Single isomer cyclodextrins as chiral selectors in capillary electrophoresis.
Since decades, cyclodextrins are one of the most powerful selectors in chiral capillary electrophoresis for the enantioseparation of diverse organic compounds. This review concerns papers published over the last decade (from 2009 until nowadays), dealing with the capillary electrophoretic application of single isomer cyclodextrin derivatives in chiral separations. Following a brief overview of their synthetic approaches, the inventory of the neutral, negatively and positively charged (including both permanently ionic and pH-tunable ionizable substituents) and zwitterionic CD derivatives is presented, with insights to underlying structural aspects by NMR spectroscopy and molecular modeling. CE represents an ideal tool to study the weak, non-covalent supramolecular interactions. The published methods are reviewed in the light of enantioselectivity, enantiomer migration order and the fine-tuning of enantiodiscrimination by the substitution pattern of the single entity selector molecules, which is hardly possible for their randomly substituted counterparts. All the reviewed publications herein support that cyclodextrin-based chiral capillary electrophoresis seems to remain a popular choice in pharmaceutical and biomedical analysis.


Crystal structure of a TFIIB-TBP-TATA-element ternary complex.
The crystal structure of the transcription factor IIB (TFIIB)/TATA box-binding protein (TBP)/TATA-element ternary complex is described at 2.7 A resolution. Core TFIIB resembles cyclin A, and recognizes the preformed TBP-DNA complex through protein-protein and protein-DNA interactions. The amino-terminal domain of core TFIIB forms the downstream surface of the ternary complex, where it could fix the transcription start site. The remaining surfaces of TBP and the TFIIB can interact with TBP-associated factors, other class II initiation factors, and transcriptional activators and coactivators.


Cromolyn sodium in the management of systemic mastocytosis.
A multicenter, double-blind, placebo-controlled trial of the efficacy of oral cromolyn sodium (200 mg orally four times per day) was conducted in 11 patients with systemic mastocytosis who had been maintained with the drug on an individualized compassionate-need basis. Efficacy was measured by physician assessment of overall disease severity based on history and physical examination at specified intervals and by the average daily patient symptom diary scores for each of three mastocytosis-related symptoms that had previously appeared to be alleviated by the use of this drug. Efficacy variables were compared for a 4-week baseline period, during which patients received open-labeled cromolyn sodium, and at 4-week intervals during a 16-week period of random assignment to cromolyn sodium or placebo. Overall disease severity and symptoms recorded in patient diaries were graded on a scale of 0 (absent) to 5 (incapacitating). The average physician assessment of disease severity and symptom scores of the patients in the placebo-treated group increased significantly during the randomization phase relative to patients in the cromolyn sodium-treated group, reflecting an exacerbation of symptoms with drug withdrawal (p less than 0.05 and less than 0.028, respectively). When the symptom scores were analyzed separately for gastrointestinal manifestations of disease (diarrhea, abdominal pain, nausea, and vomiting), cromolyn sodium treatment was significantly beneficial relative to placebo (p less than 0.02), whereas the benefit for nongastrointestinal manifestations did not reach statistical significance.


Presynaptic alteration of [3H]GABA transport in hippocampus by amygdala kindling.
gamma-Amino[2,3-3H]butyric acid ([3H]GABA) transport was investigated in hippocampal synaptosomes prepared from amygdala kindled rats. Kindling enhanced [3H]GABA uptake by 10-20%, while the Km of uptake and the kinetics of the release were unchanged. The dose response curve of the inhibition of [3H]GABA uptake by veratridine was altered by kindling: the K0.5 of inhibition and the Hill number increased and the curve became biphasic after a long period of stimulations. This effect was interpreted in terms of a modification of the functioning of the presynaptic fast Na+ channels which initially become more resistant to depolarization. In the long term, it appears that new Na+ channels develop that are more sensitive to the action of veratridine.


5'-Terminal caps of snRNAs are reactive with antibodies specific for 2,2,7-trimethylguanosine in whole cells and nuclear matrices. Double-label immunofluorescent studies with anti-m3G antibodies and with anti-RNP and anti-Sm autoantibodies.
Antibodies specific for 2,2,7-trimethylguanosine (m3G), which do not cross-react with m7G-capped RNA molecules were used to study, by immunofluorescence microscopy, the reactivity of the m3G-containing cap structures of the snRNAs U1 to U5 in situ. In interphase cells, immunofluorescent sites were restricted to the nucleus, whilst nucleoli were free of fluorescence. This indicates that the 5' terminal of most of the nucleoplasmic snRNAs are not protected by an m3G cap-recognizing protein and that the snRNA caps are not necessarily required for the binding of snRNPs to subnuclear structures. The snRNAs in the nucleoplasm appeared as distinct units in the light microscope, and this allowed the comparison of the distribution of snRNP proteins by double label studies with anti-RNP or anti-Sm antibodies within the same cell. The three antibody classes produced superimposable fluorescent patterns. Taking into account that the various IgGs react with antigenic sites on snRNAs or snRNP proteins not shared by all the snRNP species, these data suggest that U1 snRNP particles are distributed in the same way as the other snRNPs in the nucleus. Qualitatively the same results were obtained with DNase-treated nuclear matrices indicating that intact snRNPs are part of the nuclear matrix. Our data are consistent with proposals that the various snRNPs may be involved in processing of hnRNA and that this may take place at the nuclear matrix.


Stereoselective synthesis of monomeric flavonoids.
Polyphenolic compounds have recently attracted considerable interest in the field of nutrition, health and medicine. This is the result of the growing body of evidence suggesting that these compounds may act as potent antioxidants and/or modulate key biological pathways in vivo in mammals. Studies aimed at comprehending the intricate principles that govern the chemistry of these important natural products have thus accelerated over the past decade. Prominent amongst these is the ability to synthesize monomeric prototypes with and without 13C- and radio-labeling. Endeavors exploiting the stereoselective syntheses of representative classes of flavonoid monomers are reviewed here.


Aberrations of holokinetic chromosomes and associated lethality after x-irradiation of meiotic stages in Tetranychus urticae Koch (acari, tetranychidae).
Chromosomes of the holokinetic organization type were irradiated with X-rays in various stages of meiosis in unfertilized eggs of Tetranychus urticae Koch. Visible cytological aberrations, lethality and sterility were investigated in subsequent generations. Chromosome fragments are the most frequently occurring light-microscopically visible chromosome aberrations; bridges are not formed. Contrary to expectations, the presence of fragments appears to be positively correlated with the occurrence of lethality; loss of fragments, mis-segregation and the measure of damage to the broken chromosome parts are involved. In contrast with monokinetic chromosomes the earliest lethality occurs only after about 10 divisions. The ratios between different embryonic lethality types (early vs. late) differ depending on the stage irradiated: in more compact chromatin, more serious damage (i.e. more early lethality syndromes) is induced than in less compact chromatin. In the progeny of the surviving males, neither translocations nor independent fragments are found; indirect evidence indicated the occasional presence of inversions. The presumptive inversions are induced more frequently in a chromatin-compact stage (metaphase I) than in a less compact one (telophase I).


Peroxisomal ABC transporters: structure, function and role in disease.
ATP-binding cassette (ABC) transporters belong to one of the largest families of membrane proteins, and are present in almost all living organisms from eubacteria to mammals. They exist on plasma membranes and intracellular compartments such as the mitochondria, peroxisomes, endoplasmic reticulum, Golgi apparatus and lysosomes, and mediate the active transport of a wide variety of substrates in a variety of different cellular processes. These include the transport of amino acids, polysaccharides, peptides, lipids and xenobiotics, including drugs and toxins. Three ABC transporters belonging to subfamily D have been identified in mammalian peroxisomes. The ABC transporters are half-size and assemble mostly as a homodimer after posttranslational transport to peroxisomal membranes. ABCD1/ALDP and ABCD2/ALDRP are suggested to be involved in the transport of very long chain acyl-CoA with differences in substrate specificity, and ABCD3/PMP70 is involved in the transport of long and branched chain acyl-CoA. ABCD1 is known to be responsible for X-linked adrenoleukodystrophy (X-ALD), an inborn error of peroxisomal β-oxidation of very long chain fatty acids. Here, we summarize recent advances and important points in our advancing understanding of how these ABC transporters target and assemble to peroxisomal membranes and perform their functions in physiological and pathological processes, including the neurodegenerative disease, X-ALD.


Protein kinase C-mediated regulation of the expression of CD14 and CD11/CD18 in U937 cells.
We used the U937 cell line to analyze CD14, CD11/CD18, HLA class-I and DR antigen expression during PMA-induced differentiation. Treatment of U937 cells with PMA markedly increased CD14, CD11a, CD11b and CD18 antigen expression, and slightly increased CD11c expression. Protein kinase C may play a major role in regulating the expression of these antigens. The protein kinase inhibitor H7 abrogated the inductive effect of PMA. Calcium ionophore, when added alone or in the presence of PMA, had no effect. The inhibitory effect of the calcium antagonist verapamil, EGTA, and of chlorpromazine, an antagonist of calcium-binding proteins, supports a role for calcium-dependent protein kinase C in the up-regulation of CD14 and CD11/CD18 surface expression. The specific calmodulin inhibitors R24571 and W7 had no effect on antigen expression. Our findings suggest that protein kinase C activation is an important step in the PMA-induced differentiation of U937 cells.


Oliceridine is Associated with Reduced Risk of Vomiting and Need for Rescue Antiemetics Compared to Morphine: Exploratory Analysis from Two Phase 3 Randomized Placebo and Active Controlled Trials.
Use of parenteral opioids is a major risk factor for postoperative nausea and vomiting. Conventional opioids bind to µ-opioid receptors (MOR), stimulate both the G-protein signaling (achieving analgesia); and the β-arrestin pathway (associated with opioid-related adverse effects). Oliceridine, a next-generation IV opioid, is a G-protein selective MOR agonist, with limited recruitment of β-arrestin. In two randomized, placebo- and morphine-controlled phase 3 studies of patients with moderate-to-severe acute pain following bunionectomy or abdominoplasty, oliceridine at demand doses of 0.1, 0.35, and 0.5 mg provided rapid and sustained analgesia vs. placebo with favorable gastrointestinal (GI) tolerability. In this exploratory analysis, we utilized a clinical endpoint assessing gastrointestinal tolerability, "complete GI response" defined as the proportion of patients with no vomiting and no use of rescue antiemetic to characterize the GI tolerability profile of oliceridine vs. morphine. A logistic regression model was utilized to compare oliceridine (pooled regimens) vs. morphine, after controlling for analgesia (using the sum of pain intensity difference [SPID]-48/24 [bunionectomy/abdominoplasty] with pre-rescue scores carried forward for 6 h). This analysis excluded patients receiving placebo and was performed for each study separately and for pooled data from both studies. In the unadjusted analysis, a significantly greater proportion of patients in the placebo (76.4%), oliceridine 0.1 mg (68.0%), and 0.35 mg (46.2%) demand dose achieved complete GI response vs. morphine 1 mg (30.8%), p ≤ 0.005. In the adjusted analysis, after controlling for analgesia, the odds ratio of experiencing a complete GI response with oliceridine (pooled regimens) vs. morphine was 3.14 (95% CI: 1.78, 5.56; p < 0.0001) in bunionectomy study and 1.92 (95% CI: 1.09, 3.36; p = 0.024) in abdominoplasty study. When controlled for the analgesic effects (constant SPID-48/24), the odds ratio for complete GI response was higher with oliceridine than morphine, suggesting better GI tolerability with oliceridine.


An improved HPLC method for determination of carotenoids in human serum.
An HPLC method was developed to determine the various carotenoids in human serum. A C-30 column and a mobile phase of 100% methanol (A) and 100% methylene chloride (B) with the following gradient elution were used: 90% A and 10% B in the beginning, maintained for 5 min, decreased to 78% A at 15 min, 62% A at 30 min, 52% A at 40 min, 41% A at 50 min, 38% A at 55 min, maintained for 3 min, and returned to 100% A at 65 min. A total of 21 carotenoids, including all-trans forms of lutein, zeaxanthin, alpha-cryptoxanthin, beta-cryptoxanthin, alpha-carotene, beta-carotene and lycopene, as well as their 14 cis-isomers were resolved within 51 min at a flow rate of 1.0 mL/min and detection at 476 nm. all-trans-beta-Carotene was found to be present in highest amount (256.3-864.2 ng/mL), followed by all-trans-lycopene (64.4-569.2 ng/mL), all-trans-lutein (137.9-450.3 ng/mL), all-trans-alpha-cryptoxanthin (55.7-188.2 ng/mL), all-trans-beta-cryptoxanthin (43.1-134.5 ng/mL), all-trans-alpha-carotene (20.0-122.1 ng/mL) and all-trans-zeaxanthin (9.1-21.3 ng/mL). Similar trend was observed for cis-isomers of carotenoids.


Effects of a long-term high-fat diet and switching from a high-fat to low-fat, standard diet on hepatic fat accumulation in Sprague-Dawley rats.
To investigate the effects of a long-term high-fat diet and switching from high-fat to a low-fat diet on hepatic fat accumulation in Sprague-Dawley (SD) rats, 3-week-old male SD rats were fed a high-fat diet (HFD) containing 45% fat (kilocalories) for 43 weeks (HDHD group), an HFD for 23 weeks followed by a low-fat, standard diet (LFD) containing 10% fat for 20 weeks (HDLD group), and an LFD for 43 weeks (LDLD group). Histopathologically, steatosis and lobular inflammation was obvious in the HDLD and HDHD groups at 46 weeks of age, and ballooning hepatocytes and Mallory hyalines were seen in the HDHD group. Mild fibrosis was observed in 5 of 13 (38%) rats in the HDHD or HDLD groups. Our results demonstrate that a long-term high-fat diet can induce nonalcoholic steatohepatitis (NASH) in SD rats. Switching to a low-fat, standard diet prevented the progression of NASH, although steatosis was not improved.


Carcinoid tumor of the spleen: report of a case.
Carcinoid tumors are uncommon tumors of the neuroendocrine system. They grow slowly and may remain silent for years before presenting with carcinoid syndrome. A diagnosis of asymptomatic carcinoid tumor is difficult. Wide resection of the primary tumor and metastatic lesions is the first choice of treatment. Primary carcinoid is sometimes distributed throughout the entire body, but it is rare in the spleen. We herein present a rare case of a symptomless carcinoid tumor that predominantly invaded the spleen with liver metastasis.


Prevalence and impact of malnutrition on outcomes and mortality of under-five years children with pneumonia: a study from Upper Egypt.
Malnutrition has adverse impacts on under-five children with pneumonia. The purpose of this study was to address the prevalence and impact of malnutrition on under-five years children with pneumonia, admitted to a tertiary large children hospital in Upper Egypt. This study is a prospective case-control study. All under-five children diagnosed with pneumonia who were admitted to Assiut University Children's Hospital (AUCH) from January 1st to December 31st, 2021, were enrolled. Based on their nutritional assessment, the studied participants were classified into 2 groups: (1): Children with pneumonia and with nutritional deficiency considered as cases, and (2): Children with pneumonia and without nutritional deficiency considered as controls. Three hundred-fifty cases and 154 control subjects were enrolled, respectively. 93.4%, 31.1%, and 61.7% of the cases had underweight, stunting, and wasting, respectively. Among those cases, there were significant differences between survivors and non-survivors with regard to some clinicodemographic factors, laboratory parameters, and anthropometric parameters. Lack of compulsory vaccination, presence of sepsis, and blood transfusion (OR 2.874, 95% CI 0.048 - 2.988, p = 0.004, 2.627, 0.040 - 2.677, p = 0.009, and 4.108, 0.134 - 3.381, p < 0.001, respectively) were significant independent predictors for mortality among malnourished children with pneumonia. Conclusion: Malnutrition has a high prevalence in under-five children with pneumonia in our locality. It has adverse effects on the outcomes and in-hospital mortality of those children. Lack of compulsory vaccination, presence of sepsis, and blood transfusion were significant independent predictors of mortality in malnourished children with pneumonia. Larger multicenter studies are warranted. What is Known: • Malnutrition has adverse impacts on under-five children with pneumonia. • Malnutrition could be a reason for in-hospital mortality among under-five children with pneumonia. What is New: • Malnutrition has a high prevalence in under-five children with pneumonia in Upper Egypt, with its adverse effects on the outcomes and mortality of those children. • Lack of vaccination, presence of sepsis, and blood transfusion are significant independent predictors of mortality in malnourished children with pneumonia in Upper Egypt.


Simultaneous electroencephalographic and functional magnetic resonance imaging indicate impaired cortical top-down processing in association with anesthetic-induced unconsciousness.
In imaging functional connectivity (FC) analyses of the resting brain, alterations of FC during unconsciousness have been reported. These results are in accordance with recent electroencephalographic studies observing impaired top-down processing during anesthesia. In this study, simultaneous records of functional magnetic resonance imaging (fMRI) and electroencephalogram were performed to investigate the causality of neural mechanisms during propofol-induced loss of consciousness by correlating FC in fMRI and directional connectivity (DC) in electroencephalogram. Resting-state 63-channel electroencephalogram and blood oxygen level-dependent 3-Tesla fMRI of 15 healthy subjects were simultaneously registered during consciousness and propofol-induced loss of consciousness. To indicate DC, electroencephalographic symbolic transfer entropy was applied as a nonlinear measure of mutual interdependencies between underlying physiological processes. The relationship between FC of resting-state networks of the brain (z values) and DC was analyzed by a partial correlation. Independent component analyses of resting-state fMRI showed decreased FC in frontoparietal default networks during unconsciousness, whereas FC in primary sensory networks increased. DC indicated a decline in frontal-parietal (area under the receiver characteristic curve, 0.92; 95% CI, 0.68-1.00) and frontooccipital (0.82; 0.53-1.00) feedback DC (P<0.05 corrected). The changes of FC in the anterior default network correlated with the changes of DC in frontal-parietal (rpartial=+0.62; P=0.030) and frontal-occipital (+0.63; 0.048) electroencephalographic electrodes (P<0.05 corrected). The simultaneous propofol-induced suppression of frontal feedback connectivity in the electroencephalogram and of frontoparietal FC in the fMRI indicates a fundamental role of top-down processing for consciousness.


Sharing the SAGA.
Transcription initiation is a major regulatory step in eukaryotic gene expression. Co-activators establish transcriptionally competent promoter architectures and chromatin signatures to allow the formation of the pre-initiation complex (PIC), comprising RNA polymerase II (Pol II) and general transcription factors (GTFs). Many GTFs and co-activators are multisubunit complexes, in which individual components are organized into functional modules carrying specific activities. Recent advances in affinity purification and mass spectrometry analyses have revealed that these complexes often share functional modules, rather than containing unique components. This observation appears remarkably prevalent for chromatin-modifying and remodeling complexes. Here, we use the modular organization of the evolutionary conserved Spt-Ada-Gcn5 acetyltransferase (SAGA) complex as a paradigm to illustrate how co-activators share and combine a relatively limited set of functional tools.


Photocatalytic hydrogen production by water/methanol decomposition using Au/TiO2 prepared by deposition-precipitation with urea.
Gold nanoparticles deposited on TiO2 Degussa P25, prepared by deposition-precipitation with urea, were studied in the photocatalytic hydrogen production. The effect of parameters such as mass of catalyst, gold loading, thermal treatment, and atmosphere of treatment was evaluated and optimized. The presence of metallic gold on the titania surface showed to have contributed to the high improvement in the activity of bare TiO2 for hydrogen generation under UV light (λ=254 nm) using a lamp of low energy (2W) consumption. The optimal gold loading for the photocatalysts was 0.5 wt.%, the mass of catalyst in the reactor was 0.5 g/L in a water/methanol 1:1 vol. solution, and the thermal treatment that produced the most active gold nanoparticles was found at 300°C. The photocatalysts thermally treated under hydrogen at 300°C produced 1492 μmol g(-1)h(-1) of hydrogen; the same catalyst activated in air produced 1866 μmo lg(-1)h(-1) of hydrogen.


Graphene oxide leads to mitochondrial-dependent apoptosis by activating ROS-p53-mPTP pathway in intestinal cells.
Owing to its unique physical and chemical properties, graphene oxide (GO) has a wide range of applications in biomedical field. However, with the gradual improvement of biosafety investigations on nanomaterials, growing literatures have pointed out that GO could lead to oxidative stress, aggravation of inflammatory responses, and even irreversible lesions in human multi-tissues, while its damage to small intestinal remained unclear. In this study, we conducted an in-depth study on the toxicological effect of GO on intestinal tissues, and further clarified its toxic effect and molecular mechanism on inducing intestinal cell death. Firstly, we characterized the shape size, potential value, Fourier Transform infrared spectroscopy (FT-IR) characterization and pro-oxidant properties of GO nanosheets. The cytotoxicity of different concentrations of GO to Caco-2 and IEC-6 cell lines was thereafter observed, which was specifically manifested as invoking NADPH Oxidase 1 (NOX1) proteins, accompanied generation of reactive oxygen species (ROS). Since that, more p53 flowed into mitochondria to combine with cyclophilin D (CYPD), thus induced mitochondrial permeability transition pore (mPTP) opening. Through ROS-CyPD-mPTP signaling pathway, GO exerted imbalance of mitochondrial homeostasis, while released cytochrome c (CytC) would ultimate caspase-dependent cell apoptosis. In vivo experiment also confirmed that the microstructure of small intestine was damaged, and the apoptosis rate and oxidative markers were significantly increased in GO-treated Sprague- Dawley (SD) rats (40 mg/kg once every other day from day 1 to day 9 by oral gavage). Based on these findings, we conclude that the adverse effects of oral exposure of GO on the biological system mainly concentrate in the digestive tract, and clarify the key role of ROS-mitochondrial homeostasis-apoptosis axis in GO-derived intestinal toxicity. Considering all these results and the fact that GO exhibited intestinal toxicity, we believe that this research providing a safety reference for its biomedical applications.


Comparison of acellular pertussis vaccines-induced immunity against infection due to Bordetella pertussis variant isolates in a mouse model.
A significant increase in the incidence of pertussis in adolescents and adults has been observed in vaccinated populations. Concomitantly, emergence of novel pertussis toxin and pertactin types in circulating Bordetella pertussis isolates was noticed. In this study, immunity induced by acellular vaccines against infection due to isolates expressing different pertactin types and fimbriae was monitored in a mouse model. In accordance with previous studies, the effect of a bicomponent DTPa vaccine on bacterial clearance was lower when compared with tri- or pentavalent DTPa vaccines. Whatever the isolates used to infect mice, the tri- or pentavalent DTPa vaccines were both efficacious in inducing immunity that resulted in clearance of infection. These findings suggest that re-emergence of pertussis might not be related to emergence of isolates escaping vaccine protection. The present study reduces potential concerns about acellular vaccine efficacy, but frequent monitoring of protection and surveillance of the evolution of the B. pertussis population remains of particular importance.


Arachnoid Cyst-Associated Chronic Subdural Hematoma: Report of 14 Cases and a Systematic Literature Review.
Arachnoid cyst (AC)-associated chronic subdural hematoma (CSDH) differs significantly from its counterparts without AC in epidemiologic, demographic, and clinical characteristics, as well as in management and prognosis. This study was conducted to further examine the epidemiologic, demographic, and clinical characteristics; diagnosis; treatment; and prognosis of AC-associated CSDH. This was a retrospective study of the medical records at the neurosurgical departments of 2 institutions along with a systematic PubMed search for relevant studies published in English or Chinese. A total of 182 patients (148 males; 81.3%) were evaluated, including 14 cases in our present series. The patients ranged in age from 1 to 80 years (mean age, 24.41 ± 13.69 years). Among the 175 patients with adequate prehospital history information, 119 (68%) had a history of recent head trauma or sport-related injury. AC locations included the middle fossa and sylvian fissure in 162 cases (89.0%), cerebral convexity in 17 cases (9.3%), posterior fossa in 2 cases (1.1%), and interhemispheric fissure in 1 case (0.5%). Among the 161 patients with specific data on outcomes, 159 (98.8%) had favorable recovery, 1 patient had an evident neurologic deficit, and 1 patient died from cardiac arrest. CSDH is a rare complication in patients with intracranial AC. Male children, juveniles, and young adults with recent head trauma or sport-related injury are most commonly inflicted. Burr hole drainage is the first-choice surgical procedure in symptomatic patients and is still effective in some recurrent cases. Fenestration or resection of the AC membrane is not a requisite in patients with previous asymptomatic AC.


New generation of wearable goniometers for motion capture systems.
Monitoring joint angles through wearable systems enables human posture and gesture to be reconstructed as a support for physical rehabilitation both in clinics and at the patient's home. A new generation of wearable goniometers based on knitted piezoresistive fabric (KPF) technology is presented. KPF single-and double-layer devices were designed and characterized under stretching and bending to work as strain sensors and goniometers. The theoretical working principle and the derived electromechanical model, previously proved for carbon elastomer sensors, were generalized to KPF. The devices were used to correlate angles and piezoresistive fabric behaviour, to highlight the differences in terms of performance between the single layer and the double layer sensors. A fast calibration procedure is also proposed. The proposed device was tested both in static and dynamic conditions in comparison with standard electrogoniometers and inertial measurement units respectively. KPF goniometer capabilities in angle detection were experimentally proved and a discussion of the device measurement errors of is provided. The paper concludes with an analysis of sensor accuracy and hysteresis reduction in particular configurations. Double layer KPF goniometers showed a promising performance in terms of angle measurements both in quasi-static and dynamic working mode for velocities typical of human movement. A further approach consisting of a combination of multiple sensors to increase accuracy via sensor fusion technique has been presented.


One step construction of an electrochemical sensor for melamine detection in milk towards an integrated portable system.
Excessive intake of melamine (MEL) can be harmful to human health, and it is important to establish a rapid and accurate MEL detection method. As the electrochemical activity of MEL is very low, ferrocenylglutathione (Fc-ECG) was used as an electron transfer mediator to assist with the detection of MEL using screen-printed carbon electrode (SPCE). This modified electrode (MEL/Fc-ECG/SPCE) was prepared by stepwise drop-casting and was fully characterized. Results showed that MEL significantly enhanced signal of Fc-ECG/SPCE sensor due to the three p-π conjugated double bonds that facilitated electron transfer. Under optimal conditions, the sensor exhibits two linearities in the range of 0.20-2.00 μM and 8.00-800 μM, with a sensitivity of 15.03 μA·μM-1·cm-2. The selectivity, stability, and reproducibility were investigated and successfully used to detect MEL in raw milk and confirms safety verification of foods. Moreover, a portable testing platform was designed for MEL detection based on a CH32 chip.


Heterogeneity among membrane vesicles of Escherichia coli: effects of production and fractionation techniques.
We have previously shown that Escherichia coli membrane proteins are associated with subpopulations of membrane vesicles beyond the well-defined inner versus outer membrane classification. To separate these vesicles we used a shallow sucrose gradient which differed in many respects from established procedures. Here we compare this revised technique to the classical sucrose density centrifugation procedure. We found that certain manipulations common to the latter obscured heterogeneity among membrane vesicles. The following treatments degraded vesicle separation: growth in rich medium addition of EDTA to buffers, spheroplasting, sonication, pelleting of membranes prior to sucrose gradient centrifugation, overloading the gradient, and long centrifugation times. Some treatments, such as EDTA addition, affected only selected vesicles. When determining protein localization within bacterial membranes experiments should be designed to avoid or at least minimize these manipulations.


Epigenetic mechanisms within the sperm epigenome and their diagnostic potential.
The sperm epigenome contains a highly unique and specialized epigenetic landscape. Insightful questions need be asked about these epigenetic signatures and their predictive potential to assess the approximately 1 in 6 couples who experience infertility. Among those couples that do experience infertility, approximately half of the cases involve a male factor. Unfortunately, there is a significant lack of effective diagnostic tools in the male infertility space and thus clinicians are left with little data upon which they can formulate data driven treatment plans. Taking together this information and the striking prevalence of male infertility it's obvious that there is a need for improved diagnostic techniques for male infertility. Many studies have identified what appear to be clinically meaningful epigenetic alterations in sperm that may add utility in the diagnoses of infertility and improvement of pregnancy outcomes. Many researchers believe that continued analysis of these various epigenetic mechanisms may provide powerful predictive insight. In fact, there is promising current data suggesting that the predictive power of DNA methylation, Nuclear Proteins, and miRNA signatures in sperm likely can improve what is currently found with traditional diagnosis of male infertility. The focus of this review is to give a brief understanding to the field of epigenetics and the potential predictive power the sperm epigenome may hold in relation to improving the treatment and diagnosis of male infertility patients.


Dopamine D2 receptor-mediated epidermal growth factor receptor transactivation through a disintegrin and metalloprotease regulates dopaminergic neuron development via extracellular signal-related kinase activation.
Dopamine D2 receptor (D2R)-mediated extracellular signal-regulated kinase (ERK) activation plays an important role in the development of dopaminergic mesencephalic neurons. Here, we demonstrate that D2R induces the shedding of heparin-binding epidermal growth factor (EGF) through the activation of a disintegrin and metalloprotease (ADAM) 10 or 17, leading to EGF receptor transactivation, downstream ERK activation, and ultimately an increase in the number of dopaminergic neurons and their neurite length in primary mesencephalic cultures from wild-type mice. These outcomes, however, were not observed in cultures from D2R knock-out mice. Our findings show that D2R-mediated ERK activation regulates mesencephalic dopaminergic neuron development via EGF receptor transactivation through ADAM10/17.


Effects of foot placement on postural stability of construction workers on stilts.
Stilts are elevated tools that are frequently used by construction workers to raise workers 18-40 inches above the ground. The objective of this laboratory study was to evaluate the potential loss of postural stability associated with the use of stilts in various foot placements. Twenty construction workers with at least 1 year of experience in the use of stilts participated in this study. One Kistler force platform was used to collect kinetic data. Participants were tested under six-foot-placement conditions. These 6 experimental conditions were statically tested under all combinations of 3 levels of elevation: 0'' (no stilts), 24'' stilt height and 40'' stilt height. SAS mixed procedure was used to evaluate the effect of different experimental conditions. The results of the multivariate analysis of variance (MANOVA) and repeated measures of univariate analyses of variance (ANOVAs) demonstrated that stilt height, foot-placement direction, and foot-placement width all had significant effects on the whole-body postural stability. This study found that the higher the stilts were elevated, the greater the postural instability. A stance position with one foot placed forward of the other foot produced greater postural instability than a position with the feet parallel and directly beneath the body. This study found that placement of the feet parallel and directly beneath the body, with the feet positioned a half shoulder width apart, caused a greater amount of postural sway and instability than one and one-and-half shoulder width. This study also found that construction workers using the stilts could perceive the likely postural instability due to the change in foot placements.


Costs of abandoned coal mine reclamation and associated recreation benefits in Ohio.
Two hundred years of coal mining in Ohio have degraded land and water resources, imposing social costs on its citizens. An interdisciplinary approach employing hydrology, geographic information systems, and a recreation visitation function model, is used to estimate the damages from upstream coal mining to lakes in Ohio. The estimated recreational damages to five of the coal-mining-impacted lakes, using dissolved sulfate as coal-mining-impact indicator, amount to $21 Million per year. Post-reclamation recreational benefits from reducing sulfate concentrations by 6.5% and 15% in the five impacted lakes were estimated to range from $1.89 to $4.92 Million per year, with a net present value ranging from $14.56 Million to $37.79 Million. A benefit costs analysis (BCA) of recreational benefits and coal mine reclamation costs provides some evidence for potential Pareto improvement by investing limited resources in reclamation projects.


Proteome reallocation from amino acid biosynthesis to ribosomes enables yeast to grow faster in rich media.
Several recent studies have shown that the concept of proteome constraint, i.e., the need for the cell to balance allocation of its proteome between different cellular processes, is essential for ensuring proper cell function. However, there have been no attempts to elucidate how cells' maximum capacity to grow depends on protein availability for different cellular processes. To experimentally address this, we cultivated Saccharomyces cerevisiae in bioreactors with or without amino acid supplementation and performed quantitative proteomics to analyze global changes in proteome allocation, during both anaerobic and aerobic growth on glucose. Analysis of the proteomic data implies that proteome mass is mainly reallocated from amino acid biosynthetic processes into translation, which enables an increased growth rate during supplementation. Similar findings were obtained from both aerobic and anaerobic cultivations. Our findings show that cells can increase their growth rate through increasing its proteome allocation toward the protein translational machinery.


Metal [100] Nanowires with Negative Poisson's Ratio.
When materials are under stretching, occurrence of lateral contraction of materials is commonly observed. This is because Poisson's ratio, the quantity describes the relationship between a lateral strain and applied strain, is positive for nearly all materials. There are some reported structures and materials having negative Poisson's ratio. However, most of them are at macroscale, and reentrant structures and rigid rotating units are the main mechanisms for their negative Poisson's ratio behavior. Here, with numerical and theoretical evidence, we show that metal [100] nanowires with asymmetric cross-sections such as rectangle or ellipse can exhibit negative Poisson's ratio behavior. Furthermore, the negative Poisson's ratio behavior can be further improved by introducing a hole inside the asymmetric nanowires. We show that the surface effect inducing the asymmetric stresses inside the nanowires is a main origin of the superior property.


Diagnostic accuracy of coronary angiography using 64-slice computed tomography in coronary artery disease.
To conduct a meta-analysis and investigate the diagnostic value of 64-slice computed tomography (CT) angiography for diagnosing coronary artery disease (CAD) in patients. A comprehensive literature search from March 2005 to August 2014 was performed on the following databases: Cochrane Library; Medline; EmBase; PubMed; and BioMed Central database. As a reference standard, studies that assessed 64-slice CT angiography in detecting coronary artery stenosis (CAS) with invasive coronary angiography were included. Coronary artery stenosis was defined as ≥50% diameter stenosis. Diagnostic value was determined by pooling sensitivity, specificity, positive likelihood ratio (PLR) and negative likelihood ratio (NLR) values at segment-level analysis. Diagnostic accuracy was undertaken using area under the curve (AUC) value and summary receiver operating characteristic (SROC) curves. Publication bias was examined by Deek's funnel plot asymmetry test. Eight studies were included in the analysis, enrolling a total of 579 patients (7,407 segment coronary vessels). At segment-level, pooled sensitivity value was 90% (95% confidence interval [CI]: 83-95%), specificity was 91% (95% CI: 61-98%), PLR value was 9.7 (95% CI: 1.8-53.3), and NLR value was 0.11 (95% CI: 0.05-0.22) for CAS. Optimal cut-off point of sensitivity was 90%, and specificity under the SROC curve was 91%. The AUC value was 0.94. The 64-slice CT angiography is a reliable tool for detection of CAD when using a cut-off of more than or equal to 50% diameter stenosis in elderly population.


Prognostic significance of anatomic subsites: results of radiation therapy for 66 patients with localized orbital marginal zone B cell lymphoma.
To evaluate the prognostic significance of anatomic subsites for marginal zone B cell lymphoma confined to the orbit. From 1995 to 2005, 66 patients (80 lesions) with marginal zone B cell lymphoma, confined to the orbit, received radiation therapy at Samsung Medical Center. The most frequent site of involvement was conjunctiva (29 patients) followed by retrobulbar (20 patients), eyelid (10 patients) and lacrimal gland (7 patients). The median follow-up duration was 50 (13-114) months. Local relapse developed in two patients, systemic relapse in three, and metachronous relapse in two. The 5-year relapse-free survival rate was 92.0%. A primary tumor located at lacrimal gland was the only poor prognostic factor for relapse (p=0.023, HR=12.263, 95% CI=1.360-110.602). The 5-year actuarial relapse-free survival rate was 57.1% for lacrimal tumor versus 95.8% for non-lacrimal tumors (p=0.002). Localized primary orbital MALT lymphomas respond extremely well to moderate-dose RT, with a high local control rate and durable disease free status. However, primary lacrimal involvement maybe related to future relapse more frequently than other orbital subsites.


Exploring the impact of exercise and essential amino acid plus cholecalciferol supplementation on physical fitness and body composition in multiple sclerosis: A case study.
In MS patients, especially those frail or malnourished, combining home-based exercise twice weekly with essential amino acids and vitamin D may improve body composition, strength, and physical performance, enabling long-term functional improvements. Multiple sclerosis (MS) is associated with reduced bone and muscle strength and function. We aimed to investigate the effectiveness of a 24-week intervention in a 57-year-old frail female with MS. The participant completed a 2×/week exercise intervention and ingested 2×/day a supplement containing 7.5 g essential amino acids and 500 IU cholecalciferol. Body composition, 6-m gait speed (GS), handgrip strength (HGS), 30-sec arm-curl test (30ACT), 6-min walking test (6MWT), 30-sec chair-stand test (30CST), and plasma concentrations of 25-hydroxyvitamin D3 [25(OH)D3], insulin-like growth factor 1 (IGF-1), and amino acids were assessed at baseline, and at Weeks 12 and 24. Plasma 25(OH)D3 increased from 23.2 to 41.3 ng/mL and IGF-1 from 131.6 to 140.7 ng/mL from baseline to post-intervention. BMI, total lean tissue mass (LTM), fat mass, bone mineral content, and the sum of 17 amino acids increased by 3.8, 1.0, 3.5, 0.2, and 19%, respectively, at Week 24. There were clinically significant increases in regional LTM (6.9% arms and 6.3% legs) and large increases in GS (67.3%), dominant HGS (31.5%), non-dominant HGS (11.8%), dominant 30ACT (100%), non-dominant 30ACT (116.7%), 6MWT (125.6%), and 30CST (44.4%). The current intervention was effective in improving components of physical fitness and body composition in a female with MS.


Successive remodeling of IgG glycans using a solid-phase enzymatic platform.
The success of glycoprotein-based drugs in various disease treatments has become widespread. Frequently, therapeutic glycoproteins exhibit a heterogeneous array of glycans that are intended to mimic human glycopatterns. While immunogenic responses to biologic drugs are uncommon, enabling exquisite control of glycosylation with minimized microheterogeneity would improve their safety, efficacy and bioavailability. Therefore, close attention has been drawn to the development of glycoengineering strategies to control the glycan structures. With the accumulation of knowledge about the glycan biosynthesis enzymes, enzymatic glycan remodeling provides a potential strategy to construct highly ordered glycans with improved efficiency and biocompatibility. In this study, we quantitatively evaluate more than 30 enzymes for glycoengineering immobilized immunoglobulin G, an impactful glycoprotein class in the pharmaceutical field. We demonstrate successive glycan remodeling in a solid-phase platform, which enabled IgG glycan harmonization into a series of complex-type N-glycoforms with high yield and efficiency while retaining native IgG binding affinity.


Targeted head CT reduction for pediatric patients with hydrocephalus and traumatic brain injury: academic center institutional experience as an example of opportunities for further improvement.
Recent studies aim to reduce radiation exposure associated with computed tomography (CT) using rapid-sequence magnetic resonance imaging (MRI). We evaluated imaging modalities used for hydrocephalus and traumatic brain injury (TBI) to identify opportunities for further radiation exposure reduction. Pediatric (≤ 18 years) patients, with either hydrocephalus or TBI receiving a head CT or head MRI from 2009 to 2017, were quantified using ICD9, ICD10, and CPT codes at a large university hospital. The odds ratios of receiving each imaging modality year-to-year and receiving a MRI or CT in a given year with each diagnosis were calculated. Beginning in 2015, hydrocephalus patients were more likely to receive a MRI vs CT (p < 0.0001), with likelihood increasing in the following 2 years. TBI patients were more likely to receive a CT than an MRI from 2009 to 2017. There was a smaller overall decrease in CT scans in TBI patients (p < 0.05) than hydrocephalus patients (p < 0.0001) and a larger increase in MRI use in hydrocephalus patients (p < 0.05) than TBI patients (p < 0.05) from 2009 to 2017. Use of CT to evaluate hydrocephalus significantly decreased over the last 3-year time period. However, CT use for TBI patients was inconsistent and did not decrease proportionally, potentially representing a belief that CT is more effective than MRI for detecting hemorrhage. There was greater use of MRI as an alternate imaging method in hydrocephalus patients than TBI patients. Head injury remains an area of improvement to decrease pediatric radiation exposure at our institution and may be an area in need of attention more broadly.


Percutaneous Transgluteal Ureteral Stump Access: A Novel Approach to Hybrid Management of Ureteral Stump Syndrome.
Duplicated collecting systems with minimal-to-no function of the upper moiety are often treated surgically with upper pole heminephrectomy and ureterectomy. Complications associated with the ureteral stump can occur and are collectively known as ureteral stump syndrome. We present a case of ureteral stump syndrome treated with percutaneous transgluteal access into the stump which aided in identification of an ectopic ureteral opening that could not be identified by surgical means alone.


Effect of short and long range order on crystal structure interpretation: Raman and powder X-ray diffraction of LiPF₆.
The structure of LiPF6 has been probed using Raman scattering as well as pXRD and the results are compared and contrasted. The conventional Bragg angle scattering pXRD determines that dry LiPF6 crystallizes in a trigonal structure (Space Group R-3 (148)), while Raman data suggests that the observed structure is close to cubic (Space Group Fm-3m (225)), similar to NaPF6 and KPF6. DFT heat capacity calculations using ab-initio methods for both R-3 and Fm-3m structures show better correlation with the Raman observations. The differences between Raman and pXRD data in LiPF6 appear to be in common with that observed in other materials that are close to phase transitions at the temperature where order/disorder and phase transition processes are known to occur. In these circumstances, Raman spectroscopy emerges as a more sensitive probe of local structural changes than pXRD, which is known to probe the overall average long range order of crystalline materials. The results are interpreted through the relationship between the local symmetry, internal energy and the heat capacity.


Occurrence and backtracking of microplastic mass loads including tire wear particles in northern Atlantic air.
Few studies report the occurrence of microplastics (MP), including tire wear particles (TWP) in the marine atmosphere, and little data is available regarding their size or sources. Here we present active air sampling devices (low- and high-volume samplers) for the evaluation of composition and MP mass loads in the marine atmosphere. Air was sampled during a research cruise along the Norwegian coast up to Bear Island. Samples were analyzed with pyrolysis-gas chromatography-mass spectrometry, generating a mass-based data set for MP in the marine atmosphere. Here we show the ubiquity of MP, even in remote Arctic areas with concentrations up to 37.5 ng m-3. Cluster of polyethylene terephthalate (max. 1.5 ng m-3) were universally present. TWP (max. 35 ng m-3) and cluster of polystyrene, polypropylene, and polyurethane (max. 1.1 ng m-3) were also detected. Atmospheric transport and dispersion models, suggested the introduction of MP into the marine atmosphere equally from sea- and land-based emissions, transforming the ocean from a sink into a source for MP.


Shear- vs. nanotopography-guided control of growth of endothelial cells on RGD-nanoparticle-nanowell arrays.
Endothelialization of therapeutic cardiovascular implants is essential for their intravascular hemocompatibility. We previously described a novel nanowell-RGD-nanoparticle ensemble, which when applied to surfaces led to enhanced endothelialization and retention under static conditions and low flow rates. In the present study we extend our work to determine the interrelated effects of flow rate and the orientation of ensemble-decorated surface arrays on the growth, adhesion and morphology of endothelial cells. Human umbilical vascular endothelial cells (HUVECs) were grown on array surfaces with either 1 μm × 5 μm spacing ("parallel to flow") and 5 μm × 1 μm spacing ("perpendicular to flow") and were exposed to a range of shear stress of (0 to 4.7 ± 0.2 dyn·cm-2 ), utilizing a pulsatile flow chamber. Under physiological flow (4.7 ± 0.2 dyn·cm-2), RGD-nanoparticle-nanowell array patterning significantly enhanced cell adhesion and spreading compared with control surfaces and with static conditions. Furthermore, improved adhesion coincided with higher alignment to surface patterning, intimating the importance of interaction and response to the array surface as a means of resisting flow detachment. Under sub-physiological condition (1.7 ± 0.3 dyn·cm-2; corresponding to early angiogenesis), nanowell-nanoparticle patterning did not provide enhanced cell growth and adhesion compared with control surfaces. However, it revealed increased alignment along the direction of flow, rather than the direction of the pattern, thus potentially indicating a threshold for cell guidance and related retention. These results could provide a cue for controlling cell growth and alignment under varying physiological conditions.


Trends in total cholesterol control among American adults with hypercholesterolemia, 1988-2018.
Cholesterol control and management in patients with hypercholesterolemia are significant for the primary and secondary prevention of atherosclerotic cardiovascular disease. This study analyzed the trend of serum total cholesterol (TC) control (<240 mg/dL and <200 mg/dL) in American adults with hypercholesterolemia and thereby make some effective recommendations for the public health measures. Basing on the National Health and Nutrition Examination Survey (NHANES) data from 1988 to 2018 (12 cycles), we calculated the weighted and representative rate of patients with hypercholesterolemia who had controlled TC, and then described the trend. Among the adults with hypercholesterolemia, the age-adjusted rate of those whose TC was less than 240 mg/dL increased from 7.67% (95%CI: 5.94%-9.40%) in 1988-1991 to 58.52% (95%CI: 55.89%-61.15%) in 2013-2014 and then remained stable; and the age-adjusted rate of those whose TC was less than 200 mg/dL increased from 2.49% (95%CI: 1.48%-3.50%) in 1988-1991 to 44.58% (95%CI: 40.00%-49.16%) in 2017-2018. We concluded that the rate of controlling TC below 200 mg/dL among all patients had shown an increasing trend from 1988 to 2018 in America, while the rate of controlling TC below 240 mg/dL remained stable in recent years after an increasing.


Frying--effect of coating on crust microstructure, color, and texture of lean meat portions.
Skinless, deboned chicken breast fillets were deep fat fried (190 °C) with and without a batter and breading coating system for up to 6 min (internal temp 70°C). Non-coated samples show surface muscle fiber splitting as soon as placed in oil. As time progressed, the depth of damage increased. Shear force and work to shear increased significantly as a dense hard crust layer was forming. By the end, fillets lost about 1/3 of their weight and had a dark, dry, brittle crust. Frying with coating protected the meat inside and no major damages to muscle fibers were observed. Shear force also increased but the values were about half of what was observed in non-coated products, meaning that the meat inside was protected. Coated products became progressively golden brown (increased a* and b*, decreased L*) while the crust adhered well to the product. After frying, the products showed a small gain in weight which was the result of no structural shrinkage and absorption of a small amount of oil.


Aerobic nitrous oxide emission in anoxic/aerobic and intermittent aeration sequencing batch reactors.
This study characterized nitrogen removal and nitrous oxide (N2O) emissions from lab-scale anoxic/aerobic sequencing batch reactor (AOSBR) and intermittent aeration sequencing batch reactor (IASBR), respectively, for treating synthetic municipal wastewater. The N2O emission was evaluated in a simulated cycle, and batch conditions of aerobic nitrification, simultaneous nitrification and denitrification (SND), and aerobic denitrification. The results show that nitrogen removal was enhanced in IASBR compared to AOSBR, with 94.2% and 67.9% of total inorganic nitrogen removal efficiency in IASBR and AOSBR, respectively. In the simulated cycle, the emission factors (of oxidized ammonium) were 4.9% and 0.6% in AOSBR and IASBR, respectively. Under batch conditions, the N2O emission factors during SND were obviously higher than that during aerobic nitrification and denitrification. The N2O emission factors during SND ranging 0.68-11.68% in AOSBR and 1.25-5.13% in IASBR. Furthermore, N2O emission under batch conditions was affected by the aeration ratios. Moderate and high aeration ratios used in this study stimulated the N2O emission from SND. The N2O emission was enhanced with the nitrite accumulation during aerobic nitrification when the nitrite-oxidizing bacteria was inhibited by a chemical inhibitor. Aerobic denitrification via nitrite could be the main pathway of N2O generation from SND processes. The findings from our study can help further understand N2O emission mechanisms and guide the optimization of the current wastewater treatment process for minimizing N2O emission.


Modern urodynamic evaluations in the urologist's office.
Patients with voiding disturbances comprise 20 per cent of the yearly outpatient visits to urologists. Of this number 25 per cent have significant neurogenic bladder dysfunction. The theoretical basis and experimental demonstration of urodynamics have been established and reported by multiple investigators during the last 15 years. Recent advances in clinical applications of fast-flow gas cystometry have led to the development of complete, portable urodynamic units that can be used conveniently in the urologist's office and are capable of simultaneously recording parameters of detrusor, urethral and skeletal muscle phincter function. This equipment is simple, efficient, economical and can be operated by well trained paramedical personnel, with minimal immediate direction required by a physician. With recent recognition by many third party carriers this equipment may amortize itself rapidly but, more importantly, it may serve to identify neurological diseases of the bladder that are undetected by other conventional methods of investigation. During the last 9 months 213 patients have undergone urodynamic evaluation at our outpatient urology clinic. These patients were categorized according to type and incidence of voiding disturbance and the number of neurogenic bladders diagnosed within each group was compiled and discussed. Several of the more interesting and unusual clinical examples are presented to familiarize the urologist with the interpretative aspects of this study.


Did a "lucky shot" sink the submarine H.L. Hunley?
The H.L. Hunley was the first submarine to be successful in combat, sinking the Union vessel Housatonic outside Charleston Harbor in 1864 during the Civil War. However, despite marking a milestone in military history, little is known about this vessel or why it sank. One popular theory is the "lucky shot" theory: the hypothesis that small arms fire from the crew of the Housatonic may have sufficiently damaged the submarine to sink it. However, ballistic experiments with cast iron samples, analysis of historical experiments firing Civil War-era projectiles at cast iron samples, and calculation of the tidal currents and sinking trajectory of the submarine indicate that this theory is not likely. Based on our results, the "lucky shot" theory does not explain the sinking of the world's first successful combat submarine.


Molecular cloning and characterization of cDNA for eukaryotic transcription factor S-II.
Complementary DNA (cDNA) clones encoding a transcription factor S-II were isolated and characterized. The primary structure of S-II was determined by nucleotide sequence analysis of these clones. The predicted primary structure was consistent with the model that we proposed previously from the results of biochemical analyses of S-II. Using these clones as probes, we analyzed the mRNA for S-II. RNA blot analysis demonstrated the presence of four species of mRNA that hybridized with S-II cDNA in Ehrlich ascites tumor cells. This is the first evidence of polymorphism of mRNA encoding a transcription factor of RNA polymerase II. The results of analysis of the genomic structure suggested that the polymorphism of mRNA may be due to alternative splicing, or differences in initiation or termination of transcription.


Ferrous intraocular foreign bodies and magnetic resonance imaging.
We placed various sized ferrous foreign bodies in and around the eye, sclera, and orbit in a rabbit model before performing magnetic resonance imaging studies. All foreign bodies were detected on plain x-rays. Only one of the largest fragments (3 x 1 x 1 mm) demonstrated movement on exposure to the magnetic field.


Medicare coverage and reimbursement of outpatient prescription drugs in the US: history, recent changes and outlook for the future.
US Medicare provides health insurance to 41.5 million disabled and elderly Americans. Outpatient coverage, including a limited pharmacy benefit, is provided by Medicare Part B. Even with very restrictive criteria, Part B incurs 8.5 billion US dollars annually for outpatient prescription medications. Prior to passage of the Medicare Prescription Drug and Improvement Modernization Act (MMA) 2003, the Part B pharmacy benefit was criticised for its limited coverage criteria and flawed reimbursement practices. Despite the changes made under the MMA, these two issues continue to be concerns for the Medicare Part B outpatient prescription drug programme because (a) the criteria for selection of drugs for coverage do not necessarily reflect valuable advances in medicine, and the extent to which the new private-sector style 2006 Medicare Part D drug benefit will correct this is unknown; and (b) although pre-MMA average wholesale price-based reimbursement practices were clearly flawed, MMA changes such as use of the average sales price and providing increased reimbursement to physicians for drug administration may or may not be successful, and could lead to new problems. The extent to which the MMA and its associated Part D drug benefit address concerns and advance towards better, more cost-effective healthcare is reviewed and recommendations made.


Improving palliative care through teamwork (IMPACTT) in nursing homes: Study design and baseline findings.
The 2014 Institute of Medicine report recommended that healthcare providers caring for individuals with advanced illness have basic palliative care competencies in communication, inter-professional collaboration, and symptom management. Nursing homes, where one in three American decedents live and die, have fallen short of these competency goals. We implemented an intervention study to examine the efficacy of nursing home-based integrated palliative care teams in improving the quality of care processes and outcomes for residents at the end of life. This paper describes the design, rationale, and challenges of a two-arm randomized controlled trial of nursing home-based palliative care teams in 31 facilities. The impact of the intervention on residents' outcomes is measured with four risk-adjusted quality indicators: place of death (nursing home or hospital), number of hospitalizations, and self-reported pain and depression in the last 90-days of life. The effect of the intervention is also evaluated with regard to staff satisfaction and impact on care processes (e.g. palliative care competency, communication, coordination). Both secondary (e.g. the Minimum Data Set) and primary (e.g. staff surveys) data are employed to examine the effect of the intervention. Several challenges in conducting a complex, nursing home-based intervention have been identified. While sustainability of the intervention without research funding is not clear, we surmise that without changes to the payment model that put palliative care services in this care setting on par with the more "skilled" care, it will not be reasonable to expect any widespread efforts to implement facility-based palliative care services.


Efficacy of virtual plastic surgery encounters in establishment of care and surgical conversion.
The efficacy of virtual visits in converting new patients into established patients undergoing surgical treatment has not been demonstrated. The aim of this study was to evaluate patient retention and surgical conversion rate after an initial virtual plastic surgery consultation. An IRB-approved retrospective review of all new plastic surgery patients seen between May and August 2020 at a single institution was conducted. The initial encounter type, chief complaint, demographics, treatment recommendation, insurance approval rate, number and modality of pre- and postoperative visits, time to procedure, follow up, and complications were recorded. Patient retention and surgery conversion rate were calculated. Statistical analysis was performed with Chi-squared test, Fisher's exact test, and unpaired t-test. In total, the records of 1889 new patients were reviewed (1635 in-person, 254 virtual). Virtual patients were younger (44.5 ± 19.0 versus 49.5 ± 20.7 years, p < 0.001), and nearly half resided greater than 50 miles away (42% versus 16%, p < 0.001). Virtual patients more frequently presented for cosmetic surgery (14% versus 7%, p < 0.001), lymphedema (15% versus 3%, p < 0.001), and gender dysphoria (11% versus 2%, p < 0.001). In-person patients presented more often for trauma (18% versus 5%, p < 0.001), elective hand complaints (16% versus 3%, p < 0.001), and breast reconstruction (9% versus 4%, p < 0.01). There were no differences in patient retention (p = 0.45) and procedure conversion rate (p = 0.21) between the groups. Telemedicine provides an opportunity to increase the practice catchment area and is as effective as in-person first encounters for establishing care and transition to surgery.


Transcriptomic survey on the microcystins production and growth of Microcystis aeruginosa under nitrogen starvation.
Cyanobacteria are a vital component of freshwater phytoplankton, and many species are recognized for their ability to produce toxins and harmful algal blooms (HABs). Nitrogen is an essential element of all the complex macromolecules in algal cells. However, the underlying molecular mechanism of the changes in transcriptomic patterns and physiological responses in response to N starvation is poorly understood. The transcriptomes were generated via RNA-sequencing (RNA-Seq) technology to study the major metabolic pathway under N starvation. The results shed light on the mechanism of toxin production and physiological adaptations in Microcystis aeruginosa (M. aeruginosa). The cell density gradually increased during the first two days then declined over time and was finally stable at (15.50 ± 0.5) × 105 cell mL-1 after 6 days. The chlorophyll-a content and phycocyanin content of M. aeruginosa increased during the first two days and subsequently decreased markedly over time under N starvation. The variable to maximum chlorophyll fluorescence ratio (Fv/Fm ratio) decreased with time under N starvation. Most photosynthesis genes have similarity decreasing trends with growth physiological changes. The microcystins (MCs) levels generally increased first, reaching a peak value with 1.35 pg cell-1 on the fifth day, and then remained roughly constant. The genes involved in N metabolism-related gene expression were upregulated to maintain normal biological activity, while the genes involved in photosynthesis-related gene expression were downregulated to save energy. All genes encoding algae toxin synthesis were upregulated under N starvation. The observed expression patterns demonstrate that all MCs genes respond similarly to MCs production within the cell. Our results indicate the response mechanism of M. aeruginosa under N starvation and provide a comprehensive understanding of N-controlling cyanobacteria and MCs synthesis.


The 2-5A system in viral infection and apoptosis.
The 2-5A system is an established endogenous antiviral pathway. Interferon treatment of cells leads to an increase in basal, but latent, levels of 2-5A-dependent RNase (RNase L) and the family of 2'-5' oligoadenylate synthetases (OAS). Double-stranded RNA, thought to be derived from viral replication intermediates, activates OAS. Activated OAS converts ATP into unusual short 2'-5' linked oligoadenylates called 2-5A [ppp5'(A2'p5')2A]. The 2-5A binds to and activates RNase L which cleaves single stranded RNA with moderate specificity for sites 3' of UpUp and UpAp sequences, and thus leads to degradation of cellular rRNA. During apoptosis, generalized cellular RNA degradation, distinct from the differential expression of mRNA species that may regulate specific gene expression during apoptosis, has been observed. The mechanism of RNA breakdown during apoptosis has been commonly considered a non-specific event that reflects the generalized shut down of translation and homeostatic regulation during cell death. Due to the similar RNA degradation that occurs during both apoptosis and viral infection we investigated the potential role of RNase L in apoptosis. To investigate whether RNase L activity could lead to apoptosis, NIH3T3 cells were transfected with a lac-inducible vector containing the human RNase L gene. Treatment of these cells with isopropylthiogalactoside (IPTG) caused loss of cell viability that was confirmed as an apoptotic cell death by morphological and biochemical criteria. Similarly, specific allosteric activation of endogenous RNase L by introduction of 2-5A directly into L929 cells also induced apoptosis. In L929 cells poly(I).poly(C) treatment in combination with interferon caused an increase in apoptosis whereas neither interferon or double stranded RNA alone altered cell viability. Therefore, increased expression or activation of RNase L causes apoptosis. Inhibition of RNase L, specifically with a dominant negative mutant, suppressed poly(I)Ypoly(C)-induced apoptosis in interferon-primed fibroblasts. Poliovirus, a picornovirus with a single-stranded RNA genome, causes apoptosis of HeLa cells. Expression of the dominant negative inhibitor of RNase L in HeLa prevented virus-induced apoptosis and maintained cell viability. Thus, reduction or inhibition of RNase L activity prevents apoptosis. Both apoptosis and the 2-5A system can provide defense against viral infection in multicellular organisms by preventing production and therefore spread of progeny virus. RNase L appears to function in both mechanisms, therefore, initiation of apoptosis may be one mechanism for the antiviral activity of the 2-5A system.


Hemimyeloschisis associated with hydrocephalus.
Hemimyeloschisis is a very rare congenital malformation of the spine which is characterized by a split cord malformation associated with complete exposureof the one hemicord to the exterior. We report a case of hemimyeloschisis on the right hemicord in a patient with hydrocephalus. The patient was a female neonate with myeloschisis, skin defect, and hypertrichosis visible at birth in the upper lumbar region. There was also skin defect around the myeloschisis. Computed tomography (CT) revealed a bony septum at the L2 level as well as the myeloschisis on the right hemicord, which enabled us to make a precise preoperative diagnosis of this rare malformation. There was also mild hydrocephalus in cranial CT scan. Closure of the myeloschisis and removal of the septum were successfully performed associated with the closure of the skin in one stage surgery to prevent subsequent infection and neurological deterioration. Ventriculoperitoneal shunt insertion was performed for hydrocephalus. The presence of combined split cord malformation and myeloschisis is consistent with the hypothesis of an ontogenic basis of development. This combination also emphasizes the importance of early imaging for diagnosis in this complex malformation.


Are hospitalized or ambulatory patients with heart failure treated in accordance with European Society of Cardiology guidelines? Evidence from 12,440 patients of the ESC Heart Failure Long-Term Registry.
To evaluate how recommendations of European guidelines regarding pharmacological and non-pharmacological treatments for heart failure (HF) are adopted in clinical practice. The ESC-HF Long-Term Registry is a prospective, observational study conducted in 211 Cardiology Centres of 21 European and Mediterranean countries, members of the European Society of Cardiology (ESC). From May 2011 to April 2013, a total of 12,440 patients were enrolled, 40.5% with acute HF and 59.5% with chronic HF. Intravenous treatments for acute HF were heterogeneously administered, irrespective of guideline recommendations. In chronic HF, with reduced EF, renin-angiotensin system (RAS) blockers, beta-blockers, and mineralocorticoid antagonists (MRAs) were used in 92.2, 92.7, and 67.0% of patients, respectively. When reasons for non-adherence were considered, the real rate of undertreatment accounted for 3.2, 2.3, and 5.4% of the cases, respectively. About 30% of patients received the target dosage of these drugs, but a documented reason for not achieving the target dosage was reported in almost two-thirds of them. The more relevant reasons for non-implantation of a device, when clinically indicated, were related to doctor uncertainties on the indication, patient refusal, or logistical/cost issues. This pan-European registry shows that, while in patients with acute HF, a large heterogeneity of treatments exists, drug treatment of chronic HF can be considered largely adherent to recommendations of current guidelines, when the reasons for non-adherence are taken into account. Observations regarding the real possibility to adhere fully to current guidelines in daily clinical practice should be seriously considered when clinical practice guidelines have to be written.


The Use of a Low-Fidelity Simulator to Improve Vascular Anastomosis Skills of Residents during the COVID-19 Pandemic.
To evaluate a workshop using a low-fidelity simulator for training vascular surgery residents in vascular anastomosis during the COVID-19 pandemic. Prospective, controlled, single-center. Vascular surgery residents at the São Paulo University Medical School were enrolled in the COVID Group (five post graduation year 3 residents) or Control Group (five PGY-4 residents). The COVID Group was trained via a vascular anastomosis workshop. The residents were evaluated using Objective Structured Assessment of Technical Skills (OSATS), Final Product Analysis and time to perform the procedure. The number of anastomoses performed by the residents were calculated. Data were subjected to statistical analysis, and P < 0.05 was considered significant. There was a significant reduction in the number of vascular anastomoses performed by the residents between the COVID group and the control group (mean 22.6 ± 7.76 vs. 35.2 ± 3.9, P = 0.01, Student's t-test). Before the workshop, 80% of the residents from the COVID group failed to perform a vascular anastomosis on the simulator. During the workshop, there was improvement in the Objective Structured Assessment of Technical Skills (OSATS) score (initial: 16.5, interquartile range (IQR) 0, under supervision: 25, IQR 5, and at the end of the workshop: 26.5, IQR 2.5; P = 0.049, Friedman's test) and in the Final Product Analysis (initial: 14.5, IQR 6, under supervision: 26.5, IQR 4.625, end of the workshop: 27, IQR 4, P = 0.049, Friedman's test). Time was not significantly different (initial: 35.6, IQR 2.77; under supervision: 25.8 min, IQR 4.53; P = 0.07, Friedman's test). The residents' technical scores were stable 6 months after the training, and there was no difference between their final scores and those of the control group. The residents from the COVID Group reported an improvement in their knowledge, technical skills and confidence after the workshop. A workshop using a low-fidelity simulator improved vascular surgery residents' skills and confidence in vascular anastomosis during the pandemic year, when they performed fewer surgical procedures.


Easy access to α-ketoamides as SARS-CoV-2 and MERS Mpro inhibitors via the PADAM oxidation route.
SARS-CoV-2 caused worldwide the current outbreak called COVID-19. Despite multiple countermeasures implemented, there is an urgent global need for new potent and efficient antiviral drugs against this pathogen. In this context, the main protease (Mpro) of SARS-CoV-2 is an essential viral enzyme and plays a pivotal role in viral replication and transcription. Its specific cleavage of polypeptides after a glutamine residue has been considered as a key element to design novel antiviral drugs. Herein, we reported the design, synthesis and structure-activity relationships of novel α-ketoamides as covalent reversible inhibitors of Mpro, exploiting the PADAM oxidation route. The reported compounds showed μM to nM activities in enzymatic and in the antiviral cell-based assays against SARS-CoV-2 Mpro. In order to assess inhibitors' binding mode, two co-crystal structures of SARS-CoV-2 Mpro in complex with our inhibitors were solved, which confirmed the covalent binding of the keto amide moiety to the catalytic Cys145 residue of Mpro. Finally, in order to interrogate potential broad-spectrum properties, we assessed a selection of compounds against MERS Mpro where they showed nM inhibitory potency, thus highlighting their potential as broad-spectrum coronavirus inhibitors.


Interventions Employed By Licensed Nurses in Nursing Homes: Refinement and Validation of an Existing Omaha System Nursing Intervention Set.
Background and Purpose: Demands on long-term services and supports for older adults are growing, although geriatric workforce shortages have persisted for decades. Methods to define and quantify practice of licensed nurses in nursing homes are needed for work optimization within limited nurse resources available in nursing homes. This study aimed to refine and validate observable nursing interventions for nursing homes, using the Omaha System. Methods: Based on the existing corpus of Omaha System interventions for acute care nursing, this multi-phase, multi-method study included a mapping procedure of interviews from licensed nurses in nursing homes, the evaluation of content validity and coding of the interventions using a survey, and inter-observer reliability assessment using TimeCaT. Results: This study validated 57 observable interventions for nursing homes. Of the previously identified acute care nursing interventions, eight interventions were deemed out of scope. One additional intervention was identified. Refined intervention definitions were related to procedures common in acute care settings such as tracheal intubations/extubations and nasogastric tube insertion that were not performed in nursing homes. Expert agreement for content validity and coding of the interventions was high (S-CVI = 0.97), and inter-observer reliability levels (Cohen's κ value >0.4; proportion agreement >60%) were acceptable for all case studies. Implications for Practice: The validated observable Omaha System nursing interventions for nursing home practice have potential for use in future studies of nursing home practice to understand evidence-based practice, and gaps in care provided. The methodology may be extended to define observable interventions for other roles and settings.


Late recurrence of breast cancer is associated with pro-cancerous immune microenvironment in the primary tumor.
The fact that 20-40% of all breast cancer (BC) patients develop recurrence when 5 year survival is 90% strongly suggests that late recurrence, i.e. more than 5 years after diagnosis, is the remaining challenge to decrease the absolute number of BC deaths. Better understanding late recurrence is an essential first step to address this issue. We hypothesized that primary tumors with a distinctive tumor immune microenvironment will develop late recurrence. Accordingly, we evaluated the relationship between the timing of cancer recurrence, clinical factors, gene expression profiles, and immune status utilizing two published large cohorts. 308 primary BCs in TCGA were analyzed and categorized as: recurrence ≤2 years (Early, n = 49), between 2-5 years (Mid, n = 54), recurrence >5 years (Late, n = 20), and no recurrence >5 years (Survivors, n = 185). 1,727 primary BCs in METABRIC were analyzed and categorized similarly: Early, n = 170; distant (D), n = 19; local (L), Mid, n = 213; D, n = 21; L, Late, n = 199; D, n = 57, L, and Survivors, n = 1048. Utilizing pre-ranked GSEA, we showed that primary tumors with Survivors were associated with anti-cancer signaling such as INF-α/-γ response and TNF-α signaling, compared with all recurrence groups in pre-ranked GSEA. Furtherrmore, we found that host defense immunity (leukocyte fraction, lymphocyte infiltration, and macrophage fractions) was decreased in primary tumors with Late recurrence compared with Survivors. Utilizing the CIBERSORT algorithm, we showed anti-cancer lymphocytes, memory CD4+ T cells and γδT cells, were significantly lower, and pro-cancerous regulatory T cells were significantly higher in Late tumors compared with Survivors. In agreement, cytolytic activity score that assesses immune cell cytolytic activity was significantly lower in Late compared with Survivors. We demonstrated that not only host defense immunity, but also pro-cancerous immune cells and immune cell cytolytic activity in primary BC was associated with late recurrence.


Comparison of acute central nervous system and cardiovascular toxicity of 2-chloroprocaine and prilocaine in the rat.
In this study, we compared the central nervous system and cardiovascular system toxicity of i.v. administered 0.5% 2-chloroprocaine (N = 10) and 0.5% prilocaine (N = 10) in lightly anaesthetised rats. Arterial blood pressure, ECG and EEG were continuously recorded. Prilocaine produced the predetermined toxic end-points (i.e. seizure activity on EEG, isoelectric EEG, cardiac arrhythmia on ECG, asystole on ECG) at significantly lower doses than 2-chloroprocaine (P < 0.05). The mean dose of prilocaine producing asystole was 166 mg.kg-1 (+/- 45 mg.kg-1, s.d.) vs. 255 mg.kg-1 (+/- 42 mg.kg-1) for 2-chloroprocaine (P < 0.01). The rate of decrease of mean arterial blood pressure during the infusion was significantly faster with prilocaine (P < 0.01). Typically, arrhythmias did not appear until just before asystole, suggesting that neither of the local anaesthetics possessed marker arrhythmogenic properties. It is concluded that prilocaine is slightly more toxic than 2-chloroprocaine in the rat, but that both local anaesthetics have a wide margin of safety. Doses producing seizure activity on the EEG (prilocaine 53 mg.kg-1 and 2-chloroprocaine 70 mg.kg-1, on average) are much higher than those used in clinical practice (usually < 10 mg.kg-1).


Probit analysis applied to the allergen dose-response curve: a method for epidemiologic surveys.
The degree of skin sensitivity to five common allergens (grass, Dermatophagoides pteronyssinus, mugwort, birch, and Parietaria) was determined by the threshold dilution technique in all the skin test reactors of a random sample of 295 schoolchildren (142 male and 153 female subjects, age range 11 to 14 years), and the frequency distribution of responders at each concentration was analyzed by probit analysis. The potency of each allergen was presented in terms of median effective dose (ED50), and comparison between different allergens and between symptomatic and asymptomatic subjects was made by computing the relative potency. The ED50 of the allergens was found to be nearly identical (grass, 13.5; D. pteronyssinus, 12.3; mugwort, 9.6 activity units by RAST (AUR)/ml) with the exception of birch (22 AUR/ml). Grass and D. pteronyssinus demonstrated a lower ED50 in symptomatic subjects (3.3 and 2.8 AUR/ml, respectively) than in asymptomatic subjects (85 and 27 AUR/ml, respectively). The lower fiducial limits of ED50 in symptomatic subjects demonstrated to be a cutoff point, since they included only 5% and 12% of asymptomatic reactors to grass and D. pteronyssinus, respectively. We conclude that probit analysis applied to the distribution of threshold doses of allergen extracts is a useful method to evaluate skin sensitivity in epidemiologic surveys. We believe that the ED50 is a practical and reliable allergy index.


Dietary restriction improves repopulation but impairs lymphoid differentiation capacity of hematopoietic stem cells in early aging.
Dietary restriction (DR) improves health, delays tissue aging, and elongates survival in flies and worms. However, studies on laboratory mice and nonhuman primates revealed ambiguous effects of DR on lifespan despite improvements in health parameters. In this study, we analyzed consequences of adult-onset DR (24 h to 1 yr) on hematopoietic stem cell (HSC) function. DR ameliorated HSC aging phenotypes, such as the increase in number of HSCs and the skewing toward myeloid-biased HSCs during aging. Furthermore, DR increased HSC quiescence and improved the maintenance of the repopulation capacity of HSCs during aging. In contrast to these beneficial effects, DR strongly impaired HSC differentiation into lymphoid lineages and particularly inhibited the proliferation of lymphoid progenitors, resulting in decreased production of peripheral B lymphocytes and impaired immune function. The study shows that DR-dependent suppression of growth factors and interleukins mediates these divergent effects caused by DR. Supplementation of insulin-like growth factor 1 partially reverted the DR-induced quiescence of HSCs, whereas IL-6/IL-7 substitutions rescued the impairment of B lymphopoiesis exposed to DR. Together, these findings delineate positive and negative effects of long-term DR on HSC functionality involving distinct stress and growth signaling pathways.


Dinuclear ruthenium(II) polypyridyl complexes containing large, redox-active, aromatic bridging ligands: synthesis, characterization, and intramolecular quenching of MLCT excited states.
Two new ruthenium(II) polypyridyl dimers containing the large planar aromatic bridging ligands 9,11,20,22-tetraazatetrapyrido[3,2-a:2'3'-c:3' ',2''-l:2''',3'''-n]pentacene (tatpp) and 9,11,20,22-tetraazatetrapyrido[3,2-a:2'3'-c:3'',2' '-l:2''',3'''-n]pentacene-10,21-quinone (tatpq) have been synthesized and characterized by (1)H and (13)C NMR, MALDI mass spectrometry, and elemental analyses. The electronic properties (UV-vis, redox, photophysical) of these dimers are analyzed in the context of orbital calculations (PM3 level) on the bridging ligands. A localized orbital model is proposed in which low-lying acceptor orbitals on the center portion of the ligands effectively quench the Ru(II)-based MLCT emission via a mechanism that can be viewed as intramolecular electron transfer to specific subunits of the bridges.


Application of Laser Capture Microdissection to Craniofacial Biology: Characterization of Anatomically Relevant Gene Expression in Normal and Craniosynostotic Rabbit Sutures.
Fusion of the cranial sutures is thought to depend on signaling among perisutural tissues. Mapping regional variations in gene expression would improve current models of craniosynostosis. Laser capture microdissection (LCM) isolates discrete cell populations for gene expression analysis. LCM has rarely been used in the study of mineralized tissue. This study sought to evaluate the potential use of LCM for mapping of regional gene expression within the cranial suture. Coronal sutures were isolated from 10-day-old wild-type and craniosynostotic (CS) New Zealand White rabbits, and LCM was used to isolate RNA from the sutural ligament (SL), osteogenic fronts (OF), dura mater, and periosteum. Relative expression levels for Fibroblast Growth Factor 2 (FGF2), Fibroblast Growth Factor Receptor 2 (FGFR2), Transforming Growth Factor Beta 2 (TGFβ-2), Transforming Growth Factor Beta 3 (TGFβ-3), Bone Morphogenetic Protein 2 (BMP-2), Bone Morphogenetic Protein 4 (BMP-4), and Noggin were determined using quantitative real-time PCR. A fivefold increase in TGFβ2 expression was detected in the CS SL relative to wild type, whereas 152-fold less TGFβ-3 was detected within the OF of CS animals. Noggin expression was increased by 10-fold within the CS SL, but reduced by 13-fold within the CS dura. Reduced expression of FGF2 was observed within the CS SL and dura, whereas increased expression of FGFR2 was observed within the CS SL. Reduced expression of BMP-2 was observed in the CS periosteum, and elevated expression of BMP-4 was observed in the CS SL and dura. LCM provides an effective tool for measuring regional variations in cranial suture gene expression. More precise measurements of regional gene expression with LCM may facilitate efforts to correlate gene expression with suture morphogenesis and pathophysiology.


The impact of total joint replacement in rheumatoid arthritis.
Management of the patient with rheumatoid arthritis (RA) requires a multidisciplinary approach, the role of the surgeon being to improve functional ability for the patient by reconstructing a deteriorated joint by total joint arthroplasty (TJA). An advantage of prosthetic evaluation over pharmacological medication evaluations is that the 'compliance' of the patient with the treatment (i.e. the TJA) is 100%, even at long-term follow-up. However, long-term follow-up of prosthesis evaluation is as difficult as the evaluation of any other intervention. Although the goal of any intervention on an RA patient is to improve functional ability, and thus self-support, of the patient, objective evaluation of the surgical procedure, and of its impact on the patient, can be difficult. The potential chronic course of RA makes evaluation of a specific surgical procedure and its effect on the patient difficult to interpret. The success of the TJA is generally judged on a survivorship analysis at 10 or 15 years in national registries (i.e. >40000 implants); revision surgery is used as an end-point for survival of the TJA. With a mean 90% survival at 10-year follow-up, total hip arthroplasty and total knee arthroplasty may be considered gold-standard TJA procedures for the patient. While revision is the end-point, the course to this end-point starts with progressive micromotion of the prosthesis. The effect of prosthetic changes, and of medication on prosthesis migration, can be measured very accurately by radiostereometry. The latter measures the actual performance of the TJA in the bone. Next to these more procedure-oriented evaluations, patient-oriented evaluations (e.g. quality of life, patient expectations) are of importance in judging the impact of the TJA on the RA patient. These evaluations provide evidence that the pre-operative status of the joint/extremity determines the extent of post-operative functional gain. Thus, postponing TJA for too long will give less functional benefit.


Polymorphisms in the long non-coding RNA CDKN2B-AS1 may contribute to higher systolic blood pressure levels in hypertensive patients.
Hypertension (HT) is a complex disorder influenced by both genetic and environmental factors. Recent genome-wide association studies have identified a major risk locus for atherosclerosis on chromosome 9p21.3. SNPs within the coding sequences of CDKN2A/B and the long non-coding RNA CDKN2B-AS1 could potentially contribute to HT development. Thus, this study aimed to investigate whether the frequency of four SNPs on chromosome 9p21.3 affects blood pressure (BP) levels in Turkish HT patients, and to examine correlations between these SNPs, specific SNP haplotypes, and HT. This is a case-control study comparing HT patients and healthy controls. Real-time polymerase chain reaction (RT-PCR) analysis was utilized to detect SNPs rs10757274, rs2383207, rs10757278, and rs1333049 in 170 HT patients and 180 healthy controls. Each SNP was detected at significantly higher frequencies in HT patients than in controls (p values 0.001); however, there was no significant link between rs10757274, rs2383207, rs10757278, and rs1333049 SNPs and HT grades. Furthermore, there was a significant association between elevated systolic BP levels and rs1333049 GG genotype (p=0.047), while weight gain and increased fasting glucose levels were significantly associated with rs2383207 AA genotype (p=0.020 and p=0.009, respectively). Lastly, we detected a correlation between GG, GA, and AG haplotypes in block 1 (rs10757274, rs2383207) and GC and AG haplotypes in block 2 (rs10757278, rs1333049) and HT. Our findings suggest that SNPs rs10757274, rs2383207, rs10757278, and rs1333049, particularly those within the CDKN2B-AS1 gene, and related haplotypes may confer increased susceptibility to HT development.


Fungi associated with black mould on baobab trees in southern Africa.
There have been numerous reports in the scientific and popular literature suggesting that African baobab (Adansonia digitata) trees are dying, with symptoms including a black mould on their bark. The aim of this study was to determine the identity of the fungi causing this black mould and to consider whether they might be affecting the health of trees. The fungi were identified by sequencing directly from mycelium on the infected tissue as well as from cultures on agar. Sequence data for the ITS region of the rDNA resulted in the identification of four fungi including Aureobasidium pullulans, Toxicocladosporium irritans and a new species of Rachicladosporium described here as Rachicladosporium africanum. A single isolate of an unknown Cladosporium sp. was also found. These fungi, referred to here as black mould, are not true sooty mould fungi and they were shown to penetrate below the bark of infected tissue, causing a distinct host reaction. Although infections can lead to dieback of small twigs on severely infected branches, the mould was not found to kill trees.


Complete Genome Sequence of the Deep-Sea Bacterium Psychromonas Strain CNPT3.
Members of the genus Psychromonas are commonly found in polar and deep-sea environments. Here we present the genome of Psychromonas strain CNPT3. Historically, it was the first bacterium shown to piezoregulate the composition of its membrane lipids and to have a higher growth rate at 57 megapascals (MPa) than at 0.1 MPa.


Comparison of T-Spot.TB and tuberculin skin test among silicotic patients.
In the present study, T-Spot.TB and the tuberculin skin test (TST) were compared in the screening of latent tuberculosis infection among silicotic patients. A conditional probability model was used to compare the potential clinical utilities of T-Spot.TB and TST performed on 134 silicotic subjects from December 1, 2004 to January 31, 2007. Data from a historical cohort were also reanalysed for further comparison. Agreement with T-Spot.TB was best using a TST cut-off of 10 mm. Age >or=65 yrs independently predicted a tuberculin reaction <10 mm (odds ratio = 3), but not a negative T-Spot.TB response. Lower measures of agreement were observed among current smokers and those aged >or=65 yrs. Tuberculin reaction size was well correlated with both early secretary antigenic target 6 and culture filtrate protein 10 spot counts, except among current smokers. Within the current estimates of sensitivity (88-95%) and specificity (86-99%) for T-Spot.TB, the positive likelihood ratio for T-Spot.TB test would be substantially higher (6.29-95.0 versus 1.65-1.94) and negative likelihood ratio substantially lower (0.05-0.14 versus 0.32-0.41) than the corresponding ratios for the tuberculin test. A low tuberculosis risk differential was similarly observed between tuberculin-negative and untreated tuberculin-positive subjects in the historical cohort. T-Spot.TB is likely to perform better than tuberculin test in the screening of latent tuberculosis infection among silicotic subjects.


Phylogenetic studies of complete mitochondrial DNA molecules place cartilaginous fishes within the tree of bony fishes.
It is commonly acknowledged that cartilaginous fishes, Chondrichthyes, have a basal position among the Gnathostomata (jawed vertebrates). In order to explore this relationship we have sequenced the complete mitochondrial genome of the spiny dogfish, Squalus acanthias, and included it in a phylogenetic analysis together with a number of bony fishes and amniotes. The phylogenetic reconstructions placed the dogfish among the bony fishes. Thus, and contrary to the common view, the analyses have shown that the position of the sharks is not basal among the gnathostomes. The presently recognized phylogenetic position of the dogfish was identified irrespective of the outgroup used, echinoderms or agnathan fishes. The lungfish was the most basal gnathostome fish, while the teleosteans had an apical position in the piscine tree. A basal position of the dogfish among the gnathostomes was statistically rejected, but the phylogenetic relationship among the coelacanth, spiny dogfish, and teleosts was not conclusively resolved. The findings challenge the current theory that sharks and other chondrichthyans, if monophyletic, are the sister group to all other extant gnathostomes. The results open to question the status of several morphological characters commonly used in piscine phylogenetic reconstruction, most notably the presence versus absence of endochondral bone in the endoskeleton, the macromeric versus micromeric structure of the exoskeleton, and the presence/absence of swimbladder and/or lung. The study also confirmed recent findings demonstrating that the origin of the amniotes is deeper than the diversification of extant bony fishes.


Three patients with autoimmune cholangiopathy treated with prednisolone.
We describe three patients with autoimmune cholangiopathy, i.e., anti-mitochondrial antibody-negative and anti-nuclear antibody-positive primary biliary cirrhosis, who were treated with prednisolone. Serum anti-mitochondrial antibody and anti-pyruvate dehydrogenase-E2 component antibody were determined by immunofluorescence of frozen sections and enzyme-linked immunosorbent assay, respectively. Immunoblotting using mitochondria prepared from rat liver was performed to analyze anti-mitochondrial antibody in detail. Serum from one patient reacted with a 48-kilodalton protein, but sera from the other two patients failed to react with the mitochondrial proteins. There was a marked improvement in liver function test results after prednisolone treatment. Before treatment, liver biopsy in all three patients showed histological features of primary biliary cirrhosis with hepatocellular necrosis. Repeat biopsy during treatment showed marked amelioration of hepatocellular damage in all three patients, although bile duct involvement persisted in two patients. These findings suggest that prednisolone is an effective treatment for hepatocellular damage in patients with autoimmune cholangiopathy, but has little impact on the bile duct involvement.


Human torovirus: a new virus associated with neonatal necrotizing enterocolitis.
Toroviruses have been associated with gastroenteritis in both animals and humans. The aim of this study was to examine the fecal excretion of torovirus in infants with necrotizing enterocolitis (NEC). We reviewed all infants with NEC admitted to our tertiary care NICU over a 5-y period who had stool specimens sent for microbial culture and virology. Infants in the NICU during the same period with diagnoses other than NEC served as controls. Forty-four infants with NEC stages I-III were identified, and pathogenic organisms were identified in 27 (61%). Toroviruses were identified in stool cultures in 48% of patients with NEC, and 17% of the non-NEC controls (p<0.001). There was no significant difference in illness severity or mortality between the torovirus-positive and -negative infants with NEC. Torovirus should be added to the list of infectious agents associated with NEC in newborn infants. The exact role torovirus plays in the etiology and progression of NEC warrants further investigation.