment of undergraduate nursing students is achieved in the Collaborative Clusters Education Model. A qualitative descriptive approach was employed. In March 2021 individual and group interviews were conducted with seven clinical facilitators working in the Collaborative Clusters Education Model in one health service in southeast Queensland, Australia. Content analysis of transcribed interviews was performed. Assessment was achieved through two processes, situational scoring and moderation. In the process of situational scoring, clinical facilitators balanced the students' perception of their role in assessment, accounted for the type of experiences available, considered multiple sources of evidence and used the Australian Nursing Standards Assessment Tool. In the process of moderation, clinical facilitators communicated with their cluster colleagues to determine a shared understanding of student history, considered data from multiple evidence sources and collaboratively evaluated the trustworthiness of student performance evaluation decisions. In the Collaborative Clusters Education Model, the input of multiple assessors, working in a small team, ensured transparency in assessment processes. Furthermore, this transparency in assessment practices normalised on-going moderation, an in-built quality-check and, as such, an innovative component of assessment in the Collaborative Clusters Education Model. As nursing directors and managers seek to ameliorate the impact of nursing workforce pressures, this innovative model of collaborative assessment may serve as a valuable addition to nursing clinical assessment toolkits. The Collaborative Clusters Education Model of Clinical Facilitation enables transparency in assessment processes and normalises moderation.


Association between hemoglobin glycation index and non-alcoholic fatty liver disease in the patients with type 2 diabetes mellitus.
The hemoglobin glycation index (HGI) represent the disparity between actual glycated hemoglobin measurements and predicted HbA1c. It serves as a proxy for the degree of non-enzymatic glycation of hemoglobin, which has been found to be positively correlated with diabetic comorbidities. In this study, we investigated the relationship between HGI and non-alcoholic fatty liver disease (NAFLD), along with other relevant biological markers in patients with diabetes. This cross-sectional study consisted of 3,191 adults diagnosed with type 2 diabetes mellitus. We calculated the predicted glycated hemoglobin levels based on fasting blood glucose levels. Multivariate binary logistic regression analysis was conducted to examine the correlation between the HGI and NAFLD. Hepatic steatosis was diagnosed using ultrasonography. Among all participants, 1,784 (55.91%) were diagnosed with NAFLD. Participants with confirmed NAFLD showed elevated body mass index, diastolic blood pressure, liver enzyme, total cholesterol, triglyceride, low-density lipoprotein and uric acid levels compared with those without NAFLD. In the unadjusted model, participants in the last tertile of HGI were 1.40-fold more likely to develop NAFLD than those in the first tertile (95% confidence interval 1.18-1.66; P < 0.001). In the fully adjusted model, those in the last tertile of HGI had a 39% increased risk of liver steatosis compared with confidence interval in the first tertile of HGI (95% confidence interval 1.12-1.74; P < 0.001). A higher HGI suggests an elevated risk of developing NAFLD in patients with type 2 diabetes.


Effects of chemotherapy agents on Sphingosine-1-Phosphate receptors expression in MCF-7 mammary cancer cells.
Sphingosine-1-phosphate (S1P) is a potent bioactive sphingolipid involved in the regulation of cell proliferation and cancer progression. Increased expression of S1P receptors has been detected in advanced breast tumours with poor prognosis suggesting that S1P receptors might control tumour response to chemotherapy. However, it remains unclear how the levels of S1P receptor expression are influenced by chemotherapy agents. Western immunoblotting, PCR analysis and fluorescent microscopy techniques were used in this study to analyze expression patterns of S1P receptors 2 and 3 (S1P2/S1P3) in MCF-7 breast adenocarcinoma cells treated by Tamoxifen (TAM) and/or Medroxyprogesterone acetate (MPA). We found that TAM/MPA induce downregulation of S1P3 receptors, but stimulate expression of S1P2. According to cell viability and caspase activity analyses, as expected, TAM activated apoptosis. We also detected TAM/MPA-induced autophagy marked by formation of macroautophagosomes and increased level of Beclin 1. Combined application of TAM and MPA resulted in synergistic apoptosis- and autophagy-stimulating effects. Assessed by fluorescent microscopy with autophagosome marker LAMP-2, changes in S1P receptor expression coincided with activation of autophagy, suggestively, directing breast cancer cells towards death. Further studies are warranted to explore the utility of manipulation of S1P2 and S1P3 receptor expression as a novel treatment approach.


[Energy metabolism of the cancer cell: example of mitochondria-rich endocrine tumors].
Most solid tumours preferentially develop glycolytic metabolism, often accompanying tumor aggressiveness. Increase in nucleic acid synthesis is associated with cell proliferation and glucose shunting to the pentose phosphate pathway. High glucose consumption is more associated with a metabolic adaptation than with a mitochondrial defect. Tumor cells do not present specific genetic modifications but adapt their metabolic capacities to their priority needs. However their metabolisms depend on oncogene expression more specifically expressed in this context. The glycolytic pathway is favored by tumor proliferation under hypoxia. Stabilization of HIF1 factor may explain the glycolytic metabolism of the tumors in an anaerobic environment. We demonstrate in two types of mitochondrial rich tumors, that specific defects induce completely different metabolic directions: when familial paragangliomas present a glycolytic metabolism, thyroid oncocytic tumors develop a specific oxidative metabolism.


Investigation of correlation between H63D and C282Y mutations in HFE gene and serum Ferritin level in beta-thalassemia major patients.
Mutations in the HFE gene have been shown to be associated with hemochromatosis which is observed in beta-thalassemia major. In this study, we determined the HFE gene mutations (C282Y and H63D) among b-thalassemia major patients to investigate the effect of these mutations on serum Ferritin levels. In this cross-sectional study, a total of 105 b-thalassemia subjects with a history of regular blood transfusion were selected. They divided into two distinct groups according cut off 1000ng/ml of serum Ferritin levels. The HFE gene mutant allele detected by RFLP-PCR. Of 105 thalassemia patients, 29 patients (14 male and 15 female) were heterozygote for H63D mutation, and just one male was homozygote, but for C282Y mutation just one heterozygote and one homozygote was detected, and overall 31% had coexistence of b-thal and HFE gene mutations. As expected, Ferritin levels significantly differed between groups (P=0.001). The impact of detection of HFE mutations could prognosis the likelihood of iron overload in multi-transfused patients, and allowing early diagnosis and proper management to overcome complications of iron overload in beta-thalassemia patients.


Migrant mothers and the role of social support when child rearing.
To raise children in a new country without the support of family and community may prove problematic for some women. This study explored the possible impact of a lack supportive social network on mothering. A cross sectional design using both quantitative (questionnaire) and qualitative approaches (interview) was used to investigate the child rearing experience of migrant women from the United Kingdom (UK) (N = 154) currently living in Western Australia (WA); 40 women were selected for interview. Bowlby's (1969) mother-infant attachment theory provides the theoretical framework to investigate the perceived loss of attachment to the homeland, attachment figure, country, culture and family. The results indicated that migrant women with children missed the close support of family networks; and indicate that all migrant mothers, regardless of origin, require a social support network to survive.


Lactate and pyruvate stimulate the conversion of glucose to glycogen in hepatocytes by a mechanism that does not involve gluconeogenic flux.
Glycogen synthesis in hepatocytes was determined at various concentrations of CO2 and medium HCO3- to modulate cell pH. Glycogen synthesis from glucose was highest in acidic conditions (5% CO2/12.5 mM HCO3-) and lowest in alkaline conditions (2.5% CO2/25 mM HCO3-). Physiological concentrations of lactate/pyruvate (2 mM/0.2 mM) stimulated the conversion of glucose to glycogen in all media examined and mercaptopicolinate, an inhibitor of gluconeogenesis, caused a similar stimulation as lactate/pyruvate. In alkaline media, the stimulation by mercaptopicolinate and by lactate/pyruvate was additive, indicating that the latter is not due to gluconeogenic flux. In acidic media, the stimulation by both lactate/pyruvate and mercaptopicolinate was inhibited by amiloride, an inhibitor of Na+/H+ exchange. Since Na+/H+ exchange is activated when cell pH falls below a certain threshold, it is postulated that lactate and pyruvate stimulate the conversion of glucose to glycogen through cellular acidification.


The normal cellular prion protein (PrPc) is strongly expressed in bovine endocrine pancreas.
Expression of the cellular prion protein (PrP(c)) has been shown to be crucial for the development of transmissible spongiform encephalopathies and for the accumulation of the disease-associated conformer (PrP(sc)) in the brain and other tissues. One of the emerging hypotheses is that the conversion phenomenon could take place at the site where the infectious agent meets PrP(c). In this work we have studied whether PrP(c), a protein found predominantly in neurons, could also exist in pancreatic endocrine cells since neuroectoderm-derived cells and pancreatic islet cells share a large number of similarities. For this purpose we have examined the expression of PrP(c) in a series of fetal and postnatal bovine pancreatic tissue by immunohistochemistry and RT-PCR. Using immunostained serial sections and specific antibodies against bovine PrP(c), insulin, glucagon, somatostatin, chromogranin A and chromogranin B we found that PrP(c) is highly expressed in all endocrine cells of fetal and adult pancreatic islets with a particular strong expression in A-cells. Moreover it became evident that the PrP(c) gene-neighbour chromogranin B as well as chromogranin A are coexpressed together with PrP(c). The selective expression of PrP(c) in the bovine endocrine pancreas is of particular importance regarding possible iatrogenic transmission routes and demonstrates also that bovine pancreatic islet cells could represent an interesting model to study the control of PrP-gene expression.


Canadian study of health and aging: study methods and prevalence of dementia.
To estimate the prevalence of dementia and its subtypes by sex and age group for five regions of Canada. Prevalence survey. Community and institutional settings in Canada, excluding those in the two territories, Indian reserves and military units. Representative sample of people aged 65 and over interviewed between February 1991 and May 1992. Those in the community (9008 subjects) were chosen randomly from medicare lists in nine provinces or from the Enumeration Composite Record in Ontario. People in institutions (1255) were randomly selected from residents in stratified random samples of institutions in each region. Screening with the Modified Mini-Mental State (3MS) Examination to identify cognitive impairment. Clinical examination of all those in institutions, those in the community with a 3MS score of less than 78 and a sample of those in the community with a 3MS score of 78 or more to diagnose dementia. Dementia and Alzheimer's disease were defined according to established criteria. Prevalence of dementia of all types, by region, sex and age group, the estimated number of cases in the population by type of dementia and the age-standardized rate per 1000 population. The prevalence estimates suggested that 252,600 (8.0%) of all Canadians aged 65 and over met the criteria for dementia (95% confidence interval [CI] 236,800 to 268,400). These were divided roughly equally between the community and institutional samples; the female:male ratio was 2:1. The age-standardized rate ranged from 2.4%, among those aged 65 to 74 years, to 34.5%, among those aged 85 and over. The corresponding figures for Alzheimer's disease were 5.1% overall (161,000 cases; 95% CI 148,100 to 173,900), ranging from 1.0% to 26.0%; for vascular dementia it was 1.5% overall, ranging from 0.6% to 4.8%. If the prevalence estimates remain constant, the number of Canadians with dementia will rise to 592,000 by 2021. These Canadian estimates of the prevalence of dementia fall toward the upper end of the ranges in other studies, whereas the estimates for Alzheimer's disease fall in the middle of the ranges. This may suggest an unusual balance between Alzheimer's and other forms of dementia in the Canadian population.


Enveloped viruses disable innate immune responses in dendritic cells by direct activation of TAM receptors.
Upon activation by the ligands Gas6 and Protein S, Tyro3/Axl/Mer (TAM) receptor tyrosine kinases promote phagocytic clearance of apoptotic cells and downregulate immune responses initiated by Toll-like receptors and type I interferons (IFNs). Many enveloped viruses display the phospholipid phosphatidylserine on their membranes, through which they bind Gas6 and Protein S and engage TAM receptors. We find that ligand-coated viruses activate TAM receptors on dendritic cells (DCs), dampen type I IFN signaling, and thereby evade host immunity and promote infection. Upon virus challenge, TAM-deficient DCs display type I IFN responses that are elevated in comparison to wild-type cells. As a consequence, TAM-deficient DCs are relatively resistant to infection by flaviviruses and pseudotyped retroviruses, but infection can be restored with neutralizing type I IFN antibodies. Correspondingly, a TAM kinase inhibitor antagonizes the infection of wild-type DCs. Thus, TAM receptors are engaged by viruses in order to attenuate type I IFN signaling and represent potential therapeutic targets.


Magnetocardiographic assessment of healed myocardial infarction.
We evaluated the capability of multichannel magnetocardiography (MCG) to detect healed myocardial infarction (MI). Multichannel MCG over frontal chest was recorded at rest in 21 patients with healed MI, detected by cine- and contrast-enhanced magnetic resonance imaging, and in 26 healthy controls. Of the 21 MI patients, 11 had non-Q wave and 10 Q wave MIs. QRS, ST-segment, T wave and ST-T wave integrals, ST-segment and T wave amplitudes, and QRS and ST-T wave magnetic field map orientations were measured. The MCG repolarization indexes, such as ST segment and ST-T wave integrals, separated the MI group from the controls (ST-T wave integral -1.4 +/- 5.3 vs 1.5 +/- 4.7 pTs, P = 0.034). The abnormalities were more distinct in the Q wave-MI than in the non-Q wave MI subgroup. In the latter, however, a trend similar to the Q wave MI group was found. The relation of QRS area to ST segment and T wave integral improved the detection of healed MIs compared to the ST-T wave indexes alone (QRS-ST-T discordance 14 +/- 10 vs 5.0 +/- 7.1 pTs, P = 0.003). When comparing the MI group to the controls, the orientation of the magnetic field maps differed in the ST-T wave maps (163 +/- 119 degrees vs 58 +/- 17 degrees, P < 0.001) but not in the QRS maps (111 +/- 95 degrees vs 106 +/-93 degrees, P = 0.646). The MCG repolarization variables can detect healed MI. These ST-T wave abnormalities are more pronounced in patients with Q wave MI than in patients with non-Q wave MIs. Relating the signals of depolarization and repolarization phases improves the detection of healed MI. Repolarization abnormalities are common in healed MI and thus should not always be interpreted as present ongoing ischemia.


Insights into cell-free therapeutic approach: Role of stem cell "soup-ernatant".
Current advances in medicine have revolutionized the field of regenerative medicine dramatically with newly evolved therapies for repair or replacement of degenerating or injured tissues. Stem cells (SCs) can be harvested from different sources for clinical therapeutics, which include fetal tissues, umbilical cord blood, embryos, and adult tissues. SCs can be isolated and differentiated into desired lineages for tissue regeneration and cell replacement therapy. However, several loopholes need to be addressed properly before this can be extended for large-scale therapeutic application. These include a careful approach for patient safety during SC treatments and tolerance of recipients. SC treatments are associated with a number of risk factors and require successful integration and survival of transplanted cells in the desired microenvironment with concurrent tissue regeneration. Recent studies have focused on developing alternatives that can replace the cell-based therapy using paracrine factors. The development of stem "cell free" therapies can be devoted mainly to the use of soluble factors (secretome), extracellular vesicles, and mitochondrial transfer. The present review emphasizes on the paradigms related to the use of SC-based therapeutics and the potential applications of a cell-free approach as an alternative to cell-based therapy in the area of regenerative medicine.


Arsenic and fluoride in the groundwater of Mexico.
Concentrations of arsenic and fluoride above Mexican drinking water standards have been detected in aquifers of various areas of Mexico. This contamination has been found to be mainly caused by natural sources. However, the specific processes releasing these toxic elements into groundwater have been determined in a few zones only. Many studies, focused on arsenic-related health effects, have been performed at Comarca Lagunera in northern México. High concentrations of fluoride in water were also found in this area. The origin of the arsenic there is still controversial. Groundwater in active mining areas has been polluted by both natural and anthropogenic sources. Arsenic-rich minerals contaminate the fractured limestone aquifer at Zimapán, Central México. Tailings and deposits smelter-rich fumes polluted the shallow granular aquifer. Arsenic contamination has also been reported in the San Antonio-El Triunfo mining zone, southern Baja California, and Santa María de la Paz, in San Luis Potosí state. Even in the absence of mining activities, hydrogeochemistry and statistical techniques showed that arsenopyrite oxidation may also contaminate water, as in the case of the Independencia aquifer in the Mexican Altiplano. High concentrations of arsenic have also been detected in geothermal areas like Los Azufres, Los Humeros, and Acoculco. Prevalence of dental fluorosis was revealed by epidemiological studies in Aguascalientes and San Luis Potosí states. Presence of fluoride in water results from dissolution of acid-volcanic rocks. In Mexico, groundwater supplies most drinking water. Current knowledge and the geology of Mexico indicate the need to include arsenic and fluoride determinations in groundwater on a routine basis, and to develop interdisciplinary studies to assess the contaminant's sources in all enriched areas.


Antioxidants in fertility: impact on male and female reproductive outcomes.
A couple may be considered to have fertility problems if they have been trying to conceive for over 1 year with no success. Worldwide, the inability to have children affects 10% to 15% of all couples. Subfertility can be divided into either male or female factor, or both partners can be affected. However, for some couples the cause for subfertility cannot be identified, and this is called unexplained subfertility. It is thought that oxidative stress is involved in the pathophysiology of subfertility, and antioxidants are thought to reduce the damage caused by oxidative stress. Antioxidants are widely available and inexpensive. However, there is currently little high-quality evidence to show that taking antioxidants will provide any benefit or harm for infertile couples.


Linking key human-environment theories to inform the sustainability of coral reefs.
Effective solutions to the ongoing "coral reef crisis" will remain limited until the underlying drivers of coral reef degradation are better understood. Here, we conduct a global-scale study of how four key metrics of ecosystem states and processes on coral reefs (top predator presence, reef fish biomass, trait diversity, and parrotfish scraping potential) are explained by 11 indicators based on key human-environment theories from the social sciences. Our global analysis of >1,500 reefs reveals three key findings. First, the proximity of the nearest market has the strongest and most consistent relationships with these ecosystem metrics. This finding is in keeping with a body of terrestrial research on how market accessibility shapes agricultural practices, but the integration of these concepts in marine systems is nascent. Second, our global study shows that resource conditions tend to display a n-shaped relationship with socioeconomic development. Specifically, the probabilities of encountering a top predator, fish biomass, and fish trait diversity were highest where human development was moderate but lower where development was either high or low. This finding contrasts with previous regional-scale research demonstrating an environmental Kuznets curve hypothesis (which predicts a U-shaped relationship between socioeconomic development and resource conditions). Third, together, our ecosystem metrics are best explained by the integration of different human-environment theories. Our best model includes the interactions between indicators from different theoretical perspectives, revealing how marine reserves can have different outcomes depending on how far they are from markets and human settlements, as well as the size of the surrounding human population.


Volume of RBCs, 24- and 48-hour posttransfusion survivals, and the lifespan of (51)Cr and biotin-X-N-hydroxysuccinimide (NHS)-labeled autologous baboon RBCs: effect of the anticoagulant and blood pH on (51)Cr and biotin-X-NHS elution in vivo.
The RBC injury that occurs during collection of the first few milliliters of blood into the pH 5.0 ACD (NIH, Formula A) is referred to as the lesion of collection. The RBC injury was evaluated by labeling the ACD RBCs with (51)Cr and measuring the 24-hour posttransfusion survival. The effect of the acidification of ACD blood on the in vivo elution of (51)Cr from the RBC has not been reported. Baboon blood was collected in heparin and in ACD and CP2D at different ratios of blood to anticoagulant. ACD blood with a pH of 5.7 to 6.9 was labeled with (51)Cr. Heparinized blood with a pH of 7.4 was labeled with biotin-X-N-hydroxysuccinimide (NHS). ACD blood with a pH of 5.9 was labeled with both (51)Cr and biotin-X-NHS. The RBC volumes, 24- and 48-hour posttransfusion survivals, and lifespans (T50) were measured. The RBC volume of ACD blood with a pH ranging from 5.7 to 6.9 was not affected by (51)Cr labeling. (51)Cr-labeled ACD blood with a pH of 6.9 had an RBC volume that was significantly greater than that seen in heparinized blood with a pH of 7.4 labeled with biotin-X-NHS. In vivo elution of (51)Cr from the RBCs prepared from the ACD blood with a pH of 5.7 to 6.9 and in vivo elution of biotin-X-NHS from RBCs prepared from ACD blood with a pH of 5.9 were associated with reductions in 24- and 48-hour posttransfusion survivals and T50. The anticoagulant and the pH of the medium in which the RBCs were labeled with (51)Cr or biotin-X-NHS affected in vivo elution of the label from the RBCs and may reduce posttransfusion survival values.


The influence of front-of-pack portion size images on children's serving and intake of cereal.
Consumption of large portions of energy-dense foods promotes weight gain in children. Breakfast cereal boxes often show portions much larger than the recommended serving size. This experimental study investigated whether front-of-package portion size depictions influence children's self-served portions and consumption. In a between-subjects design, 41 children aged 7-11 years (M= 9.0 ± 1.5y) served themselves breakfast cereal from a box, the front of which depicted either a recommended serving size of cereal (30g) or a larger, more typical front-of-pack portion (90g). Cereal served and consumed and total caloric intake (including milk) was recorded. Height and weight, demographic information and measures of children's food responsiveness and enjoyment of food were collected. MANOVA revealed that children exposed to the larger portion size served themselves (+7g, 37%) and consumed (+6g, 63%) significantly more cereal than those exposed to the smaller portion. Despite this, overall caloric intake (milk included) did not differ between conditions, and no other measured variables (hunger, BMI) significantly affected the outcomes. This study provides novel evidence of the influence portion-size depictions on food packaging have on children's eating behaviour. This offers possible avenues for intervention and policy change; however, more research is needed.


Reevaluation of the evidence that an antibody to the insulin receptor is insulinmimetic without activating the protein tyrosine kinase activity of the receptor.
The immunoglobulin fraction of a polyclonal anti-insulin receptor antibody (B-10) derived from a patient with severe insulin resistance and acanthosis nigricans was tested for its ability to activate the protein kinase activity of the insulin receptor and to mimic insulin action in Chinese hamster ovary cells expressing either wild type or kinase-deficient human insulin receptors. This antiserum had previously been reported to be insulinmimetic without activating the insulin receptor protein tyrosine kinase. Antibody B-10 bound to both wild type and mutant human insulin receptors, but it induced receptor down-regulation and stimulated hexose transport and thymidine incorporation into DNA only in cells expressing the wild type receptor. Furthermore, this antibody activated the kinase activity of the wild type insulin receptor in intact cells and in vitro. It is likely, therefore, that the biological activities of antibody B-10, like those of insulin, depend upon the protein tyrosine kinase activity of the insulin receptor.


Dissolution studies on TiO2 with organics.
In this work the effect of organic reducing reagents, namely, ascorbic acid, oxalic acid and L-cysteine on dissolution of commercial TiO(2) has been investigated. Kinetic studies showed that a maximum of about 45% of TiO(2) was dissolved by ascorbic acid in 4h when oxide:acid molar ratio was kept at 1:2. The dissolution of TiO(2) increased with increase in ascorbic acid and oxalic acid concentration up to 0.15M in 4h (corresponding to molar ratio of oxide to acid of 1:3) and further addition did not affect the dissolution. Nearly 45% TiO(2) dissolution was obtained with ascorbic acid alone while oxalic acid yielded 40% dissolution. When oxalic acid was added along with ascorbic acid in equi-molar concentrations, dissolution of TiO(2) was enhanced to 60% in 2.5h but when cysteine was added to ascorbic acid the dissolution was about 50% in just 1h.


Long-term results of tricuspid reconstruction with pericardium in an infant.
We report the long-term clinical outcome of an 8-month-old infant who underwent tricuspid reconstruction using fresh autologous pericardium for severe tricuspid regurgitation due to defects in the leaflets of the tricuspid valve after ventricular septal defect closure and ablation. Ten years after surgery, the tricuspid function is good with mild regurgitation and mild stenosis. From the age of 8 months to 11 years, the patient's body weight and height increased fourfold and twofold, respectively. Thus, a fresh autologous pericardial patch could be the material of choice for tricuspid leaflet repair in pediatric patients.


DR5-associated DC molecules carry three different allospecificities.
Antiserum 8w670 which had been classified as anti-DR5 in the Workshop analysis was found by the direct binding test to react with about 40% of Ia molecules in an Ia preparation from LG38 cells (DR5,5) that was deleted of DR molecules and DR-like molecules we call BR and enriched in DC molecules by pretreatment with a rabbit antiserum raised against alpha-subunits of DR and BR molecules. The specificity involved in the binding reaction was shown by the binding inhibition assay to be present in 100% of DR5-positive cases (53 out of 53) and 8w13-positive cases (2 out of 2) and in 23% of DR4-positive cases (4 out of 17). This association pattern did not correspond to any of the known supertypic specificities including the DC beta 4 and DC alpha 3, both of which had been found on DR5-associated DC molecules. Yet sequential binding analysis revealed that the 8w670-defined specificity was indeed present on the same molecules carrying DC beta 4 and DC alpha 3 specificities. This specificity was designated DC5.


Seizures in the newborn.
Seizures in the newborn period constitute a medical emergency. Subtle seizures are the commonest type of neonatal seizures, other types being clonic, tonic, and myoclonic. Myoclonic seizures carry the worst prognosis in terms of long-term neurodevelopmental outcome. Hypoxic-ischemic encephalopathy is the most common cause of neonatal seizures. Multiple etiologies often co-exist in neonates and hence it is essential to rule out conditions such as hypoglycemia, hypocalcemia, and meningitis before initiating specific therapy. A comprehensive approach for management of neonatal seizures has been described.


Up-regulation of Bak and Bim via JNK downstream pathway in the response to nitric oxide in human glioblastoma cells.
Nitric oxide (NO) is a chemical messenger implicated in neuronal damage associated with ischemia neurodegenerative disease and excitotoxicity. In the present study, we examined the biological effects of NO and its mechanisms in human malignant glioblastoma cells. Addition of a NO donor, S-nitroso-N-acetyl-penicillamine (SNAP), induced apoptosis in U87MG human glioblastoma cells, accompanied by opening mitochondrial permeability transition pores, release of cytochrome c and AIF, and subsequently by caspase activation. NO-induced apoptosis occurred concurrently with significantly increased levels of the Bak and Bim. Treatment with SNAP resulted in sustained activation of JNK and its downstream pathway, c-Jun/AP-1. The expression of dominant-negative (DN)-JNK1 and DN-c-Jun suppressed the activation of AP-1, the induction of Bak and Bim, and the SNAP-induced apoptosis. In addition, de novo protein synthesis was required for the initiation of apoptosis in that the protein synthesis inhibitor, cycloheximide (CHX), inhibited NO-induced apoptotic cell death as well as up-regulation of Bak and Bim. These results suggest that NO activates an apoptotic cascade, involving sustained JNK activation, AP-1 DNA binding activity, and subsequent Bak and Bim induction, followed by cytochrome c and AIF releases and caspases cascade activation, resulting in human malignant brain tumor cell death.


Validation of transpulmonary thermodilution cardiac output measurement in a pediatric animal model.
This study was undertaken to validate the transpulmonary thermodilution cardiac output measurement (CO(TPTD)) in a controlled newborn animal model under various hemodynamic conditions with special emphasis on low cardiac output. Prospective, experimental, pediatric animal study. Animal laboratory of a university hospital. Twelve lambs. We studied 12 lambs under various hemodynamic conditions. Cardiac output was measured using the transpulmonary thermodilution technique with central venous injections of ice-cold saline. An ultrasound transit time perivascular flow probe around the main pulmonary artery served as the standard reference measurement (CO(UFP)). During the experiment, animals were resuscitated from hemodynamic shock using fluid boluses. Cardiac output measurements were performed throughout the experiment. The correlation coefficient between CO(TPTD) and CO(UFP)was .97 (95% confidence interval .94-.98, p < .0001). Bland-Altman analysis showed a mean bias of 0.19 L/min with limits of agreement of -0.04 and 0.43 L/min (12.0% and +/-14.7%, respectively). The correlation coefficient between changes in CO(TPTD) and CO(UFP) during volume loading was .95 (95% confidence interval .91-.96, p < .0001). There was a significant correlation between changes in global end-diastolic volume and changes in stroke volume (r = .59) but not between changes in central venous pressure and changes in stroke volume (r = .03). The transpulmonary thermodilution technique is a reliable method of measuring cardiac output in newborn animals. It is also capable of tracking changes in cardiac output.


Probing rapid carbon fixation in fast-growing seaweed Ulva meridionalis using stable isotope 13C-labelling.
The high growth rate of Ulva seaweeds makes it a potential algal biomass resource. In particular, Ulva meridionalis grows up to fourfold a day. Here, we demonstrated strong carbon fixation by U. meridionalis using 13C stable isotope labelling and traced the 13C flux through sugar metabolites with isotope-ratio mass spectrometry (IR-MS), Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS), 13C-nuclear magnetic resonance spectrometry (13C-NMR), and gas chromatography-mass spectrometry (GC-MS). U. meridionalis was first cultured in 13C-labelled enriched artificial seawater for 0-12 h, and the algae were collected every 4 h. U. meridionalis grew 1.8-fold (dry weight), and the 13C ratio reached 40% in 12 h, whereas 13C incorporation hardly occurred under darkness. At the beginning of the light period, 13C was incorporated into nucleic diphosphate (NDP) sugars in 4 h, and 13C labelled peaks were identified using FT-ICR-MS spectra. Using semiquantitative 13C-NMR measurements and GC-MS, 13C was detected in starch and matrix polysaccharides after the formation of NDP sugars. Moreover, the 14:10 light:dark regime resulted into 85% of 13C labelling was achieved after 72 h of cultivation. The rapid 13C uptake by U. meridionalis shows its strong carbon fixation capacity as a promising seaweed biomass feedstock.


[Successful surgical correction for an incomplete endocardial cushion defect in an elderly patient].
A 72-year-old female who presented with symptoms of severe congestive heart failure, was subsequently diagnosed as having an incomplete endocardial cushion defect. A severe left-to-right shunt and mitral and tricuspid valve regurgitation were noted. Closure of the ostium primum defect, mitral valve replacement, and tricuspid valve annuloplasty were performed simultaneously. The postoperative course was uneventful. A review of literature revealed that this is the oldest such surgical patient described in Japan to date. We believe that surgical correction can be effective even in elderly patients.


In-situ phase transition from microemulsion to liquid crystal with the potential of prolonged parenteral drug delivery.
This study is the first to investigate and demonstrate the potential of microemulsions (MEs) for sustained release parenteral drug delivery, due to phase transition behavior in aqueous environments. Phase diagrams were constructed with Miglyol 812N oil and a blend of (co)surfactants Solutol HS 15 and Span 80 with ethanol. Liquid crystal (LC) and coarse emulsion (CE) regions were found adjacent to the ME region in the water-rich corner of the phase diagram. Two formulations were selected, a LC-forming ME and a CE-forming ME and each were investigated with respect to their rheology, particle size, drug release profiles and particularly, the phase transition behavior. The spreadability in an aqueous environment was determined and release profiles from MEs were generated with gamma-scintigraphy. The CE-forming ME dispersed readily in an aqueous environment, whereas the LC-forming ME remained in a contracted region possibly due to the transition of ME to LC at the water/ME interface. Gamma-scintigraphy showed that the LC-forming ME had minimal spreadability and a slow release of (99m)Tc in the first-order manner, suggesting phase conversion at the interface. In conclusion, owing to the potential of phase transition, LC-forming MEs could be used as extravascular injectable drug delivery vehicles for prolonged drug release.


Impact of environmental factors on the population dynamics, density and foraging activities of Odontotermes lokanandi and Microtermes obesi in Islamabad.
Affect of different environmental factors i.e., temperature, relative humidity and precipitation on population dynamics, density and foraging activities of Microtermes obesi Holmgren and Odontotermes lokanandi Chatarjee and Thakur (Isoptera: Termitidae) were studied from March 2010 to July 2012 in Islamabad. A total of 1200 poplar wooden stakes was used for monitoring the termite activities in Islamabad. The results showed that 65 out of 1200 poplar wooden stakes were found infested by both species i.e. M. obesi and O. lokanandi. Both species were interacting with each other in the experimental field and O. lokanandi was found significantly dominant. Mean yield per trap ranged from 0.83 ± 0.20 gm to 1.12 ± 0.28 gm and 0.35 ± 0.09 gm to 0.82 ± 0.19 gm for M. obesi and O. lokanandi in the field, respectively. M. obesi and O. lokanandi in 1.0 gm sample ranged from 539.83 ± 2.21 to 567.83 ± 9.41 and 407.67 ± 4.75 to 424.5 ± 1.15 individuals, respectively. Population of workers ranged from 93.53 ± 1.73 to 97.68 ± 0.40 and 91.69 ± 1.42 to 98.41 ± 0.50 percent for M. obesi and O. lokanandi, respectively. Positive and significant correlation was found among atmospheric temperature, precipitation and both subterranean termite species i.e., M. obesi and O. lokanandi; however, the correlation was found non significant and negative between relative humidity and foraging activities of both termite species. Moreover, correlation was found positive and significant between atmospheric temperature and percent workers of M. obesi; while negative and non-significant between atmospheric temperature and percent workers of O. lokanandi. Negative and significant correlation was noted between relative humidity and percent workers of M. obesi; whereas, positive and significant correlation was recorded between relative humidity and percent workers of O. lokanandi. Positive and non-significant correlation was recorded between precipitation and percent workers of M. obesi; while positive and significant correlation was observed between precipitation and percent workers of O. lokanand.


Screening cascade and development of potential Positron Emission Tomography radiotracers for mGluR5: in vitro and in vivo characterization.
Use of mGluR5 receptor radiotracers to determine whether an in vitro binding assay is able to predict how good a radiotracer is likely to be in imaging receptor in the central nervous system (CNS) via positron emission tomography (PET). Saturation and equilibrium competition studies in rat and rhesus membranes were used to determine receptor concentrations and tracer affinities. In addition, specific binding of metabotropic receptor subtype 5 (mGluR5) radioligands in rhesus and rat brain sections was determined using a "no-wash protocol," and the in vivo binding signal in rats was determined using micro-PET. Affinity values were determined for a series of mGluR5 antagonists (1-5) and ranged from 0.1 to 11 nM in rat. A previously reported "no-wash protocol" was then employed to determine specific binding in tissue sections following a 20-min incubation, and the regional distribution of these mGluR5 radiotracers determined in rat brain via autoradiography. The analogs 1b, 2b, 3b, and 4b, but not 5b, displayed good signal-to-noise ratios under these conditions with high density of binding in caudate, cortex, and hippocampus and lower density in cerebellum. With this information it was predicted that 1c, 2c, 3b, and 4b would display measurable signal-to-noise ratios in vivo, and that the larger in vitro signals for 3b and 4b would translate to 3b and 4b yielding the best in vivo signals. These predictions were investigated using micro-PET imaging in rat. Compound 1c showed a rapid wash-in and rapid wash-out profile in rat brain. Compound 2c showed similar signal-to-noise ratio as 1b, but slower washout. Compounds 3b and 4b showed the best signal-to-noise ratio in vivo, while 5b did not provide a significant signal, as predicted. In vivo occupancy estimates for 2-methyl-6-(phenylethynyl)-pyridine (MPEP) following intravenous administration were determined using radiolabeled compounds 1c, 2c, and 3b; they were essentially the same and were on the order of 1 mg kg(-1) (ID(50)). An in vitro screen of several mGluR5 tracers was used to rapidly predict whether radiolabeled mGluR5 analogs would be useful as PET radiotracers. Results provided an extension to previously reported data. Two of the four radiotracers with the best in vitro "no-wash" results also showed the best potential as measured noninvasively using micro-PET.


A longitudinal study of falls in an elderly population II. Some circumstances of falling.
Some of the circumstances surrounding 651 falls which occurred in elderly residents of a standard care unit in a home for the aged are described. Falls occurred as frequently at night as during the day-time, and there was no significant variation by day of the week or by season. The great majority occurred inside the home, mainly in the resident's own suite. The subsequent morbidity varied according to the place of occurrence, the percentage of severe injuries being lower in falls occurring in toilets and elevators, and to a lesser extent hallways, compared with those occurring in other inside locations and outside. The commonest activity associated with falling was going to or returning from the toilet. However, one-third of day-time falls occurred when the resident was sitting in a chair. The severity of injury did not vary with associated activity. Possible explanations for these findings are discussed, particularly from the preventive viewpoint.


Keratoconus enlargement as a predictor of keratoconus progression.
Numerous approaches have been designated to document progression in keratoconus, nevertheless there is no consistent or clear definition of ectasia progression. In this present study, we aim to evaluate Keratoconus Enlargement (KCE) as a parameter to document ectasia progression. We define KCE as an increase of more than 1D in the anterior curvature of non-apical corneal areas. We have designed a longitudinal study in 113 keratoconic eyes to assess keratoconus progression. KCE was compared with variables commonly used for detection of keratoconus progression like Kmax, Km, K2, PachyMin, D-Index, Corneal Astigmatism and PRC of 3.0 mm centered on the thinnest point. The variations of keratometric readings, D-index and ELEBmax showed positive associations with KCE. Evaluating the performance of Kmax, D-index and KCE as isolated parameters to document keratoconus progression we found a sensitivity of 49%, 82% and 77% and a specificity of 100%, 95% and 66% to detect keratoconus progression (p < 0.001 for all). This difference in sensitivity can be explained by the changes in keratoconus outside the small area represented by Kmax. The inclusion of KCE should be considered in the evaluation of keratoconus progression in conjunction with other variables to increase the reliability of our clinical evaluation.


Orofacial features and pediatric dentistry in the long-term management of Infantile Pompe Disease children.
Glycogen storage disease type II (GSDII) or Pompe disease is a rare autosomal recessive metabolic disorder that leads to intracellular glycogen storage in many tissues, mainly in skeletal muscle, heart and liver. Facial muscle weakness and altered craniofacial growth are very common in Pompe disease children. In this paper we describe the orofacial features in two children affected by GSDII and illustrate a multidisciplinary approach that involved enzyme replace therapy, non-invasive ventilation (NIV) and pediatric dentistry with 5-year follow-up. Two Infantile Pompe Disease children were examined by a pediatric dentist at the age of 4 and 5 years old respectively. The orofacial examination showed typical facies with similar features: hypotonia of facial and tongue muscles, lip incompetence, narrow palate with reduction in transversal dimension of the upper dental arch, macroglossia, low position of the tip of the tongue, concave profile, Class III malocclusion with hypoplasia of maxillary-malar area and mandibular prognathism. Myofunctional therapy and orthodontic treatment consisted in oral muscle exercises associated to intraoral and extraoral orthodontic devices. NIV facial mask was substituted with a nasal pillow mask in order to avoid external pressure on the mid-face which negatively influences craniofacial growth. This paper evidences that the pediatric dentist plays an important role in craniofacial growth control, oral function rehabilitation and, therefore, in the improvement of the quality of life of Pompe children and their families. Therefore an early pediatric dental evalutation should be included in the multidisciplinary management of children suffering from Infantile Pompe Disease.


Automated treatment planning for proton pencil beam scanning using deep learning dose prediction and dose-mimicking optimization.
The purpose of this study is to investigate the use of a deep learning architecture for automated treatment planning for proton pencil beam scanning (PBS). A 3-dimensional (3D) U-Net model has been implemented in a commercial treatment planning system (TPS) that uses contoured regions of interest (ROI) binary masks as model inputs with a predicted dose distribution as the model output. Predicted dose distributions were converted to deliverable PBS treatment plans using a voxel-wise robust dose mimicking optimization algorithm. This model was leveraged to generate machine learning (ML) optimized plans for patients receiving proton PBS irradiation of the chest wall. Model training was carried out on a retrospective set of 48 previously-treated chest wall patient treatment plans. Model evaluation was carried out by generating ML-optimized plans on a hold-out set of 12 contoured chest wall patient CT datasets from previously treated patients. Clinical goal criteria and gamma analysis were used to compare dose distributions of the ML-optimized plans against the clinically approved plans across the test patients. Statistical analysis of mean clinical goal criteria indicates that compared to the clinical plans, the ML optimization workflow generated robust plans with similar dose to the heart, lungs, and esophagus while achieving superior dosimetric coverage to the PTV chest wall (clinical mean V95 = 97.6% vs. ML mean V95 = 99.1%, p < 0.001) across the 12 test patients. ML-based automated treatment plan optimization using the 3D U-Net model can generate treatment plans of similar clinical quality compared to human-driven optimization.


Dose-dependent lesions of the dopaminergic nigrostriatal pathway induced by intrastriatal injection of 6-hydroxydopamine.
Animal models with partial lesions of the dopaminergic nigrostriatal pathway may be useful for developing neuroprotective and neurotrophic therapies for Parkinson's disease. To develop such a model, different doses of 6-hydroxydopamine (0.0, 0.625, 1.25, 2.5 and 5.0 micrograms/microliters in 3.5 microliters of saline) were unilaterally injected into the striatum of rats. Animals that received 1.25 to 5.0 micrograms/microliters 6-hydroxydopamine displayed dose-dependent amphetamine and apomorphine-induced circling. 6-Hydroxydopamine also caused dose-dependent reductions in [3H]mazindol-labeled dopamine uptake sites in the lesioned striatum and ipsilateral substantia nigra pars compacta (up to 93% versus contralateral binding), with smaller losses in the nucleus accumbens, olfactory tubercle and ventral tegmental area. In the substantia nigra pars compacta and the ventral tegmental area, the number of Nissl-stained neurons decreases in parallel with the reduction in [3H]mazindol binding. The reduction in [3H]mazindol binding in the striatum and the nucleus accumbens, and the reduction in [3H]mazindol binding and in the number of Nissl-stained neurons in the substantia nigra pars compacta and the ventral tegmental area is stable for up to 12 weeks after the lesion. Macroscopically, forebrain coronal sections showed normal morphology, except for rats receiving 5.0 micrograms/microliters 6-hydroxydopamine in which striatal cross-sectional area was reduced, suggesting that this high dose non-specifically damages intrinsic striatal neurons. Nissl-stained sections revealed an area of neuronal loss and intense gliosis centered around the needle track, which increased in size with the dose of neurotoxin. Striatal [3H]sulpiride binding was increased by 2.5 micrograms/microliters and 5.0 micrograms/microliters 6-hydroxydopamine, suggesting up-regulation of dopamine D2 receptors. Striatal binding of [3H]CGS 21680-labeled adenosine A2a receptors, but not of [3H]SCH 23390-labeled dopamine D1 receptors, was reduced at the highest dose, suggesting preservation of the striatal intrinsic neurons with the lower doses. This study indicates that intrastriatal injection of different doses of 6-hydroxydopamine can be used to cause increasing amounts of dopamine denervation, which could model Parkinson's disease of varying degrees of severity. Injecting 3.5 microliters of 2.5 micrograms/microliters 6-hydroxydopamine appears to be particularly useful as a general model of early Parkinson's disease, since it induces a lesion characterized by robust drug-induced rotation, changes in binding consistent with approximately 70% dopamine denervation, approximately 19% dopamine D22 receptor up-regulation, negligible intrinsic striatal damage and stability for at least 12 weeks. This study outlines a technique for inducing partial lesions of the nigrostriatal dopamine pathway in rats.(ABSTRACT TRUNCATED AT 400 WORDS)


Production of human β-actin and a mutant using a bacterial expression system with a cold shock vector.
Actin is the most abundant protein in the cytoplasm of most eukaryotic cells and is involved in a variety of cellular functions. It has been difficult to produce actin in bacterial expression systems in good yields. In this study, we developed a new simple method for the production of recombinant actin in Escherichia coli cells. Human β-actin was successfully expressed using a cold shock vector, pCold, in the bacterial expression system. The expressed β-actin (hexahistidine-tagged) was separated with a Ni-chelating resin, followed by a polymerization/depolymerization cycle or column chromatography with the Ni-chelating resin. The purified recombinant β-actin showed a normal polymerization ability compared with β-actin purified from human platelets. We produced a recombinant mutant actin with a Gly-168Arg mutation in the system and confirmed that it exhibited an impaired polymerization ability. The system developed in this study will provide a useful method for the production of actin isoforms and their mutants.


Extent of resection and survival outcomes in World Health Organization grade II meningiomas.
WHO grade II meningiomas behave aggressively, with recurrence rates as high as 60%. Although complete resection in low-grade meningiomas is associated with a relatively low recurrence rate, the impact of complete resection for WHO grade II meningiomas is less clear. We studied the association of extent of resection with overall and progression-free survivals in patients with WHO grade II meningiomas. A retrospective database review was performed to identify all patients who underwent surgical resection for intracranial WHO grade II meningiomas at our institution between 1995 and 2019. Kaplan-Meier analysis was used to compare overall and progression-free survivals between patients who underwent gross total resection (GTR) and those who underwent subtotal resection (STR). Multivariable Cox proportional-hazards analysis was used to identify independent predictors of tumor recurrence and mortality. Of 214 patients who underwent surgical resection for WHO grade II meningiomas (median follow-up 53.4 months), 158 had GTR and 56 had STR. In Kaplan-Meier analysis, patients who underwent GTR had significantly longer progression-free (p = 0.002) and overall (p = 0.006) survivals than those who underwent STR. In multivariable Cox proportional-hazards analysis, GTR independently predicted prolonged progression-free (HR 0.57, p = 0.038) and overall (HR 0.44, p = 0.017) survivals when controlling for age, tumor location, and adjuvant radiation. Extent of resection independently predicts progression-free and overall survivals in patients with WHO grade II meningiomas. In an era of increasing support for adjuvant treatment modalities in the management of meningiomas, our data support maximal safe resection as the primary goal in treatment of these patients.


Ensemble classifier fostered detection of arrhythmia using ECG data.
Electrocardiogram (ECG) is a non-invasive medical tool that divulges the rhythm and function of the human heart. This is broadly employed in heart disease detection including arrhythmia. Arrhythmia is a general term for abnormal heart rhythms that can be identified and classified into many categories. Automatic ECG analysis is provided by arrhythmia categorization in cardiac patient monitoring systems. It aids cardiologists to diagnose the ECG signal. In this work, an Ensemble classifier is proposed for accurate arrhythmia detection using ECG Signal. Input data are taken from the MIT-BIH arrhythmia dataset. Then the input data was pre-processed using Python in Jupyter Notebook which run the code in an isolated manner and was able to keep code, formula, comments, and images. Then, Residual Exemplars Local Binary Pattern is applied for extracting statistical features. The extracted features are given to ensemble classifiers, like Support vector machines (SVM), Naive Bayes (NB), and random forest (RF) for classifying the arrhythmia as normal (N), supraventricular ectopic beat (S), ventricular ectopic beat (V), fusion beat (F), and unknown beat (Q). The proposed AD-Ensemble SVM-NB-RF method is implemented in Python. The proposed AD-Ensemble SVM-NB-RF method is 44.57%, 52.41%, and 29.49% higher accuracy; 2.01%, 3.33%, and 3.19% higher area under the curve (AUC); and 21.52%, 23.05%, and 12.68% better F-Measure compared with existing models, like multi-model depending on the ensemble of deep learning for ECG heartbeats arrhythmia categorization (AD-Ensemble CNN-LSTM-RRHOS), ECG signal categorization utilizing VGGNet: a neural network based classification method (AD-Ensemble CNN-LSTM) and higher performance arrhythmic heartbeat categorization utilizing ensemble learning along PSD based feature extraction method (AD-Ensemble MLP-NB-RF).


Increased osteopontin levels in the cerebrospinal fluid of patients with multiple sclerosis.
To determine cerebrospinal fluid levels of osteopontin (OPN), a proinflammatory cytokine that was found to be overexpressed in multiple sclerosis lesions and increased in plasma during relapses and in secondary progressive multiple sclerosis. Case series. Osteopontin, interleukin 12p40 (IL-12p40), IL-10, and matrix metalloproteinase 9 were measured by enzyme-linked immunosorbent assay by an investigator unaware of the patients' diagnoses. Consecutive patients with multiple sclerosis (n = 27), or other inflammatory (n = 11) or non-inflammatory (n = 23) neurological diseases, undergoing lumbar puncture, were investigated. Osteopontin was significantly elevated in the cerebrospinal fluid of patients with multiple sclerosis (mean [SD], 415 [186] ng/mL) and other inflammatory diseases (563 [411] ng/mL) compared with those with noninflammatory neurological diseases (286 [150] ng/mL). Cerebrospinal fluid OPN levels were slightly higher than plasma OPN levels. Cerebrospinal fluid OPN levels positively correlated with the ability to detect cerebrospinal fluid IL-12p40. Osteopontin in the cerebrospinal fluid may be, in part, of central nervous system origin, and may play an important role in central nervous system inflammation.