Results of endoscopic management of carpal tunnel syndrome.
This paper is a retrospective study of 27 women with carpal tunnel syndrome (39 hands) who underwent a new endoscopic operative procedure utilizing the Universal Subcutaneous Endoscope system developed by the lead author. Operations on 199 hands were performed under local anesthesia on an outpatient basis. The etiology was considered idiopathic in all cases. Complete preoperative and postoperative clinical and electrophysiologic data were analyzed in 39 hands. The follow-up period ranged from 12 to 45.8 months (mean, 18.3 months). Symptoms of sensory disturbances disappeared in an average of 20 weeks in all patients. Electrophysiologic studies showed definite improvement when compared with preoperative studies. No complications were recorded. Clinical and electrophysiologic results showed that the less-invasive management of carpal tunnel syndrome by endoscopy is safe and effective.


Beta 1-selective and non-selective beta-adrenoceptor blockade, anaerobic threshold and respiratory gas exchange during exercise.
The effect of oral doses of the beta 1-selective adrenoceptor antagonist atenolol (50 mg), the non-selective antagonist propranolol (40 mg) and placebo was investigated during exercise in a crossover comparison in six healthy but untrained subjects. Descriptors of ventilation, respiratory gas exchange, and arterialized blood lactate and glucose were obtained during steady state bicycle ergometric exercise at 20% and 60% of the subjects' previously determined maximal oxygen uptake (VO2 max). At these work intensities, the previously reported increase of respiratory exchange ratio (RER) during non-selective beta-adrenoceptor blockade was found to be trivial (placebo = 0.96 +/- 0.03 s.e. mean; propranolol = 0.97 +/- 0.01; atenolol = 0.97 +/- 0.04; 60% VO2 max, 10 min exercise) and only present during the early minutes of effort. Oxygen uptake and carbon dioxide production did not differ between treatments. Both drugs produced highly significant falls in peak expiratory flow (PEF) rates and tidal volume (VT) which were compensated by an increase in respiratory rate. PEF, 60% VO2 max: placebo = 3.8 +/- 0.3 l/s; propranolol 3.6 +/- 0.3 l/s (P less than 0.03); atenolol 3.1 +/- 0.3 l/s (P less than 0.01). VT, 60% VO2 max: placebo 2.0 +/- 0.1 l; propranolol 1.8 +/- 0.21 (P less than 0.05); atenolol 1.7 +/- 0.1 1 (P less than 0.01). Arterialized lactate was significantly elevated during work at 20% and 60% VO2 max, but rose progressively at the 60% VO2 max load. Ventilation, oxygen uptake and ventilatory equivalent for carbon dioxide also rose progressively at this workload. Ventilatory equivalent for oxygen showed no significant rise.(ABSTRACT TRUNCATED AT 250 WORDS)


Physician numeracy as the basis for an evidence-based medicine curriculum.
Most medical schools and postgraduate programs devote some time to teaching evidence-based medicine (EBM). EBM encompasses five essential skills, including constructing a sound clinical question, literature searching, critical appraisal, gaining a full understanding of study results, and integration of results into patient care. Gaining a full understanding of results requires understanding the statistical aspects of and terminology associated with the design, analysis, and results of original research--hereby referred to as physician numeracy. Physicians and physicians-in-training recognize the importance of these concepts but are uncomfortable with and demonstrate poor knowledge of the quantitative aspects of research. This is not surprising since few curricula include physician numeracy. Current approaches to teaching EBM rely on journal clubs, which have not been shown to improve participants' self-perceived EBM skills. In this paper we describe a novel approach to teaching EBM which makes use of five guiding principles: (1) Journal clubs have important limitations, (2) understanding the quantitative aspects of research promotes an in-depth understanding of papers, (3) physician numeracy can form the basis of an EBM course, (4) consumers of original research ought to determine what is useful about a paper rather than whether or not it is useful, and (5) numeracy should encompass only those concepts needed to accurately interpret evidence and apply it to individual patients. An EBM curriculum based on physician numeracy is described, together with the challenges inherent to this approach.


Photooxidation of specific residues in alpha-crystallin polypeptides.
Singlet oxygen is a biologically important, photochemically generated species that preferentially oxidizes His, Trp, and Met residues of protein molecules. Calf alpha-crystallin was photooxidized with use of meso-tetra(p-sulfonatophenyl)porphyrin (TPPS) and uroporphyrin (UP) as singlet oxygen generators. The effects of photooxidation were monitored by analyzing the changes in alpha-crystallin peptide maps obtained by reversed-phase HPLC using a photodiode array absorbance detector. The reaction led to the loss of six specific peptides, five of which contained photooxidizable residues. Peptides containing His-97 and His-154 from the A chain and Met-68 from the B chain are preferentially photooxidized, suggesting that those residues have access to singlet oxygen. Trp residues in the N-terminal region are converted to NFK, whereas Trp-60 in the B chain is not photooxidized strongly suggesting that the former are close to the surface of alpha-crystallin while the latter Trp residue is buried. Only one peptide that is lost from the peptide maps does not contain a photooxidizable group; however, this peptide does contain an apparently undigested Lys residue. It is suggested that it forms a cross-link with a photooxidized His residue.


The household structure of U.S. immigrants and natives.
The household structures of U.S. immigrants and natives are compared in an attempt to assess migrant assimilation. Data from the 1976 Survey of Income and Education are used to "compare actual household structures of immigrants by country of origin to those of natives. Linear probability regression models are estimated to determine whether and to what extent the household structure of immigrants [converges] to that of natives as the number of years that the immigrant has lived in the United States approaches the total years of life....[It is found that] previous immigrants to the United States have adopted in their living arrangements forms of household structure that appear to converge toward those of natives with time in the United States."


Variation in particle shape of active pharmaceutical ingredients prepared by fluidized-bed jet-milling.
In pharmaceutical industries, most active pharmaceutical ingredients are poorly water soluble, and therefore milling processes are important to obtain fine particles that can be easily dissolved in the body. However, the main purpose of milling is micronization of particles. From the viewpoint of fine particle preparation in the formulation process, milling has not been investigated sufficiently. In this paper, ethenzamide was milled under various operating conditions using a fluidized-bed jet-mill. It was found that not only the particle size but also the particle shape varied with the milling conditions. The relationship between particle shape and milling conditions has been obtained experimentally.


Interface engineering strategy for multisource spintronic devices via TMPS4 modulation of black-phosphorus.
Interface engineering is an effective strategy for significantly providing performance improvements in logic operations and information storage techniques, given the quantum effects characteristic of pristine two-dimensional ferromagnetic (FM) materials. Furthermore, the van der Waals (vdW) heterostructures enable a more efficient design of logic components. Herein, two novel designs of van der Waals heterostructures are proposed, black-phosphorus (BP)/TMPS4(TM = Cr, Fe-Mn), where the CrPS4 relies on odd and even layers for FM state - antiferromagnetic (AFM) state alternation, which is used to modulate BP with high hole mobility but no magnetism. Based on the first principles, the simulation results show that the two heterojunctions exhibit high stability, carrier mobility, and thermoelectric effects, of which the BP/CrPS4 heterojunction is specifically modulated to a type II electronic bandstructure. These two structures are applied in multi-source logic devices. The device performances show that the devices have spin Sebeck effect (SSE), perfect spin filtering effect (SFE), high extinction ratio (1347), and high thermal magnetoresistivity (1011). The above results suggest BP/TMPS4 bilayers as promising candidates for spin-based vdW devices and facilitate the future development of atomically thin magnetic information storage.


Evaluation of antibacterial and antifungal compounds for selective inhibition of denitrification in soils.
Nitrous oxide (N2O) is an atmospheric constituent implicated in climate warming and stratospheric ozone depletion. Both bacteria and fungi participate in N2O production, but information is lacking with regard to the relative contribution of bacterial and fungal denitrifiers to the denitrification process in agricultural soils. The selective inhibition (SI) technique is widely used to assess the contribution of different groups of microbes to soil processes, but success of the technique depends on the effectiveness of the inhibitors. In this study, laboratory experiments were conducted to assess the contribution of bacteria and fungi to denitrification using soils from a woodlot, agricultural fields under conventional plowing (PT), and no-till for either 50 years (long-term) or 11 years (medium-term). A selective inhibition (SI) technique was developed using two bactericides (streptomycin and bronopol) and two fungicides (cycloheximide and captan) applied at different rates (0-32 mg per g soil). Regardless of the application rate, streptomycin and cycloheximide were not effective inhibitors of denitrification, with a degree of inhibition only between 2 and 20% relative to controls. These results are significant given the wide use of these products in SI studies. However, the bactericide bronopol and the fungicide captan effectively inhibited denitrification, with the strongest inhibition observed at an application rate of 16 mg per g soil. The ratio of fungal to bacterial denitrification activity (F : B) was generally less than 1, indicating a dominance of bacteria in denitrification activity in the soils investigated. However, an increase in the F : B ratio from 0.24 in medium-term NT to 0.87 in long-term NT soils was noted, suggesting perhaps a progressive increase in the role of fungal denitrifiers with a longer duration of NT farming.


Marked decrease of neuropeptide Y Y2 receptor binding sites in the hippocampus in murine prion disease.
Using autoradiographic binding methodology with monoiodinated peptide YY together with the agonists neuropeptide Y (NPY) and NPY (13-36), as well as in situ hybridization with oligonucleotide probes complementary to the NPY Y2 receptor (Y2-R) mRNA, we have studied whether or not intracerebral prion inoculation affects Y2-Rs in male CD-1 mice. Monoiodinated peptide YY binding, mainly representing Y2-Rs, was down-regulated by 85% in the CA1 strata oriens and radiatum and by 50-65% in the CA3 stratum oriens 110-140 days postinoculation. In the CA3 stratum radiatum, where the mossy fibers from the dentate granule cells project, there was a significant decrease in PYY binding at 110-120 days. Y2-R mRNA, moderately expressed both in the CA1 and CA3 pyramidal cell layers and the granule cell layer in the dentate gyrus, showed a slight, but not significant, decrease in CA3 neurons 130 days postinoculation. The results indicate that the accumulation of the scrapie prion protein in the CA1-3 region strongly inhibits NPY binding at the Y2-Rs, which, however, is only marginally due to reduced Y2-R mRNA expression. The loss of the ability of NPY to bind to inhibitory Y2-Rs may cause dysfunction of hippocampal circuits and may contribute to the clinical symptoms in mouse scrapie.


Induction chemotherapy followed by radiotherapy in the treatment of anal cancer.
This study was conducted to investigate the efficacy of the induction chemotherapy followed by radiotherapy in anal cancer. Twenty-three patients diagnosed with anal cancer between March 1991 and February 1999 were treated with induction chemotherapy with 5-fluorouracil based regimens followed by external beam radiation. After a median follow-up of 78 months, the overall survival and the disease-free survival at 5 years was 71.3 and 67.5%, respectively. The colostomy-free survival at 5 years was 91%, as the inguinal lymph nodes were the most frequent site of relapse. Serious side effects did not occur and late complications did not develop either. Induction chemotherapy followed by radiotherapy in patients with anal cancer is effective in terms of its response and preserving the anal sphincter without serious acute and late complications.


The Parathyroid Hormone-Related Protein/Parathyroid Hormone 1 Receptor Axis in Adipose Tissue.
Adipose tissue is an organ that shows great plasticity and is able to adapt to the conditions to which the body is subdued. It participates in the regulation of energetic homeostasis and has endocrine functions. Recent studies have shown how the parathyroid hormone-related protein (PTHrP)/Parathyroid Hormone Receptor 1 (PTH1R) axis participates in the regulation of adipogenesis, opposing the action of Peroxisome proliferator-activated receptor gamma (PPARγ). In addition to this, PTHrP is overexpressed in adipose tissue in situations of wear and tear of the body, favoring browning and lipolysis in this tissue. It is also overexpressed in adipose tissue in stressful situations but in the opposite direction, in obesity, metabolic syndrome, type 2 diabetes mellitus (T2DM) and gestational diabetes mellitus (GDM). In conclusion, the PTHrP/PTH1R axis has a main role in adipose tissue, participating in its differentiation and remodeling. PTHrP might be used in obesity treatment and its complications for its ability to reprogram adipogenesis and adipose tissue expansion, WAT browning and for the improvement of the insulin sensitivity. In addition, PTHrP could even be used as a marker of placental status and maternal adaptations to prevent future metabolic problems in mothers and children, as well as in the treatment of bone-related diseases such as osteoporosis.


A mechanogated nonselective cation channel in proximal tubule that is ATP sensitive.
Ion channels that are gated in response to membrane deformation or "stretch" are empirically designated stretch-activated channels. Here we describe a stretch-activated nonselective cation channel in the basolateral membrane (BLM) of the proximal tubule (PT) that is nucleotide sensitive. Single channels were studied in cell-intact and cell-free patches from the BLM of PT cells that maintain their epithelial polarity. The limiting inward Cs+ conductance is ~28 pS, and channel activity persists after excision into a Ca2+- and ATP-free bath. The stretch-dose response is sigmoidal, with half-maximal activation of about -19 mmHg at -40 mV, and the channel is activated by depolarization. The inward conductance sequence is: NH ~ Cs+ ~ Rb+ > K+ ~ Na+ ~ Li+ > Ca2+ ~ Ba2+ > N-methyl-D-glucamine ~ tetraethylammonium. The venom of the common Chilean tarantula, Grammostola spatulata, completely blocks channel activity in cell-attached patches. Hypotonic swelling reversibly activates the channel. Intracellular ATP concentration ([ATP]i) reversibly blocks the channel (inhibitory constant approximately 0.48 mM), suggesting that channel function is coupled to the metabolic state of the cell. We conclude that this channel may function as a Ca2+ entry pathway and/or be involved in regulation of cell volume. We speculate this channel may be important when [ATP]i is depleted, as occurs during periods of increased transepithelial transport or with ischemic injury.


[Effects of periodontitis patient's own tissue nucleic acid on the mRNA expression of osteoclast-related factors in murine macrophages].
This paper aimed to determine the mRNA expression of osteoclast-related factors interleukin-6 (IL-6), interleukin-12 (IL-12) p35, IL-12p40, matrix metalloproteinase-9 (MMP-9), nuclear factor of activated T-cells cytoplasmic 1 (NFATcl), receptor activator of nuclear factor-KB (RANK), and tumor necrosis factor-α (TNF-α) mRNA in murine macrophages infected by a periodontitis patient's own tissue nucleic acid. Another aim was to investigate the effects of a periodontitis patient's own tissue nucleic acid on the differentiation of macrophages into osteoclasts. Inflammatory periodontal tissue samples of chronic periodontitis patients were taken during periodontal flap surgery, and healthy gingival tissue samples were taken from orthodontic patients during tooth extractions. Total RNA from periodontal tissue was extracted and reversely transcribed into cDNA and then cryo-preserved until further use. First, specific sequence oligodeoxynucleotide MT0I at a concentration of 1 µg · mL⁻¹ was added in murine macrophage RAW264.7, and the cells were incubated for 3 hours. Cells with PBS (1 µg · mL⁻¹) were used as negative controls. The inflammatory periodontal tissue cDNA and healthy periodontal tissue cDNA (1 µg · mL⁻¹) was added subsequently. There were four experimental groups: healthy periodontal tissue cDNA+ RAW264.7, inflammatory periodontal tissue cDNA+RAW264.7, MT01+healthy periodontal tissue cDNA+RAW264.7, and MT01+inflammatory periodontal tissue cDNA+RAW264.7. Real-time quantitative polymerase chain reaction was used to detect the mRNA expression of osteoclast-related factors IL-6, IL-12p35, IL-12p4O, MMP-9, NFATcl, RANK, and TNF-α mRNA after 3, 6, 12, and 24-hours. The mRNA levels of osteoclast-related factors NFATc1, MMP-9, TNF-a, IL-6, IL-12p40, IL-12p35, and RANK in RAW264.7 were markedly upregulated with the treatment of periodontitis patient's own tissue nucleic acid. However, the mRNA expression of osteoclast-related factors was inhibited by use of an immunosuppressant MT01. The periodontitis patient's own tissue nucleic acid could promote the differentiation of murine macrophage into osteoclasts.


Adipose Stem Cells in Regenerative Medicine: Looking Forward.
Over the last decade, stem cell-based regenerative medicine has progressed to clinical testing and therapeutic applications. The applications range from infusions of autologous and allogeneic stem cells to stem cell-derived products. Adult stem cells from adipose tissue (ASCs) show significant promise in treating autoimmune and neurodegenerative diseases, vascular and metabolic diseases, bone and cartilage regeneration and wound defects. The regenerative capabilities of ASCs in vivo are primarily orchestrated by their secretome of paracrine factors and cell-matrix interactions. More recent developments are focused on creating more complex structures such as 3D organoids, tissue elements and eventually fully functional tissues and organs to replace or repair diseased or damaged tissues. The current and future applications for ASCs in regenerative medicine are discussed here.


HMGB1 Mediates Autophagy Dysfunction via Perturbing Beclin1-Vps34 Complex in Dopaminergic Cell Model.
Parkinson's disease (PD), a progressive neurodegenerative disorder, is characterized by irreversible dopaminergic neuron loss and intra-neuronal α-synuclein aggregation. High mobility group box 1 (HMGB1) has been proven to be involved in autophagy dysfunction induced by α-synuclein accumulation, and the Beclin1-vacuolar protein sorting 34 (Vps34) complex is of great importance to the initiation of autophagy. Nevertheless, the concrete interaction mechanism between HMGB1, α-synuclein and autophagy remains elusive, especially in the context of PD. Here in this study, we investigated the interaction between HMGB1 and α-synuclein in rotenone-induced PD cell models and their roles in autophagy flux. Results revealed elevated expression and cytosolic translocation of endogenous HMGB1 upon rotenone exposure. Besides, HMGB1 was found to be able to co-localize and interact with α-synuclein. Moreover, it had also been proven that HMGB1 could aggravate α-synuclein aggregation induced autophagy dysfunction via perturbing Beclin1-Vps34 complex formation. Based on these findings, we propose that HMGB1 is involved in rotenone-induced dopaminergic cell death via interacting with α-synuclein, perturbing the autophagy process, aggravating protein aggregation and finally propelling dopaminergic neurons to move from morbidity to mortality.


Crystallization and preliminary X-ray analysis of an alanine dehydrogenase from Bacillus megaterium WSH-002.
Alanine dehydrogenase (L-AlaDH) from Bacillus megaterium WSH-002 catalyses the NAD⁺-dependent interconversion of L-alanine and pyruvate. The enzyme was expressed in Escherichia coli BL21 (DE3) cells and purified with a His6 tag by Ni²⁺-chelating affinity chromatography for X-ray crystallographic analysis. Crystals were grown in a solution consisting of 0.1 M HEPES pH 8.0, 12%(w/v) polyethylene glycol 8000, 8%(v/v) ethylene glycol at a concentration of 15 mg ml⁻¹ purified protein. The crystal diffracted to 2.35 Å resolution and belonged to the trigonal space group R32, with unit-cell parameters a = b = 125.918, c = 144.698 Å.


A review of the curing mechanics of composites and their significance in dental applications.
Much of the technique sensitivity associated with polymer matrix composites is a direct result of their curing shrinkage. Challenges with marginal integrity, adaptation of proximal contact, and residual stress are related to this intrinsic property. There are many test methods described in the literature that measure various aspects of polymerization contraction. Some measure total contraction, which is the sum of pre- and postgelation shrinkage, whereas others are sensitive only to postgelation deformation, which occurs after the onset of measurable stiffness. Development of methods to compensate for curing shrinkage is best described on the basis of an understanding of the polymerization mechanics. The distinction between total and postgelation contraction, and recognition of limitations of test methods are important considerations when interpreting literature data before selecting a restorative material.


Putative role of chloroquine in gene transfer into a human hepatoma cell line by DNA/lactosylated polylysine complexes.
Chloroquine improves drastically the transfection of cells upon exposure to plasmid DNA/glycosylated polylysine complexes. So far the mechanism of action of chloroquine is not well understood. In this paper, the effect of chloroquine was investigated by measuring the transfection efficiency of a human hepatocarcinoma (HepG2 cells) by pSV2LUC/lactosylated polylysine complexes involving their internalization via the galactose-specific membrane lectin of these cells. The luciferase activity in the transfected cells was maximal when the transfection was performed for 3 or 4 h in the presence of 100 microM chloroquine. The luciferase activity was also enhanced in the presence of primaquine, a chloroquine analogue, but was not increased when transfection was performed in the presence of ammonium chloride, methylamine, spermine, or monensin, compounds known to neutralize the pH of the endocytotic vesicle lumen as chloroquine does. Chloroquine enters cells and accumulates in vesicular compartments; the overall intracellular concentration increases to 9 mM, which means that in the vesicular compartment, the chloroquine concentration is still higher. At such high concentrations, chloroquine induces the dissociation of plasmid DNA/lactosylated polylysine complexes, as shown in acellular experiments.


The relationship between obesity, hemoglobin A1c and the severity of COVID-19 at an urban tertiary care center in New York City: a retrospective cohort study.
To determine if obesity and diabetes are risk factors for severe outcomes in COVID-19 and to compare patient outcomes in those two conditions. Retrospective cohort study. Urban tertiary care center in New York City. 302 patients admitted in an inpatient setting, ≥18 years old, with a laboratory-confirmed diagnosis of COVID-19 via nasal PCR swab were randomly selected. Patients were separated into two cohorts based on their body mass index and hemoglobin A1c. 150 patients were placed in the non-obese, non-diabetic cohort and 152 patients were placed in the corresponding cohort (obesity alone, obesity and diabetes, and diabetes alone). Primary outcomes were development of acute kidney injury, commencement of renal replacement therapy, aminotransferase elevation, troponin elevation, lactic acidosis, development of septic shock, use of vasopressors, presence of acute respiratory distress syndrome (ARDS) and intubation. The secondary outcomes were length of stay in days and mortality. Patients with obesity and/or diabetes were more likely to develop ARDS (79 patients vs 57 patients, p<0.0001) and to be intubated (71 patients vs 45 patients, p=0.0031). Patients with obesity and/or diabetes were more likely to require vasopressors (60 patients vs 41 patients, p=0.0284) and to develop lactic acidosis (median 3.15 mmol/L, IQR 1.8 to 5.2 mmol/L, p=0.0432). When comparing patients with diabetes with and without obesity against patients with obesity alone, they were more likely to develop ARDS (87.5%, p=0.0305). Despite these findings, there was no difference in mortality. In patients hospitalised with COVID-19, those with obesity and/or diabetes were more likely to suffer severe complications, but had negligible differences in mortality. This highlights the importance of close monitoring of patients with these conditions and additional areas of research needed to explain the mortality findings.


The Role of HDACs and HDACi in Cartilage and Osteoarthritis.
Epigenetics plays an important role in the pathogenesis and treatment of osteoarthritis (OA). In recent decades, HDAC family members have been associated with OA. This paper aims to describe the different role of HDACs in the pathogenesis of OA through interaction with microRNAs and the regulation of relevant signaling pathways. We found that HDACs are involved in cartilage and chondrocyte development but also play a crucial role in OA. However, the distinct HDAC mechanism in the pathogenesis and treatment of OA require further investigation. Furthermore, HDAC inhibitors (HDACi) can protect cartilage from disease, which may represent a potential therapeutic approach against OA.


Gender differences in dietary intakes: what is the contribution of motivational variables?
Differences between men and women with respect to dietary intakes and eating behaviours have been reported and could be explained by gender differences in motivational variables associated with the regulation of food intake. The main objectives of the present study were to identify gender differences in dietary intakes, eating behaviours and motivational variables and to determine how motivational variables were associated with dietary intakes and eating behaviours in men and women. Sixty-four men and 59 premenopausal women were included in the present study and presented cardiovascular risk factors. The Regulation of Eating Behaviours scale was completed to assess motivational variables. A validated food frequency questionnaire was administered to evaluate dietary intakes and subjects completed the Three-Factor Eating questionnaire to assess eating behaviours. Men had higher energy intake, energy density and percentage of energy from lipids and lower percentage of energy from carbohydrates than women (P ≤ 0.04). Men also had a lower emotional susceptibility to disinhibition than women (P = 0.0001). Women reported a higher score for eating-related self-determined motivation [i.e., eating-related self-determination index (SDI)] than men (P = 0.002). The most notable gender difference in the pattern of associations was that eating-related SDI was negatively associated with energy density (r = -0.30; P = 0.02), only in women. Women had a better dietary profile and higher eating-related SDI than men. However, gender differences in dietary variables might be explained by a potential gender-specific pattern of association of eating-related SDI with dietary intakes and eating behaviours.


Distinct presynaptic and postsynaptic dismantling processes of Drosophila neuromuscular junctions during metamorphosis.
Synapse remodeling is a widespread and fundamental process that underlies the formation of neuronal circuitry during development and in adaptation to physiological and/or environmental changes. However, the mechanisms of synapse remodeling are poorly understood. Synapses at the neuromuscular junction (NMJ) in Drosophila larvae undergo dramatic and extensive remodeling during metamorphosis to generate adult-specific synapses. To explore the molecular and cellular processes of synapse elimination, we performed confocal microscopy, live imaging, and electron microscopy (EM) of NMJ synapses during the early stages of metamorphosis in Drosophila in which the expressions of selected genes were genetically altered. We report that the localization of the postsynaptic scaffold protein Disc large (Dlg) becomes diffuse first and then undetectable, as larval muscles undergo histolysis, whereas presynaptic vesicles aggregate and are retrogradely transported along axons in synchrony with the formation of filopodia-like structures along NMJ elaborations and retraction of the presynaptic plasma membrane. EM revealed that the postsynaptic subsynaptic reticulum vacuolizes in the early stages of synapse dismantling concomitant with diffuse localization of Dlg. Ecdysone is the major hormone that drives metamorphosis. Blockade of the ecdysone signaling specifically in presynaptic neurons by expression of a dominant-negative form of ecdysone receptors delayed presynaptic but not postsynaptic dismantling. However, inhibition of ecdysone signaling, as well as ubiquitination pathway or apoptosis specifically in postsynaptic muscles, arrested both presynaptic and postsynaptic dismantling. These results demonstrate that presynaptic and postsynaptic dismantling takes place through different mechanisms and that the postsynaptic side plays an instructive role in synapse dismantling.


Two weeks of intermittent dobutamine therapy restores cardiac performance and inotropic responsiveness in conscious rats with heart failure.
Dobutamine is frequently used for acute therapy in heart failure. In the present study, the hemodynamic effects of long-term intermittent dobutamine therapy were investigated in conscious rats with heart failure. Rats with healed myocardial infarctions received two i.p. injections of dobutamine per day for 2 weeks. Hemodynamic measurements were performed 90-180 min after the last injection. Two weeks of intermittent dobutamine significantly restored all hemodynamic changes induced by infarction. The maximal cardiac output during volume loading was depressed due to infarction and dose-dependently restored by 2 weeks of intermittent dobutamine. An increased stroke volume accounted for this improvement since the heart rate was not altered. In order to investigate changes in adrenergic responsiveness, the effects of acute dobutamine in nontreated and 2 weeks of dobutamine-treated infarcted rats were compared to those in control rats. Whereas chronotropic responses to acute dobutamine were comparable for all experimental groups, the inotropic response was reduced in nontreated infarcted rats but dose-dependently restored after 2 weeks of intermittent dobutamine therapy. From the data, we conclude that 2 weeks of intermittent dobutamine therapy in conscious rats with healed myocardial infarcts improved cardiac performance and restored the inotropic response to acute dobutamine administration. Data indicate that dobutamine has a long-term effect on cardiac function, which differs from the acute inotropic effect.


DNA, RNA, and protein extraction: the past and the present.
Extraction of DNA, RNA, and protein is the basic method used in molecular biology. These biomolecules can be isolated from any biological material for subsequent downstream processes, analytical, or preparative purposes. In the past, the process of extraction and purification of nucleic acids used to be complicated, time-consuming, labor-intensive, and limited in terms of overall throughput. Currently, there are many specialized methods that can be used to extract pure biomolecules, such as solution-based and column-based protocols. Manual method has certainly come a long way over time with various commercial offerings which included complete kits containing most of the components needed to isolate nucleic acid, but most of them require repeated centrifugation steps, followed by removal of supernatants depending on the type of specimen and additional mechanical treatment. Automated systems designed for medium-to-large laboratories have grown in demand over recent years. It is an alternative to labor-intensive manual methods. The technology should allow a high throughput of samples; the yield, purity, reproducibility, and scalability of the biomolecules as well as the speed, accuracy, and reliability of the assay should be maximal, while minimizing the risk of cross-contamination.


Comparative analysis of drought-responsive transcriptome in Indica rice genotypes with contrasting drought tolerance.
Genetic improvement in drought tolerance in rice is the key to save water for sustainable agriculture. Drought tolerance is a complex trait and involves interplay of a vast array of genes. Several genotypes of rice have evolved features that impart tolerance to drought and other abiotic stresses. Comparative analysis of drought stress-responsive transcriptome between drought-tolerant (DT) landraces/genotypes and drought-sensitive modern rice cultivars will unravel novel genetic regulatory mechanisms involved in stress tolerance. Here, we report transcriptome analysis in a highly DT rice landrace, Nagina 22 (N22), versus a high-yielding but drought-susceptible rice variety IR64. Both genotypes exhibited a diverse global transcriptional response under normal and drought conditions. Gene ontology (GO) analysis suggested that drought tolerance of N22 was attributable to the enhanced expression of several enzyme-encoding genes. Drought susceptibility of IR64 was attributable to significant down-regulation of regulatory components that confer drought tolerance. Pathway analysis unravelled significant up-regulation of several components of carbon fixation, glycolysis/gluconeogenesis and flavonoid biosynthesis and down-regulation of starch and sucrose metabolism in both the cultivars under drought. However, significant up-regulation of α-linolenic acid metabolic pathway observed in N22 under drought appears to be in good agreement with high drought tolerance of this genotype. Consensus cis-motif profiling of drought-induced co-expressed genes led to the identification of novel cis-motifs. Taken together, the results of the comparative transcriptome analysis led to the identification of specific genotype-dependent genes responsible for drought tolerance in the rice landrace N22.


A phase I and pharmacokinetic study of a powder-filled capsule formulation of oral irinotecan (CPT-11) given daily for 5 days every 3 weeks in patients with advanced solid tumors.
Intravenous (i.v.) irinotecan is a cytotoxic topoisomerase I inhibitor with broad clinical activity in metastatic colorectal cancer and other tumors. The development of an oral formulation of irinotecan could enhance convenience and lessen the expense of palliative irinotecan delivery. This phase I study evaluated the dose-limiting toxicities (DLT), maximum tolerated dose (MTD), and pharmacokinetics (PK) of irinotecan given as a powder-filled capsule (PFC) daily for 5 days every 3 weeks. Patients with advanced solid tumors received escalating doses of oral irinotecan daily for 5 days every 3 weeks. Plasma samples were collected following the first and fifth doses of irinotecan during Cycle 1 to determine the PK of irinotecan and its major circulating metabolites: SN-38, SN-38G, and APC. 20 patients (median age 61.5 years, range 40-75; M/F 12/8; ECOG PS 0=5, 1=11, 2=4) received oral irinotecan at dose levels of 30 (n=3), 40 (n=3), 50 (n=6), and 60 (n=8) mg/m(2)/day. Of the eight patients enrolled at 60 mg/m(2), three patients experienced DLT (> or = grade 3) consisting of nausea (three patients), vomiting (three patients), diarrhea (two patients), and febrile neutropenia (two patients) for which all the three patients required hospitalization. Treatment of six patients at the 50-mg/m(2) dose level resulted in no DLT. Other toxicities observed include abdominal pain, alopecia, anorexia, and asthenia. After oral administration, irinotecan was rapidly absorbed into systemic circulation and converted to the active metabolite SN-38. Increasing dose levels resulted in a dose-dependent increase in mean exposure parameters (Cmax and AUC) of irinotecan and metabolites. Systemic exposure parameters (Cmax and AUC(0-24)) of irinotecan and SN-38 were comparable between days 1 and 5. The extent of conversion from irinotecan to SN-38 was approximately threefold higher after the oral administration compared to that previously observed after i.v. administration. The exposure parameters of irinotecan or SN-38 are of limited value in predicting severity of Cycle 1 toxicities in the twofold dose range evaluated. Daily oral administration of irinotecan as the PFC formulation for 5 days every 3 weeks can safely deliver protracted exposure to SN-38, with the MTD of 50 mg/m(2)/d.


Raccoon dog (Nyctereutes procyonoides)--the new host of Echinococcus multilocularis in Poland.
The alimentary canals of 9 species of domesticated or wild animals were examined for the presence of E. multilocularis. The tapeworm was found in 9 red foxes (Vulpes vulpes) out of 155 examined (5.8%) and in 2 raccoon dogs (Nyctereutes procyonoides) of 25 examined (8.0%). Our studies proved that in Poland the raccoon dog can also be infected with E. multilocularis.


General Anesthesia for Pediatric Radiation Therapy in the Era of COVID-19.
Managing pediatric patients requiring daily general anesthesia (GA) for radiation therapy (RT) in the setting of COVID-19 is complex, owing to the aerosolizing nature of GA procedures, the risk of cardiopulmonary complications for infected patients, and the treatment of immunocompromised oncology patients in a busy, densely populated radiation oncology clinic. We developed an institutional protocol to define procedures for COVID-19 testing and protection of patients, caregivers, and staff, hypothesizing that this protocol would allow patients requiring GA to be safely treated, minimizing COVID-19 transmission risk to both patients and staff, and at the same time maintaining pre-COVID-19 patient volumes. All patients underwent COVID-19 testing before their first treatment and thrice weekly during treatment. For patients who tested positive for COVID-19, RT was delivered in the last end-of-day treatment appointment. A negative pressure room was used for GA induction and recovery, and separate physician/nurse teams were designated for in-room versus out-of-room patient management. Seventy-eight pediatric patients received RT under GA, versus 69 over the same prior year timeframe, and 2 patients received 2 courses of RT under GA, for a total of 80 courses. The mean age was 4.9 years (range, 0.5-19.0 years) and 41 of 78 (52.6%) were male. Two patients (2.6%) received 2 courses of RT under GA, establishing a total of 80 courses. The mean number of treatment fractions was 22.2 (range, 1-40). Two of 78 patients (2.6%) tested positive for COVID-19; both were asymptomatic. Both patients completed treatment as prescribed. Neither patient developed cardiopulmonary symptoms complicating anesthesia, and neither patient experienced grade 3+ acute radiation toxicity. With careful multidisciplinary planning to mitigate COVID-19 risk, pediatric RT with GA was carried out for a large patient volume without widespread infection and without increased toxic effects from either GA or RT.


Car-Parrinello molecular dynamics study of the intramolecular vibrational mode-sensitive double proton-transfer mechanisms in porphycene.
Car-Parrinello molecular dynamics simulations were carried out to help interpret proton-transfer processes observed experimentally in porphycene under thermodynamic equilibrium conditions (NVT ensemble) as well as during selective, nonequilibrium vibrational excitations of the molecular scaffold (NVE ensemble). In the NVT ensemble, the population of the trans form in the gas phase at 300 K is 96.5%, and of the cis-1 form is 3.5%, in agreement with experimental data. Approximately 70% of the proton-transfer events are asynchronous double proton transfers. According to the high resolution simulation data they consist of two single transfer events that rapidly take place one after the other. The average time-period between the two consecutive jumps is 220 fs. The gas phase reaction rate estimate at 300 K is 3.6 ps, which is comparable to experimentally determined rates. The NVE ensemble nonequilibrium ab initio MD simulations, which correspond to selective vibrational excitations of the molecular scaffold generated with high resolution laser spectroscopy techniques, exhibit an enhancing property of the 182 cm(-1) vibrational mode and an inhibiting property of the 114 cm(-1) one. Both of them influence the proton-transfer rate, in qualitative agreement with experimental findings. Our ab initio simulations provide new predictions regarding the influence of double-mode vibrational excitations on proton-transfer processes. They can help in setting up future programmable spectroscopic experiments for the proton-transfer translocations.


[ArticlThe application of glucocorticoid hormones for correction of spermatogenesis шт males with endocrine-dependent forms of infertility. eTitle].
Among main tasks of andrology, besides search of new pharmaceuticals, an important place takes the profound analysis of pathogenesis of formation of infertility in males in every particular case with purpose of selection of pathogenetically justified therapy. The article presents the results of application of glucocorticoid medications for normalization of indices of spermogram in males with reproduction disorders. Two age groups were examined. The first group consisted of 45 individuals of 21-35 years and the second group included 30 individuals of 36-45 years. All males were applied standard clinical laboratory analysis examination including andrologist's examination, ltrasound examination of organs of scrotum and prostate, macro- and microscopic analysis of ejaculate, hormonal analysis. The treatment of patients consisted of application of medication of glucocorticoid hormones - Dexamethasonum with dosage depending on expression of hormonal alterations (0.25 mg - 1.5 mg per day). In males with supra-renal hyper-androgeny application of glucocorticoid hormones demonstrated higher efficiency of hormone therapy for correction of the given condition at absence of any side-effects that permitted to ameliorate reproductive function by normalizing indices of spermatogenesis and level of supra-renal hormones. At the given mode of treatment of pathozoospermia in males with supra-renal hyper-androgeny occurs increasing of concentration of active spermatozoa participating in fertilization and also terms of vitality of spermatozoa and activity of spermatogenesis. It is established that similar treatment shows no side-effects on organism and it has no significant contraindications for application.


Reliability and validity of outcome measures of in-hospital and at-home visits in a randomized, double-blind, placebo-controlled trial for spinal muscular atrophy.
We used at-home assessments in a clinical trial to relieve the visit burden for participants. A total of 57 patients with type II or III spinal muscular atrophy were enrolled and 10 of them (7 type II and 3 type III) received at-home assessments. The primary end points were Gross Motor Function Measure, Manual Muscle Test, and serum biomarker. The secondary endpoints were Modified Hammersmith Functional Motor Scale and forced vital capacity. The correlation coefficients and analysis of covariance showed good reliability and validity of all outcome measures. Except for Gross Motor Function Measure and Modified Hammersmith Functional Motor Scale, there were no significant differences in measures between in-hospital and at-home groups (intersubject) or among 3 patients who received both at-home and in-hospital visits (intrasubject). We concluded that at-home assessments could provide sufficient reliability in a controlled trial. This modification could help design a successful clinical trial for spinal muscular atrophy.


The PCBP1 gene encoding poly(rC) binding protein I is recurrently mutated in Burkitt lymphoma.
The genetic hallmark of Burkitt lymphoma is the translocation t(8;14)(q24;q32), or one of its light chain variants, resulting in IG-MYC juxtaposition. However, these translocations alone are insufficient to drive lymphomagenesis, which requires additional genetic changes for malignant transformation. Recent studies of Burkitt lymphoma using next generation sequencing approaches have identified various recurrently mutated genes including ID3, TCF3, CCND3, and TP53. Here, by using similar approaches, we show that PCBP1 is a recurrently mutated gene in Burkitt lymphoma. By whole-genome sequencing, we identified somatic mutations in PCBP1 in 3/17 (18%) Burkitt lymphomas. We confirmed the recurrence of PCBP1 mutations by Sanger sequencing in an independent validation cohort, finding mutations in 3/28 (11%) Burkitt lymphomas and in 6/16 (38%) Burkitt lymphoma cell lines. PCBP1 is an intron-less gene encoding the 356 amino acid poly(rC) binding protein 1, which contains three K-Homology (KH) domains and two nuclear localization signals. The mutations predominantly (10/12, 83%) affect the KH III domain, either by complete domain loss or amino acid changes. Thus, these changes are predicted to alter the various functions of PCBP1, including nuclear trafficking and pre-mRNA splicing. Remarkably, all six primary Burkitt lymphomas with a PCBP1 mutation expressed MUM1/IRF4, which is otherwise detected in around 20-40% of Burkitt lymphomas. We conclude that PCBP1 mutations are recurrent in Burkitt lymphomas and might contribute, in cooperation with other mutations, to its pathogenesis.


Three municipal solid waste gasification technologies analysis for electrical energy generation in Brazil.
In Brazil, in 2016, 196,050 tonnes day-1 of municipal solid waste (MSW) were collected, which means a waste generation of 1.035 kg per capita per day. Only 59.1% of the waste has adequate destination in sanitary landfills, whereas the remaining 40.9% has inadequate destination in controlled landfills and open dumps (ABRELPE, 2018). Among all the states in the country, the State of São Paulo has the biggest per capita generation: 2.290 kg. Today, the only waste destination practiced in the country is deposition in landfills, but other possibilities can be considered. Among thermal treatment routes, the gasification of MSW is an interesting alternative to be studied, because of its versatility and relatively low emissions. The aim of this work is to evaluate the potential of electricity generation through MSW gasification in Santo André city, Brazil, comparing three waste gasification technologies: TPS Termiska Processer AB, Carbogas and Energos. These alternatives have operated commercially for a few years, and data are available. Specific characteristics of each technology were taken into account, such as the reactor type and fuel properties. For the electricity production scheme, two energy conversion systems were assumed: an internal combustion engine and a steam power cycle. From the process parameters adopted, the results showed that Carbogas technology, coupled to internal combustion engines, presents the highest efficiency of electricity generation (30%) and also the lowest cost of electrical energy produced (US$65.22 MWh-1) when Santo André's gate fee is applied.


Practical anatomic landmarks for determining the insertion depth of central venous catheter in paediatric patients.
Various methods have been recommended to decide a proper insertion depth of central venous catheter (CVC). The carina is recommended as a useful target level for the CVC tip position. We evaluated the sternal head of a right clavicle and the nipples as anatomic landmarks for determining the optimal depth of CVC in paediatric patients. Ninety children, <5 yr, undergoing catheterization through the right internal jugular vein were enrolled. The insertion depth was determined as follows. The insertion point was designated as 'Point I'. The sternal head of the right clavicle was called 'Point A' and the midpoint of the perpendicular line drawn from Point A to the line connecting both nipples was called 'Point B'. The insertion depth of CVC was determined by adding the two distances (from I to A and from A to B) and subtracting 0.5 cm from this. A chest radiography was taken and the distance of the CVC tip from the carina level was measured by the Picture Archiving and Communicating System. The mean distance of the CVC tip from the carina level was 0.1 (1.0) (P=0.293) cm above the carina (95% CI 0.1 cm below the carina-0.3 cm above the carina). There was no specific relationship between the distance of the CVC tip from the carina level and the patients' age, height, and weight. The CVC tip could be placed near the carina by using the external landmarks without any formulae, images, and devices in children in our study.


Independent verification of monitor unit calculation for radiation treatment planning system.
To ensure the accuracy of dose calculation for radiation treatment plans is an important part of quality assurance (QA) procedures for radiotherapy. This study evaluated the Monitor Units (MU) calculation accuracy of a third-party QA software and a 3-dimensional treatment planning system (3D TPS), to investigate the feasibility and reliability of independent verification for radiation treatment planning. Test plans in a homogenous phantom were designed with 3-D TPS, according to the International Atomic Energy Agency (IAEA) Technical Report No. 430, including open, blocked, wedge, and multileaf collimator (MLC) fields. Test plans were delivered and measured in the phantom. The delivered doses were input to the QA software and the independent calculated MUs were compared with delivery. All test plans were verified with independent calculation and phantom measurements separately, and the differences of the two kinds of verification were then compared. The deviation of the independent calculation to the measurements was (0.1 +/- 0.9)%, the biggest difference fell onto the plans that used block and wedge fields (2.0%). The mean MU difference between the TPS and the QA software was (0.6 +/- 1.0)%, ranging from -0.8% to 2.8%. The deviation in dose of the TPS calculation compared to the measurements was (-0.2 +/- 1.7)%, ranging from -3.9% to 2.9%. MU accuracy of the third-party QA software is clinically acceptable. Similar results were achieved with the independent calculations and the phantom measurements for all test plans. The tested independent calculation software can be used as an efficient tool for TPS plan verification.


Stability of mixed micellar systems made by solubilizing phosphatidylcholine-cholesterol vesicles by bile salts.
Complete solubilization of phosphatidylcholine and cholesterol by bile salts in the form of stable mixed micelles requires that the effective ratio of bile salt/lipids in the mixed micelles (Re = ([bile salt] - critical micellar concentration)/([phosphatidylcholine] + [cholesterol]) will exceed a critical value. This equilibrium solubilizing ratio is an increasing function of the cholesterol/phosphatidylcholine ratio. In contrast, the concentration of sodium cholate required for solubilization of vesicles made of phosphatidylcholine and cholesterol does not increase by increasing the cholesterol/phosphatidylcholine ratio. Consequently, the latter solubilization procedure yields metastable mixed micelles whenever the cholate concentration is higher than that required for vesicle solubilization but lower than that needed for establishing a micellar equilibrium. These metastable mixed micelles undergo partial revesiculation to form cholesterol-rich vesicles that subsequently aggregate. Cholesterol crystallization appears to occur through its reorganization within these aggregated vesicles. The overall rate of the above series of processes increases sharply with the total lipid concentration and with the cholesterol/phosphatidylcholine ratio. The dependence of the rate on the effective ratio of bile salts/lipids is very complex: at any given ratio of cholesterol/phosphatidylcholine within the range of 0.3 to 0.5, increasing the cholesterol/phosphatidylcholine ratio requires higher cholate concentrations for the formation of stable mixed micelles (higher equilibrium solubilizing ratio). On the other hand, the metastable mixed micellar larsystems are long-lived whenever the effective ratio of cholate/lipids is lower than a critical value.(ABSTRACT TRUNCATED AT 250 WORDS)


Enzyme-linked immunosorbent assays for measurement of reovirus immunoglobulin G, A, and M levels in serum.
Enzyme-linked immunosorbent assays were developed for the quantitation of anti-reovirus immunoglobulin G (IgG), IgA, and IgM in serum. The assays were specific, sensitive, and reproducible and measured antibodies to the reovirus group antigen. They should permit diagnosis of recent reovirus infection and longitudinal study of the development of reovirus group antibodies.


The Depressed Frail Phenotype: The Clinical Manifestation of Increased Biological Aging.
Depression in later life is a severe public health problem, associated with higher rates of mortality, suicide, and dementia. Effectiveness of treatment is limited by the failure to deconstruct the heterogeneity of the illness and because diagnostic criteria, pathophysiological models, and treatment algorithms for depression are primarily based on studies of younger adults even though symptoms of the illness and physiology of the patient change with age. Thus, understanding how aging interacts with depressive illness may elucidate endophenotypes of late-life depression with different clinical manifestations and underlying mechanisms that can then be targeted with more personalized approaches to treatment. This paper proposes a model for the critical confluence between depression and frailty, a high-risk morbidity and mortality syndrome of later life. This model hypothesizes that characteristics of frailty in adults with late life depression represent the clinical manifestation of greater biological aging and their presence in the context of a depressive illness exposes elders to deleterious trajectories. Potential common biological substrates that may result in the manifestation of the depressed frail phenotype including mitochondrial functioning, dopaminergic neurotransmission, and inflammatory processes and implications for the assessment and treatment of adults with late-life depression are discussed. As society continues to live longer, the preservation of the quality of these added years becomes paramount, and the combined impact of depression and frailty on the preservation of this quality warrants the attention of clinical researchers and physicians.


Systemic sclerosis-associated pulmonary hypertension: short- and long-term effects of epoprostenol (prostacyclin).
To evaluate the short- and long-term effects of intravenous epoprostenol in patients with pulmonary hypertension (PH) associated with systemic sclerosis (SSc). Sixteen patients with SSc-associated PH and New York Heart Association (NYHA) class III or IV symptomatology underwent right heart catheterization for determination of baseline hemodynamic values. Vasoreactivity was assessed with either inhaled nitric oxide or intravenous adenosine. After a medication washout period, all patients received intravenous epoprostenol in incrementally increasing doses; tolerance was assessed according to symptoms and hemodynamic findings at each dose increment and at the conclusion of the medication trial. Once a stable medication regimen was established, patients were discharged and followed up as outpatients for assessment of symptoms and exercise tolerance as measured by change in the NYHA class. Repeat hemodynamic testing was performed in 4 patients at 1 year and in 2 patients at 2 years of treatment. Therapeutic response to epoprostenol, defined by a reduction in the pulmonary vascular resistance of > or =25%, was achieved in the short-term treatment period in 13 of 16 patients (81.3%). Improvement in symptoms and exercise tolerance occurred in all patients, and a significant short-term hemodynamic response was observed. Followup hemodynamic tests revealed persistent favorable responses in all 4 of the patients studied. Most patients with PH secondary to SSc manifest favorable hemodynamic responses to epoprostenol in the short term. Long-term epoprostenol was generally well tolerated and provides a potential therapeutic option for patients with PH secondary to SSc.


Engineering the public-use reinforced concrete buildings of Ankara during the Early Republic of Turkey, 1923-1938.
Today, reinforced concrete (RC) is the most commonly used construction material in Turkey. It first emerged in Europe in the 1850s and was adopted in a number of Late Ottoman period structures, mostly in İstanbul, during the first two decades of the twentieth century. During the Early Turkish Republic (1923-1938), RC appeared in public-use buildings in Ankara, such as the Ethnographic Museum, which was the first in the new capital to feature RC elements, leading the way for many more structures to come. Despite the fact that Turkish and foreign civil engineers faced a series of economic, social, cultural, political, educational and technical challenges during the transition from masonry and timber construction to RC, its adoption was facilitated by the fact that as a European building technology, it became symbolically important to the new republic. Equated with modernity, RC would allow its capital, Ankara, to construct an identity that would contrast with İstanbul, the capital of the Ottoman Empire. This transition would also be catalyzed by the rise of a professional class of Turkish civil engineers who deployed RC to reinforce their authority as trained specialists and agents of modernization.


Substrate binding causes movement in the ATP binding domain of Escherichia coli adenylate kinase.
Crystallographic evidence suggests that there is a large hinged domain motion associated with substrate binding in adenylate kinase. To test this hypothesis, resonance energy transfer measurements of substrate binding were initiated. Adenylate kinase from Escherichia coli consists of three domains: the main body of the enzyme with alpha-helical and beta-sheet secondary structure, and domains that close over the AMP and ATP binding sites. Four single tryptophan mutants were constructed to map distances. Two tryptophan mutants were positioned at residues 133 (Y133W) and 137 (F137W), which are in the domain that closes over the ATP binding site. Mutant F86W that is located at the AMP binding site, and mutant S41W that is in the loop that close over AMP, complete the mapping library. Energy transfer was measured between each of these tryptophans and 5-[[2-(acetylamino)ethyl]amino]naphthalene-1-sulfonic acid (AEDANS) covalently bound to the single cysteine residue at position 77, which is located in the main body of adenylate kinase. The distance between the tryptophan of the F137W mutant adenylate kinase and the AEDANS-labeled Cys-77 decreased by 12.1 A upon the binding of the bisubstrate inhibitor P1, P5-bis(5'-adenosyl) pentaphosphate (AP5A). There were only small alterations in the tryptophan to Cys-77-AEDANS distances in the Y133W, F86W, and S41W mutants upon the binding of AP5A, ATP, or AMP, implying that movement of residues 133, 86, and 41 in relation to the Cys-77 residue was minimal. These results suggest that there is significant closure of the ATP binding domain upon the binding of ATP or AP5A. Unexpectedly, exposure of the enzyme to AMP also introduced a partial closure of the ATP hinged domain.


Patient specific anatomy: the new area of anatomy based on computer science illustrated on liver.
Over the past century, medical imaging has brought a new revolution: internal anatomy of a patient could be seen without any invasive technique. This revolution has highlighted the two main limits of current anatomy: the anatomical description is physician dependent, and the average anatomy is more and more frequently insufficient to describe anatomical variations. These drawbacks can sometimes be so important that they create mistakes but they can be overcome through the use of 3D patient-specific surgical anatomy. In this article, we propose to illustrate such improvement of standard anatomy on liver. We first propose a general scheme allowing to easily compare the four main liver anatomical descriptions by Takasaki, Goldsmith and Woodburne, Bismuth and Couinaud. From this general scheme we propose four rules to apply in order to correct these initial anatomical definitions. Application of these rules allows to correct usual vascular topological mistakes of standard anatomy. We finally validate such correction on a database of 20 clinical cases compared to the 111 clinical cases of a Couinaud article. Out of the 20 images of the database, we note a revealing difference in 14 cases (70%) on at least one important branch of the portal network. Only six cases (30%) do not present a revealing difference between both labellings. We also show that the right portal fissure location on our 20 cases defined between segment V and VI of our anatomical definition is well correlated with the real position described by Couinaud on 111 cases, knowing that the theoretical position was only found in 46 cases out of 111, i.e., 41.44% of cases with the non-corrected Couinaud definition. We have proposed a new anatomical segmentation of the liver based on four main rules to apply in order to correct topological errors of the four main standard segmentations. Our validation clearly illustrates that this new definition corrects the large amount of mistakes created by the current standard definitions, increased by physician interpretation that can vary from one case to another.


Influence of Al₂O₃ Nanoparticle Addition on a UV Cured Polyacrylate for 3D Inkjet Printing.
The brittleness of acrylic photopolymers, frequently used in 3D Inkjet printing, limits their utilization in structural applications. In this study, a process was developed for the production and characterization of an alumina-enhanced nanocomposite with improved mechanical properties for Inkjet printing. Ceramic nanoparticles with an average primary particle size (APPS) of 16 nm and 31 nm, which was assessed via high-resolution scanning electron microscopy (HRSEM), were functionalized with 3.43 and 5.59 mg/m² 3-(trimethoxysilyl)propyl methacrylate (MPS), respectively, while being ground in a ball mill. The suspensions of the modified fillers in a newly formulated acrylic mixture showed viscosities of 14 and 7 mPa∙s at the printing temperature of 60 °C. Ink-jetting tests were conducted successfully without clogging the printing nozzles. Tensile tests of casted specimens showed an improvement of the tensile strength and elongation at break in composites filled with 31 nm by 10.7% and 74.9%, respectively, relative to the unfilled polymer.


Hormones and Pod Development in Oilseed Rape (Brassica napus).
The endogenous levels of several plant growth substances (indole acetic acid, IAA; abscisic acid, ABA; zeatin, Z; zeatin riboside, [9R]Z; isopentenyladenine, iP; and isopentenyladenosine, [9R]iP were measured during pod development of field grown oilseed Rape (Brassica napus L. var oleifera cv Bienvenu) with high performance liquid chromatography and immunoenzymic (enzyme-linked immunosorbent assay, ELISA) techniques. Results show that pod development is characterized by high levels of Z and [9R]Z in 3 day old fruits and of IAA on the fourth day. During pod maturation, initially a significant increase of IAA and cytokinins was observed, followed by a progressive rise of ABA levels and a concomitant decline of IAA and cytokinin (except iP) levels. The relationship between hormone levels and development, especially pod number, seed number per pod, and seed weight determination, will be discussed.


Health-related characteristics of men who have sex with men: a comparison of those living in "gay ghettos" with those living elsewhere.
This study investigated the limitations of probability samples of men who have sex with men (MSM), limited to single cities and to the areas of highest concentrations of MSM ("gay ghettos"). A probability sample of 2881 MSM in 4 American cities completed interviews by telephone. MSM who resided in ghettos differed from other MSM, although in different ways in each city. Non-ghetto-dwelling MSM were less involved in the gay and lesbian community. They were also less likely to have only male sexual partners, to identify as gay, and to have been tested for HIV. These differences between MSM who live in gay ghettos and those who live elsewhere have clear implications for HIV prevention efforts and health care planning.


Increased throughput for low-abundance protein biomarker verification by liquid chromatography/tandem mass spectrometry.
Low-abundance protein quantification has historically been performed using ligand binding techniques. However, due to the time and cost associated with developing enzyme-linked immunosorbent assay (ELISA), mass spectrometric approaches are playing an increasingly important role. Protein quantification at or below the nanogram per milliliter level using liquid chromatography/tandem mass spectrometry (LC/MS/MS) typically utilizes an immunoaffinity (IA) enrichment step such as immunoprecipitation. In order to maximize mass spectrometry (MS) sensitivity, protein enrichment is followed by a proteolytic cleavage step used to generate a surrogate peptide with better mass spectrometric properties. Unlike ELISA, IA-LC/MS/MS is a serial technique that can require up to 3 days for a single batch analysis due to lengthy incubation and digestion steps. This report describes the use of immunoprecipitation in 96-well ELISA format (IPE) and microwave-assisted protein digestion to reduce the time required to perform LC/MS/MS protein analyses to within a single day. The utility of this approach was investigated through its application to previously published LC/MS/MS protein assays from our laboratory for two cardiotoxicity biomarkers, Myl3 and NTproBNP. Using commercially available antibodies, IPE and microwave-assisted digestion were used to repeat intraday validations for these markers, and intraday precision (%CV) and accuracy (%RE) did not exceed 11% or 3% for either assay, respectively. Additionally, lower limits of quantification of 100 pg/mL (NTproBNP) and 0.95 ng/mL (Myl3) were achieved.


Total Joint Arthroplasty in the Morbidly Obese: How Body Mass Index ≥40 Influences Patient Retention, Treatment Decisions, and Treatment Outcomes.
The United States is in an obesity epidemic. Obesity has multiple common comorbid conditions, including lower extremity arthritis. We sought to examine the course of treatment for a population with body mass index (BMI) ≥40 kg/m2 and osteoarthritis (OA) of the hip or knee. We investigated decision criteria that influenced arthroplasty surgeons to recommend nonoperative management vs total joint arthroplasty (TJA). For those patients who ultimately received TJA, we compared outcomes in this population to those with BMI <40 kg/m2. This study retrospectively reviewed 158 new patients with BMI ≥40 kg/m2 and moderate/severe OA of the hip or knee. Demographics, comorbidity profiles, and weight loss were compared between groups that underwent TJA and those that did not. The arthroplasty database was used to identify patients who underwent TJA during 2016-2018 (N = 1473). Comorbidities, readmissions, surgical site infections, and overall complications were compared between those with BMI ≥40 kg/m2 and BMI <40 kg/m2. About 51.3% of new patients with BMI ≥40 kg/m2 and moderate/severe OA did not return for a second clinic visit. Of those who did return, 42.9% eventually underwent surgery. BMI was higher in single visit patients vs those with multiple visits (49.5 vs 46.3 kg/m2, P < .001), no difference in those scheduled on an "as-needed" basis vs a specific return date (P = .18), and did not change significantly during the 2-year follow-up (P = .41). Patients who underwent TJA had a lower mean BMI at presentation than their nonoperative counterparts (44.5 vs 47.6 kg/m2, P < .01) and demonstrated significant weight loss prior to surgery (44.5 vs 42.6 kg/m2, P < .05). When comparing patients with BMI ≥40 kg/m2 vs BMI <40 kg/m2, overall complications were not higher in the BMI ≥40 kg/m2 group, although surgical site infections were higher in those undergoing total hip arthroplasty with BMI ≥40 kg/m2 (0.3% vs 3.1%, P < .05). A majority of patients with BMI ≥40 kg/m2 and moderate/advanced OA will be lost to orthopedic follow-up. A relatively lower BMI indicates a greater chance of retention in care, and ultimately surgery, but does not influence surgeons' recommendations to continue orthopedic management. Patients who persist in seeking treatment, lose significant weight, and exhaust nonoperative alternatives may be suitable for TJA despite a BMI ≥40 kg/m2, with an overall complication rate of 4.3%. However, only 9% of patients at 2-year follow-up achieved BMI <40 kg/m2 and only 20% of surgeries were performed on patients who had achieved this proposed cutoff.


Addressing Interpersonal Violence as a Health Policy Question Using Interprofessional Community Educators.
The health effects of intimate partner violence (IPV) not only include physical injury, but can also manifest as posttraumatic stress disorder, anxiety, and others. US medical students report receiving inadequate training about IPV. This case-based tutorial for third-year medical students examines: (1) a clinical encounter with a patient experiencing several complex challenges including IPV and homelessness; (2) the implications of existing policy on the delivery of health care services; and (3) the impact of policies on patient choices. This case is completed during a family medicine clerkship. The 2-hour case review moves between small- and large-group sessions led by community interprofessional experts at a local family advocacy center. Optimal group size is three to four students and one or two experts per group. The large-group session should be led by a dynamic moderator who is familiar with the Socratic method of teaching to elicit a variety of responses to ad hoc challenge questions. Materials provided include student resources, student case, facilitator guide, moderator guide, and sample brochure of IPV documentation policies. To date, over 200 students have participated in this session. During the most recent iteration the average response to the question, "As a result of the FAC experience, I feel more empowered to care for persons experiencing IPV," was 4.1 out of 5 (5 = strongly agree). Public health, health policy, and clinical topics can be effectively taught by an interprofessional team of community experts and lead to improved student understanding of the importance of health policy to both individual and population health outcomes.


The neurofibromatosis type I gene promotes autophagy via mTORC1 signalling pathway to enhance new bone formation after fracture.
Bone fracture is one of the most common injuries. Despite the high regenerative capacity of bones, failure of healing still occurs to near 10% of the patients. Herein, we aim to investigate the modulatory role of neurofibromatosis type I gene (NF1) to osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) and new bone formation after fracture in a rat model. We studied the NF1 gene expression in normal and non-union bone fracture models. Then, we evaluated how NF1 overexpression modulated osteogenic differentiation of BMSCs, autophagy activity, mTORC1 signalling and osteoclastic bone resorption by qRT-PCR, Western blot and immunostaining assays. Finally, we injected lentivirus-NF1 (Lv-NF1) to rat non-union bone fracture model and analysed the bone formation process. The NF1 gene expression was significantly down-regulated in non-union bone fracture group, indicating NF1 is critical in bone healing process. In the NF1 overexpressing BMSCs, autophagy activity and osteogenic differentiation were significantly enhanced. Meanwhile, the NF1 overexpression inhibited mTORC1 signalling and osteoclastic bone resorption. In rat non-union bone fracture model, the NF1 overexpression significantly promoted bone formation during fracture healing. In summary, we proved the NF1 gene is critical in non-union bone healing, and NF1 overexpression promoted new bone formation after fracture by enhancing autophagy and inhibiting mTORC1 signalling. Our results may provide a novel therapeutic clue of promoting bone fracture healing.


Hypothalamic-pituitary-thyroid (HPT) axis functioning in anxiety disorders. A systematic review.
Depression has repeatedly been linked to subclinical hypothyroidism, and thyroid hormones have successfully been used to augment antidepressant treatment. By contrast, the extent of thyroid dysfunction in anxiety disorders remains less clear. This is surprising, given that anxiety-related symptoms (e.g., nervousness, palpitations, increased perspiration) are highly prevalent in hyperthyroidism. The present study was undertaken to synthesize the literature on hypothalamic-pituitary-thyroid (HPT) axis functioning in anxiety disorders. The PubMed and PsycINFO databases were systematically searched. Three types of studies were included: (1) "comorbidity studies" assessing the prevalence of thyroid disorders in individuals with anxiety disorders, (2) "case-control studies" comparing HPT parameters between patients and controls, and (3) "correlational studies" assessing self-reported anxiety levels and HPT parameters. Risk of bias was assessed via a standardized quality rating. Twenty studies were eligible. Nearly all found the comorbidity between anxiety and thyroid disorders was significant. Half of the studies additionally supported the notion of subtle thyroid dysfunction in that thyroid-stimulating hormone (TSH) responses to the administration of thyrotropin-releasing hormone (TRH) were blunted and an inverse relationship was observed between self-reported anxiety levels and TSH. Overall, HPT assessments were well conducted, but several studies failed to adjust their analyses for smoking, body mass index (BMI), and depression. The findings resonate well with clinical recommendations to routinely screen for thyroid disorders in patients with anxiety disorders, and with what is known from basic research about thyroid-brain interactions. The results of the risk of bias assessment underscore the importance of further high-quality experimental and longitudinal epidemiological research.


Adaptive changes in enzyme activity and metabolic pathways in adipose tissue from meal-fed rats.
A number of metabolic factors and the activity of a number of enzymes were determined in meal-fed (animals fed a single daily 2 hr meal) and nibbling (ad libitum-fed) rats. The dependency of the observed adaptive changes on the ingestion of carbohydrate was studied by feeding diets high in carbohydrate or fat. Glucose-6-phosphate dehydrogenase and NADP-malic dehydrogenase were more active in adipose tissue from high carbohydrate meal-fed rats than in tissue from ad libitum-fed rats. The activity in adipose tissue of isocitric dehydrogenase, 6-phosphogluconate dehydrogenase, and NAD-malic dehydrogenase did not increase significantly in response to meal-feeding the high carbohydrate diet. No increase in lipogenesis or enzyme activity could be demonstrated in adipose tissue from rats meal-fed a high fat diet. Lipase activity of adipose tissue was increased by high carbohydrate meal-feeding and decreased by feeding a high fat diet. The in vitro uptake of palmitate-1-(14)C by adipose tissue was depressed by a high fat diet and enhanced in rats meal-fed a high carbohydrate diet. Diaphragm or slices of liver from high fat-fed rats oxidized palmitate-1-(14)C more rapidly than did tissue from ad libitum-fed animals. Evidence is presented for the quantitative importance of citrate as a source of extramitochondrial acetyl CoA in adipose tissue of meal-eating and ad libitum-fed rats. The relationship of extramitochondrially formed citrate to the NAD-malic dehydrogenase-malic enzyme system in adipose tissue is discussed.


Surgical and anatomic factors predict development of leptomeningeal disease in patients with melanoma brain metastases.
Leptomeningeal disease (LMD) is a devastating complication of systemic malignancy, of which there is an unclear etiology. The aim of this study is to determine if surgical or anatomic factors can predict LMD in patients with metastatic melanoma. A retrospective chart review was performed of 1162 patients treated at single institution for melanoma brain metastases (MBM). Patients with fewer than 3 months follow-up or lacking appropriate imaging were excluded. Demographic information, surgical, and anatomic data were collected. Eight hundred and twenty-seven patients were included in the final review. On multivariate analysis for the entire cohort, female gender, dural-based and intraventricular metastasis, and tumor bordering CSF spaces were associated with increased risk of LMD. Surgical resection was not significant for risk of LMD. On multivariate analysis of patients who have undergone surgical resection of a metastatic tumor, dural-based and intraventricular metastasis, ventricular entry during surgery, and metastasis in the infratentorial space were associated with increased risk of LMD. On multivariate analysis of patients who did not undergo surgery, chemotherapy after initial diagnosis and metastasis bordering CSF spaces were associated with increased risk of LMD. In a single-institution cohort of MBM, we found that surgical resection alone did not result in an increased risk of LMD. Anatomical factors such as dural-based and intraventricular metastasis were significant for developing LMD, as well as entry into a CSF space during surgical resection. These data suggest a strong correlation between anatomic location and tumor cell seeding in relation to the development of LMD.


Necrotizing myositis: highlighting the hidden depths - case series and review of the literature.
Necrotizing myositis (NM) is a life-threatening emergency. It causes focal muscle necrosis without abscess formation or extensive involvement of the overlying fascia and soft tissue. It is a clinical diagnosis requiring a high index of clinical suspicion. Usual presentation can readily be mistaken to represent more benign pathologies such as muscular injury, viral myopathy or deep venous thrombosis. The clinical course following initial misdiagnosis is rapid deterioration into profound sepsis and progressive multiorgan failure. Prompt treatment is associated with favourable outcomes, but early diagnosis is challenging due to initial absence of cutaneous signs and symptoms. Delayed referral to surgeons with appropriate expertise results in higher morbidity and mortality. The cornerstones to treatment are complete surgical debridement, intensive care management and accurate antimicrobial therapy. We report four cases of NM demonstrating classical scenarios of initial misdiagnosis, delays in referral and review by an experienced surgeon. A review of the current literature to aid with overall management is also included. Review of literature that revealed the most common presentation was antecedent prodromal flu-like symptoms followed by rapidly progressing focal muscle pain. Patients were initially misdiagnosed followed by rapid deterioration into profound sepsis before surgical opinion was obtained. NM is a rare and potentially fatal disease that must be considered in the differential diagnoses of the young, healthy patient with acute limb pain and fever. A high index of suspicion will facilitate earlier management and reduce morbidity and mortality.


Automated Detection of Periprosthetic Joint Infections and Data Elements Using Natural Language Processing.
Periprosthetic joint infection (PJI) data elements are contained in both structured and unstructured documents in electronic health records and require manual data collection. The goal of this study is to develop a natural language processing (NLP) algorithm to replicate manual chart review for PJI data elements. PJI was identified among all total joint arthroplasty (TJA) procedures performed at a single academic institution between 2000 and 2017. Data elements that comprise the Musculoskeletal Infection Society (MSIS) criteria were manually extracted and used as the gold standard for validation. A training sample of 1208 TJA surgeries (170 PJI cases) was randomly selected to develop the prototype NLP algorithms and an additional 1179 surgeries (150 PJI cases) were randomly selected as the test sample. The algorithms were applied to all consultation notes, operative notes, pathology reports, and microbiology reports to predict the correct status of PJI based on MSIS criteria. The algorithm, which identified patients with PJI based on MSIS criteria, achieved an f1-score (harmonic mean of precision and recall) of 0.911. Algorithm performance in extracting the presence of sinus tract, purulence, pathologic documentation of inflammation, and growth of cultured organisms from the involved TJA achieved f1-scores that ranged from 0.771 to 0.982, sensitivity that ranged from 0.730 to 1.000, and specificity that ranged from 0.947 to 1.000. NLP-enabled algorithms have the potential to automate data collection for PJI diagnostic elements, which could directly improve patient care and augment cohort surveillance and research efforts. Further validation is needed in other hospital settings. Level III, Diagnostic.


Effects of Dexamethasone on DNA Synthesis in Lens Epithelial Cells Are Dependent on Cell Type and Growth Factor.
To determine the factors that influence the ability of dexamethasone (dex) to inhibit or stimulate the growth of lens epithelial cells. Different growth factors with or without dex (10-6 M) were added to quiescent cultures of two clones of Nakano mouse lens epithelial cells (NK11) in serum-free medium. DNA synthesis was then measured after 8-12 hours by the incorporation of tritiated thymidine. Dex was found to both stimulate and inhibit mitogen-induced 3H-thymidine incorporation into the DNA of cultured mouse lens epithelial cells. Enhancement or repression by dex was found to depend on the growth factor used to stimulate the quiescent cell. EGF and insulin were consistently inhibited with dex. Basic fibroblast growth factor (bFGF) and retinoblastoma-derived growth factor (RbDGF) were both enhanced and inhibited by dex, depending on the growth factor concentration and the cell clone used for the experiment. Additionally, RbDGF protects against the dex inhibition of insulin stimulation, but not the inhibition of EGF stimulation. Progesterone, an inhibitor of the activation of the glucocorticoid receptor, blocks the dex inhibitory effect on the EGF and insulin stimulation of DNA synthesis. The ability of progesterone to affect the dex inhibition is consistent with the dex receptor modulating DNA synthesis. The dex effect on DNA synthesis, either stimulatory or inhibitory, was still seen if dex was added as late as 10 hours after the growth factor. The study demonstrated that dex reduces the overall growth and activity of lens epithelial cells in vitro. This result provides insight into the risk of developing posterior subcapsular cataracts (PSC) in patients on oral glucocorticoid therapy. Understanding the basic mechanisms by which steroids mediate lens cell growth may provide the ability to more accurately predict who will develop PSC. The present studies show the difference in the effect of dex from lens cell to lens cell, but, more importantly, suggest a pattern of dependent variables that might prove useful in such predictions.


Direct thrombin inhibitors.
Direct thrombin inhibitors may offer advantages over indirect thrombin inhibitors in the management of patients with acute coronary syndromes (ACS). Two direct thrombin inhibitors, hirudin and bivalirudin, have been investigated in Phase II and Phase III clinical trials. Based on the results of a meta-analysis of study data from 25,000 patients, hirudin appears to be more effective than unfractionated heparin (UFH) in the treatment of patients with ACS, but it is associated with an increased rate of major bleeding. A meta-analysis of a smaller patient population has suggested that bivalirudin, too, may be more efficacious than UFH and may also be safer.


An Experimental Thermally Deposited Coating for Improved Bonding to Glass-fiber Posts.
To determine whether an experimental thermally deposited siloxane-methacrylate coating for use in industrial scale applications would improve the bond strength of resin-based materials to glass fiber posts (GFPs) without affecting their mechanical properties. An experimental 5% (w/v) solution of methacryloxypropyltrimethoxysilane was prepared. Two types of GFPs (Exacto, Angelus; White Post DC, FGM) were divided into the following groups: S: silane; SA: silane and adhesive; HS: 35% H₂O₂ and silane; HSA: 35% H₂O₂, silane and adhesive; Exp: siloxane-methacrylate coating (Si-O) via post immersion in experimental solution followed by heating; Exp-S: silane after Si-O treatment; Exp-A: adhesive after Si-O treatment; and Exp-SA: silane and adhesive after Si-O treatment. The posts were positioned in a mold to allow insertion of a dual-curing resin core, serially sectioned into beams, and subjected to microtensile bond strength (μTSB) testing. The three-point bending test and SEM/EDX analysis were used to assess the mechanical and surface properties of untreated GFPs that were etched with H₂O₂ or treated with Si-O. Surface treatments affected the μTSB only for the Exacto GFPs. The highest μTBS (MPa) was observed in Exp-S and Exp-SA groups, whereas H₂O₂ etching resulted in intermediate values. The mechanical properties were not affected by surface treatments. Exacto GFPs had significantly higher flexural strength (σf) and flexural modulus (Ef) than did the White Post DC GFPs, but the latter were significantly stiffer (S) than Exacto, regardless of the surface treatment tested. H₂O₂ promoted morphological changes in post surfaces. The experimental treatment promoted deposition of Si onto the post surface, improving bond strengths of Exacto posts. The proposed novel coating technique is a viable procedure for fiber post manufacturers to improve the μTSB of resin-based materials.


Analysis of 76 kb of the chlorella virus PBCV-1 330-kb genome: map positions 182 to 258.
Analysis of 76 kb of newly sequenced DNA, located between map positions 182 and 258 kb in the 330-kb chlorella virus PBCV-1 genome, revealed 175 open reading frames (ORFs) of 65 codons or longer. One hundred and five of these 175 ORFs were considered major ORFs. Twenty-one of the 105 major ORFs resembled proteins in databases including ribonucleotide reductase small subunit, RNase III, thioredoxin, glutaredoxin, protein disulfide isomerase, deoxynucleoside kinase, frog virus 3 ATPase, Acetobacter cellulose synthase, a bacteriophage encoded endonuclease, and two C-5 cytosine DNA methyltransferases. One of the ORFs was the PBCV-1 major capsid protein. The 105 major ORFs were evenly distributed along the genome. One set of ORFs was separated by 543 nucleotides whereas 75 of the ORFs were separated by fewer than 100 nucleotides. Nineteen of the 175 ORFs resembled other PBCV-1 ORFs, suggesting that they represent either gene duplications or gene families.


Seven new species and one new species record of Sabatieria (Nematoda: Comesomatidae) from the continental slope of New Zealand.
Sabatieria is the most abundant nematode genus on the upper continental slope of New Zealand but no Sabatieria species have yet been recorded/described from the region's deep-sea habitats. The present study describes seven new and one known species of this genus from the continental margin of New Zealand. S. bitumen Botelho et al. 2007 was originally described from the South Atlantic Ocean, and is recorded from the first time in the Pacific Ocean. Sabatieria challengerensis sp. n. is characterised by a combination of short cephalic setae, large fovea amphidialis with 3 turns, short arcuate spicules without distal hook, and short tail with cylindrical proximal portion and blunt, rounded distal portion. Sabatieria parapraedatrix sp. n. is characterised by having a slender body, cuticle with striations resulting from fusion of every second lateral row of dots,fovea amphidialis with 2.0-2.5 turns, spicules with distal hook, and short conico-cylindrical tail. Sabatieria bubulba sp. n. is characterised by the presence of a large muscular pharyngeal bulb, secretory-excretory system with large ventral gland at level of intestine and with cuticularised pore opening. Sabatieria exculta sp. n. is characterised by having a slender body, cuticle with faint annulations resulting from fusion of every second or third lateral row of dots, short cephalic setae, a large ventral gland, and long, narrow gubernacular apophyses. Sabatieria balbutiens sp. n. is characterised by having a dorsally-directed mouth opening and assymetrical mouth cavity and spicules with small distal hook. Sabatieria pumila sp. n. is characterised by the combination of short body length, short cephalic setae, spicules with distal hook and short distal lamella, and 8-9 precloacal supplements. Sabatieria bathycopia sp. n. is characterised by having a stout body, short cephalic setae, curved gubernaculum, and spicules with swollen, lightly cuticularised distal tip.


The effects of hormone replacement therapy and raloxifene on C-reactive protein and homocysteine in healthy postmenopausal women: a randomized, controlled trial.
C-Reactive protein and homocysteine are independent risk factors for the development of cardiovascular disease. This study compared the effects of hormone replacement therapy (HRT) and raloxifene on serum C-reactive protein and homocysteine levels as markers of cardiovascular risk in healthy postmenopausal women. Healthy postmenopausal women (n = 390) were enrolled in a double blind, randomized, placebo-controlled, 6-month trial at eight out-patient sites in the United States. Women were randomly assigned to receive continuous combined HRT (0.625 mg/day conjugated equine estrogen and 2.5 mg/day medroxyprogesterone acetate), raloxifene (60 or 120 mg/day), or placebo for 6 months. C-Reactive protein and homocysteine were measured in baseline and 6-month serum samples. HRT increased C-reactive protein levels by 84% (P<0.001), whereas raloxifene (60 and 120 mg/day) had no significant effect (-6% and -4%;, respectively; P>0.2). Raloxifene (60 and 120 mg/day) significantly lowered serum levels ofhomocysteine by 8% (P = 0.014) and 6% (P = 0.024), respectively, similar to the 7% (P = 0.014) reduction obtained with HRT. We conclude that HRT and raloxifene lower serum homocysteine levels to a comparable extent in postmenopausal women. Whereas cardiovascular risk predicted by C-reactive protein in healthy postmenopausal women is not influenced by raloxifene, the relationship between elevated C-reactive protein levels with HRT and cardiovascular disease events requires further study.


Central neurotranspeptide, alpha-melanocyte-stimulating hormone (alpha-MSH) is upregulated in patients with congestive heart failure.
Alpha-melanocyte-stimulating hormone (alpha-MSH), a pro-opiomelanocortin (POMC) derivative, is a neuropeptide with potent anti-inflammatory properties that inhibits tissue injury in a wide array of inflammation models. To determine if alpha-MSH is involved in the development of congestive heart failure (CHF) with the specific aim of examining its peripheral source and one of the mechanisms. The circulating levels of alpha-MSH were measured in 115 patients with CHF using a double-antibody radioimmunoassay. To determine one of the sources of circulating alpha-MSH, human peripheral blood mononuclear cells (PBMC) were stimulated with lipopolysaccharide (LPS) or tumor necrosis factor (TNF)-alpha. Furthermore, to clarify one of the functions of alpha-MSH, PBMC were cultured in the presence or absence of alpha-MSH. Plasma levels of alpha-MSH were significantly higher in NYHA class II patients with CHF than in control subjects (p<0.0001). A significant correlation was found between the levels of alpha-MSH and high-sensitive testing for C-reactive protein in patients with CHF (r=0.41, p<0.0005). PBMC stimulated with LPS or TNF-alpha released alpha-MSH in a concentration-dependent manner. alpha-MSH inhibited LPS-induced TNF-alpha production, and alpha-MSH simultaneously augmented production of interleukin (IL)-10 by PBMC. Circulating alpha-MSH was increased in patients with CHF. Inflammatory response induced alpha-MSH production in cultured human PBMC. Treatment of alpha-MSH could modify the immunobalance between inflammatory and anti-inflammatory responses in cultured PBMC. These findings suggest that alpha-MSH may play an important role in the pathophysiology of CHF.


The Role of Neuroglial Crosstalk and Synaptic Plasticity-Mediated Central Sensitization in Acupuncture Analgesia.
Although pain is regarded as a global public health priority, analgesic therapy remains a significant challenge. Pain is a hypersensitivity state caused by peripheral and central sensitization, with the latter considered the culprit for chronic pain. This study summarizes the pathogenesis of central sensitization from the perspective of neuroglial crosstalk and synaptic plasticity and underlines the related analgesic mechanisms of acupuncture. Central sensitization is modulated by the neurotransmitters and neuropeptides involved in the ascending excitatory pathway and the descending pain modulatory system. Acupuncture analgesia is associated with downregulating glutamate in the ascending excitatory pathway and upregulating opioids, 𝛾-aminobutyric acid, norepinephrine, and 5-hydroxytryptamine in the descending pain modulatory system. Furthermore, it is increasingly appreciated that neurotransmitters, cytokines, and chemokines are implicated in neuroglial crosstalk and associated plasticity, thus contributing to central sensitization. Acupuncture produces its analgesic action by inhibiting cytokines, such as interleukin-1β, interleukin-6, and tumor necrosis factor-α, and upregulating interleukin-10, as well as modulating chemokines and their receptors such as CX3CL1/CX3CR1, CXCL12/CXCR4, CCL2/CCR2, and CXCL1/CXCR2. These factors are regulated by acupuncture through the activation of multiple signaling pathways, including mitogen-activated protein kinase signaling (e.g., the p38, extracellular signal-regulated kinases, and c-Jun-N-terminal kinase pathways), which contribute to the activation of nociceptive neurons. However, the responses of chemokines to acupuncture vary among the types of pain models, acupuncture methods, and stimulation parameters. Thus, the exact mechanisms require future clarification. Taken together, inhibition of central sensitization modulated by neuroglial plasticity is central in acupuncture analgesia, providing a novel insight for the clinical application of acupuncture analgesia.


Self-reported subjective sleep quality and fatigue in patients with peritoneal dialysis treatment at home.
The aim of this study was to describe habitual sleep, daytime symptoms, sleep-disturbing factors, current sleep during 1 week and fatigue in patients with peritoneal dialysis treatment at home and also discover predictions for sleep quality outcome. The knowledge should increase possibilities for supportive nursing health care. Fifty-five patients answered two mailed questionnaires and filled in a sleep diary. Of these, 60% had moderate, persistent sleep problems combined with daytime symptoms. Nocturnal awakenings with difficulties falling asleep again and a sleep duration predicted as 57% of sleep quality. Nocturnal pruritus and 'difficulties finding a comfortable sleeping position' were significant sleep-disturbing factors. Sleep quality predictors means progress in knowledge about the complexity of the situation for peritoneal dialysis patients. Poor sleep, daytime symptoms, sleep-disturbing factors and chronic fatigue need to be enlightened, especially for the nephrology nurses who are in a unique position to give supportive nursing health care.


[System of cAMP-dependent phosphorylation in NIH 3T3 cells passing into the resting state].
The changes in the cAMP level and the activity of cAMP-dependent protein kinase in NIH 3T3 cells going into the resting state induced by cell cultivation in a medium with 0.5% serum were investigated. It was shown that under conditions when cells are going into the resting state, the slight increase in the intracellular concentration of cAMP observed within the first 24 hours after cell transfer onto a serum-deficient medium is correlated with a simultaneous decrease of the cAMP phosphodiesterase activity. A procedure was developed allowing to measure the activity of cAMP-dependent protein kinase in cell lysates in the presence of cAMP-independent protein kinases. The activity of cAMP-dependent protein kinase in the cells going into the resting state markedly increases (up to 5-fold) after 9 days as compared to that of dividing cells. This rise in activity is due to the decrease of the content of a thermostable protein inhibitor of cAMP-dependent protein kinase rather than to the elevated level of cAMP.