Human skeletal muscle sympathetic nerve activity, heart rate and limb haemodynamics with reduced blood oxygenation and exercise.
Acute systemic hypoxia causes significant increases in human skeletal muscle sympathetic nerve activity (MSNA), heart rate and ventilation. This phenomenon is thought to be primarily mediated by excitation of peripheral chemoreceptors sensing a fall in arterial free oxygen partial pressure (Pa,O2). We directly tested the role of Pa,O2 on MSNA (peroneal microneurography), heart rate, ventilation and leg haemodynamics (n = 7-8) at rest and during rhythmic handgrip exercise by using carbon monoxide (CO) to mimic the effect of systemic hypoxia on arterial oxyhaemoglobin (approximately 20 % lower O2Hba), while normalising or increasing Pa,O2 (range 40-620 mmHg). The four experimental conditions were: (1) normoxia (Pa,O2 approximately 110 mmHg; carboxyhaemoglobin (COHb) approximately 2 %); (2) hypoxia (Pa,O2 approximately 40 mmHg; COHb approximately 2 %); (3) CO + normoxia (Pa,O2 approximately 110 mmHg; COHb approximately 23 %); and (4) CO + hyperoxia (Pa,O2 approximately 620 mmHg; COHb ~24 %). Acute hypoxia augmented sympathetic burst frequency, integrated MSNA, heart rate and ventilation compared to normoxia over the entire protocol (7-13 bursts min-1, 100-118 %, 13-17 beats min-1, 2-4 l min-1, respectively, P < 0.05). The major new findings were: (1) CO + normoxia and CO + hyperoxia also elevated MSNA compared to normoxia (63-144 % increase in integrated MSNA; P < 0.05) but they did not increase heart rate (62-67 beats min-1) or ventilation (6.5-6.8 l min-1), and (2) despite the 4-fold elevation in MSNA with hypoxaemia and exercise, resting leg blood flow, vascular conductance and O2 uptake remained unchanged. In conclusion, the present results suggest that increases in MSNA with CO are not mediated by activation of the chemoreflex, whereas hypoxia-induced tachycardia and hyperventilation are mediated by activation of the chemoreflex in response to the decline in Pa,O2. Our findings also suggest that Pa,O2 is not an obligatory signal involved in the enhanced MSNA with reduced blood oxygenation.


Coronin-1C Protein and Caveolin Protein Provide Constitutive and Inducible Mechanisms of Rac1 Protein Trafficking.
Sustained directional fibroblast migration requires both polarized activation of the protrusive signal, Rac1, and redistribution of inactive Rac1 from the rear of the cell so that it can be redistributed or degraded. In this work, we determine how alternative endocytic mechanisms dictate the fate of Rac1 in response to the extracellular matrix environment. We discover that both coronin-1C and caveolin retrieve Rac1 from similar locations at the rear and sides of the cell. We find that coronin-1C-mediated extraction, which is responsible for Rac1 recycling, is a constitutive process that maintains Rac1 protein levels within the cell. In the absence of coronin-1C, the effect of caveolin-mediated endocytosis, which targets Rac1 for proteasomal degradation, becomes apparent. Unlike constitutive coronin-1C-mediated trafficking, caveolin-mediated Rac1 endocytosis is induced by engagement of the fibronectin receptor syndecan-4. Such an inducible endocytic/degradation mechanism would predict that, in the presence of fibronectin, caveolin defines regions of the cell that are resistant to Rac1 activation but, in the absence of fibronectin leaves more of the membrane susceptible to Rac1 activation and protrusion. Indeed, we demonstrate that fibronectin-stimulated activation of Rac1 is accelerated in the absence of caveolin and that, when caveolin is knocked down, polarization of active Rac1 is lost in FRET experiments and culminates in shunting migration in a fibrous fibronectin matrix. Although the concept of polarized Rac1 activity in response to chemoattractants has always been apparent, our understanding of the balance between recycling and degradation explains how polarity can be maintained when the chemotactic gradient has faded.


No evidence for cancer-related CD44 splice variants in primary and metastatic colorectal cancer.
The expression of alternatively spliced CD44 adhesion molecules has been implicated in the pathogenesis and metastasis of colorectal cancer. Using a new set of primers for exon-specific reverse transcription-polymerase chain reaction (RT-PCR) we delineated the exact exon composition of CD44 mRNAs in normal colorectal mucosa, including isolated colonic crypts, in colorectal carcinomas and in their hepatic metastases. In addition, the surface expression of CD44 isoforms was analysed by immunohistochemistry. We identified by RT-PCR eight variant transcripts expressed in colorectal carcinomas and their metastases, but also constitutively in normal colorectal epithelia. In the normal colorectal epithelium, the surface expression of CD44 standard and variant molecules was restricted to proliferating cells at the bottom of the crypts. Despite expression of these transcripts in colorectal cancers and their metastases, monoclonal antibodies specific for standard or variant epitopes encoded by exons v5 and v6 stained only a few neoplastic lesions. These data point to a differentiation-specific CD44 expression and splicing pattern in proliferating colorectal epithelia. However, they do not support a cancer- or metastasis-specific CD44 splicing pattern. Instead, cell surface availability of CD44 epitopes was reduced rather than increased in primary tumours and particularly in liver metastases.


A direct inhibitory action of prostaglandins upon ACTH secretion at the late stages of the secretory pathway of AtT-20 cells.
1. The mouse AtT-20/D16-16 anterior pituitary tumour cell line was used as a model system for the study of the effects of prostaglandins upon the late stages of the adrenocorticotrophin (ACTH) secretory pathway. 2. Calcium (1 nM - 100 microM), guanosine-5'-O-(3-thiotriphosphate) (GTP-gamma-S) (1 - 100 microM) and mastoparan (1 and 10 microM) all stimulated ACTH secretion from permeabilized AtT-20 cells in a concentration-dependent manner. GTP-gamma-S and mastoparan stimulated ACTH secretion from permeabilized cells in the absence of calcium. Co-incubation with prostaglandins E(1) and E(2) (PGE(1), PGE(2)) (10 microM) but not prostaglandin F(2 alpha) (PGF(2 alpha)) (10 microM) significantly inhibited calcium-, GTP-gamma-S and mastoparan-evoked secretion by 30 - 50%. 3. The effects of PGE(1) and PGE(2) upon GTP-gamma-S (100 microM)-, calcium (10 microM)- and mastoparan (10 microM)-evoked secretion were concentration-dependent. PGE(1) significantly inhibited GTP-gamma-S- and calcium-evoked secretion at concentrations of PGE(1) above 1 microM but mastoparan-evoked secretion only at the highest concentration of PGE(1) investigated (10 microM). PGE(2) was much more potent than PGE(1) and significantly inhibited GTP-gamma-S- and calcium-evoked secretion at 10 nM and above and mastoparan-evoked secretion above 1 microM. 4. The inhibitory effects of PGE(1) and PGE(2) upon calcium-, GTP-gamma-S- and mastoparan-stimulated ACTH secretion from permeabilized cells were pertussis toxin (PTX) sensitive. 5. In intact cells PGE(1), PGE(2) and PGF(2 alpha) (1 nM - 10 microM) acting singly had little or no effect upon ACTH secretion. However, only PGE(2) (1 nM - 10 microM) significantly inhibited corticotrophin-releasing factor-41 (CRF-41) (100 nM)-evoked secretion in a concentration dependent manner. 6. The present study finds that prostaglandins of the E series exert an inhibitory action, via a pertussis toxin-sensitive GTP-binding (G)-protein, in the late stages of the ACTH secretory pathway distal to the G-exocytosis (Ge)/calcium point of control.


Programmed death-ligand 1 triggers PASMCs pyroptosis and pulmonary vascular fibrosis in pulmonary hypertension.
Pyroptosis is a pro-inflammatory form of programmed cell death, whose genesis directly depended on caspase-1 activation. Pulmonary hypertension (PH) is a disease characterized, in part, by vascular fibrosis. Up to now, there is no report on the relationship between pyroptosis and vascular fibrosis in PH. Here, we confirmed that pyroptosis had occurred in the media of pulmonary arteries in two PH rat models and hypoxic human pulmonary arterial smooth muscle cells (hPASMCs). Caspase-1 inhibition attenuated the pathogenesis of PH, as assessed by vascular remodeling, right ventricular systolic pressure, right ventricle hypertrophy and hemodynamic parameters of pulmonary vasculature. Moreover, caspase-1 inhibition suppressed pulmonary vascular fibrosis as demonstrated by Masson staining, as well as immunohistochemistry and Western blot analysis of fibrillar collagen. In addition, Programmed death-ligand 1 (PD-L1) was markedly increased in PH, which was regulated by the transcription factor STAT1. Furthermore, PD-L1 knockdown in hPASMCs repressed the onset of hypoxia-induced pyroptosis and fibrosis. Overall, these data identify a critical STAT1-dependent posttranscriptional modification that promotes PD-L1 expression in the pyroptosis of PASMCs to modulate pulmonary vascular fibrosis and accelerate the progression of PH.


Peroneal Tendon Pathology: Treatment and Reconstruction of Peroneal Tears and Instability.
Peroneal tendon pathology is becoming an increasingly recognized source of lateral-sided ankle pain. High clinical suspicion, along with judicious physical examination coupled with confirmatory advanced imaging modalities, are necessary to make an accurate diagnosis and aid in guiding treatment. Peroneal pathology encompasses several distinct conditions. Peroneal tendon tears and injuries to the peroneal retinaculum must be identified to guide treatment. Patients with peroneal pathology report high levels of satisfaction after surgical management with most returning to their preinjury level of function. An early and accurate diagnosis, along with treatment tailored to the individual, is necessary to obtain optimal outcomes.


Multiple keratoacanthomas after megavoltage radiation therapy.
Multiple keratoacanthomas developed on the face of a woman after radiation therapy to the nasal sinuses. Treatment with isotretinoin appeared to shorten her recovery period.


Quality Measurement and Pay for Performance.
Recent debate has focused on which quality measures are appropriate for surgical oncology and how they should be implemented and incentivized. Current quality measures focus primarily on process measures (use of adjuvant therapy, pathology reporting) and patient-centered outcomes (health-related quality of life). Pay for performance programs impacting surgical oncology patients focus primarily on preventing postoperative complications, but are not specific to cancer surgery. Future pay for performance programs in surgical oncology will likely focus on incentivizing high-quality, low-cost cancer care by evaluating process measures, patient-centered measures, and costs of care specific to cancer surgery.


The association between pulmonary vascular disease and respiratory improvement in infants with type I severe bronchopulmonary dysplasia.
To describe the association between echocardiographic measures of pulmonary vascular disease and time to respiratory improvement among infants with Type I severe bronchopulmonary dysplasia (sBPD). We measured the pulmonary artery acceleration time indexed to the right ventricular ejection time (PAAT/RVET) and right ventricular free wall longitudinal strain (RVFWLS) at 34-41 weeks' postmenstrual age. Cox-proportional hazards models were used to estimate the relationship between the PAAT/RVET, RVFWLS, and the outcome: days from 36 weeks' postmenstrual age to room-air or discharge with oxygen (≤0.5 L/min). For 102 infants, the mean PAAT/RVET and RVFWLS were 0.27 ± 0.06 and -22.63 ± 4.23%. An abnormal measurement was associated with an increased time to achieve the outcome (PAAT/RVET: 51v24, p < 0.0001; RVFWLS; 62v38, p = 0.0006). A normal PAAT/RVET was independently associated with a shorter time to outcome (aHR = 2.04, 1.11-3.76, p = 0.02). The PAAT/RVET may aid in anticipating timing of discharge in patients with type I severe BPD.


Capsular variation in experimental strains of Haemophilus influenzae.
This study sought to demonstrate that clinically untypable strains of Haemophilus influenzae are derivable from previously capsulated ones. Penicillin-induced forms were employed to explore in vivo and in vitro a possible mechanism of the reversible cycle vegetative to L-phase revertant. Normal H. influenzae type b (Rab), capsule-deficient strain ATCC 9333, and experimental L-phase and its revertants were used in this investigation. Capsular antigens, polyribose phosphate (PRP) content of each strain was assayed by orcinol and rocket immunoelectrophoretic methods. Intra- and inter-strain PRP differences were statistically analysed. Strain differences between in vivo and in vitro passaged extracts of strain 9333 and 9333 were significant (t-test P less than 0.01). There were also significant differences in vivo and in vitro between penicillin-treated, L-phase infected mouse isolates and penicillin-free, L-phase infected mouse isolates; and also between penicillin-treated, L-phase infected mouse isolates and revertant Rab infected mouse isolates (Mann-Whitney U-test P less than 0.02, P less than 0.01, respectively). These findings suggest that untypable isolates of H. influenzae are derivable from otherwise capsulated strains, depending on decapsulating factors in the microenvironment. Clinical implications of the findings are discussed.


Radiomics on Gadoxetate Disodium-enhanced MRI: Non-invasively Identifying Glypican 3-Positive Hepatocellular Carcinoma and Postoperative Recurrence.
To investigate the impact of preoperative gadoxetate disodium (EOB) MRI-based radiomics on predicting glypican 3 (GPC3)-positive expression and the relevant recurrence-free survival (RFS) of HCC ≤ 5 cm. Between January 2014 and October 2018, 259 patients with solitary HCC ≤ 5 cm who underwent hepatectomy and preoperative EOB-MRI were retrieved. Multivariate logistic regression was implemented to identify independent predictors for GPC3. By combining five feature selection strategies and three classifiers, 15 GPC3-oriented radiomics models could be constructed, the best of which with independent clinicoradiologic predictors was integrated into the comprehensive nomogram. GPC3 was an independent risk factor of postoperative recrudescence for HCC. Alpha-fetoprotein >20 ng/mL, homogenous T2 signal and hypointensity on hepatobiliary phase were independently related to GPC3-positive expression in the clinicoradiologic model. With 10 features selected by support vector machines-recursive feature elimination, logistic regression-based classifier achieved the best performance among 15 radiomics models. After five-fold cross-validation, our comprehensive nomogram acquired better average area under receiver operating characteristic curves (training and validation cohorts: 0.931 vs. 0.943) than the clinicoradiologic algorithm (0.738 vs. 0.739) and the optimal radiomics model (0.943 vs. 0.931). Net reclassification indexes further demonstrated the superiority of GPC3 nomogram over clinicoradiologic and radiomics algorithms (46.54%, p < 0.001; 7.84%, p = 0.207). Meanwhile, higher radiomics score significantly shortened the median RFS (from >77.9 to 48.2 months, p = 0.044), which was analogue to that of the histological GPC3-positive phenotype (from >73.9 to 43.2 months, p < 0.001). Preoperative EOB-MRI radiomics-based nomogram satisfactorily distinguished GPC3 status and outcomes of solitary HCC ≤ 5 cm.


Intracellular metabolism and enzymatic phosphorylation of 9-(1,3-dihydroxy-2-propoxymethyl)guanine and acyclovir in herpes simplex virus-infected and uninfected cells.
The antiherpes agent 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG) is a much more potent inhibitor of herpes simplex viruses in vivo than acyclovir, yet both are equally active in vitro against these viruses. To explain this difference, studies were conducted to compare the intracellular metabolism and enzymatic phosphorylation of the two compounds. In herpes type 1 and type 2 infected cells, the levels of DHPG triphosphate were only about 2-fold greater than levels of acyclovir triphosphate at virus-inhibitory concentrations (less than or equal to microM). At concentrations greater than 2.5 microM in herpes type 1 but not in type 2 infected cells, acyclovir phosphorylation was inhibited relative to that of DHPG. When drug was removed after 6 hr from infected cells, acyclovir triphosphate rapidly degraded to acyclovir and was excreted into the culture medium. In contrast, DHPG triphosphate persisted at 60-70% of the original level for 18 hr after drug removal, and DHPG excretion from cells was very slow. This finding could be a key factor to the superior potency of DHPG in animals, despite the fact that blood levels of both compounds fall rapidly after dosing. In uninfected cells, low levels of DHPG and acyclovir triphosphates were produced at 100 microM concentrations. Phosphorylation of DHPG to mono-, di- and triphosphates by purified viral and cell enzymes was more rapid than that of acyclovir. However, acyclovir triphosphate was a much more potent inhibitor of herpes virus and cell DNA polymerases.


Calretinin and parvalbumin immunoreactive interneurons in the retrosplenial cortex of the rat brain: Qualitative and quantitative analyses.
The retrosplenial cortex (RSC) is a mesocortical region broadly involved with memory and navigation. It shares many characteristics with the perirhinal cortex (PRC), both of which appear to be significantly involved in the spreading of epileptic activity. We hypothesized that RSC possesses an interneuronal composition similar to that of PRC. To prove the hypothesis we studied the general pattern of calretinin (CR) and parvalbumin (PV) immunoreactivity in the RSC of the rat brain, its optical density as well as the morphological features and density of CR- and PV-immunoreactive (CR+ and PV+) interneurons. We also analyzed the overall neuronal density on Nissl-stained sections in RSC. Finally, we compared our results with our earlier analysis of PRC (Barinka et al., 2012). Compared to PRC, RSC was observed to have a higher intensity of PV staining and lower intensity of CR staining of neuropil. Vertically-oriented bipolar neurons were the most common morphological type among CR+ neurons. The staining pattern did not allow for a similarly detailed analysis of somatodendritic morphology of PV+ neurons. RSC possessed lower absolute (i.e., neurons/mm(3)) and relative (i.e., percentage of the overall neuronal population) densities of CR+ neurons and similar absolute and lower relative densities of PV+ neurons relative to PRC. CR: PV neuronal ratio in RSC (1:2 in area 29 and 1:2.2 in area 30) differed from PRC (1:1.2 in area 35 and 1:1.7 in area 36). In conclusion, RSC, although similar in many aspects to PRC, differs strikingly in the interneuronal composition relative to PRC.


Mineralogy and phase transition mechanisms of atmospheric mineral particles: Migration paths, sources, and volatile organic compounds.
Inorganic mineral particles play an important role in the formation of atmospheric aerosols in the Sichuan Basin. Atmospheric haze formation is accompanied by the phase transition of mineral particles under high humidity and stable climatic conditions. Backward trajectory analysis was used in this study to determine the migration trajectory of atmospheric mineral particles. Furthermore, Positive matrix factorization (PMF) was used to analyze the sources of atmospheric mineral particles. The phase transition mechanisms of atmospheric mineral particles were studied using ion chromatography, inductively coupled plasma emission spectrometry, total organic carbon analysis, X-ray diffraction, Fourier-transform infrared spectroscopy, scanning electron microscopy coupled with energy dispersive spectrometry, and grand canonical Monte Carlo methods. Three migration and phase transition paths were identified for the mineral particles. Sources of atmospheric mineral particles included combustion, vehicle emissions, industrial emissions, agricultural sources, and mineral dust. The main mineral phases in atmospheric particles, calcite and dolomite, were transformed into gypsum, and muscovite may be transformed into kaolinite. The phase transition of mineral particles seriously affects the formation of aerosols and worsens haze. Typically, along the Nanchong-Suining-Neijiang-Zigong-Yibin path, calcite is converted into gypsum under the influence of man-made inorganic pollution gases, which worsen the haze conditions and cause slight air pollution for 3-5 days. However, along the Guangyuan-Mianyang-Deyang-Chengdu-Meishan-Ya'an path, anthropogenic volatile organic compounds (VOCs) hindered gypsum formation from dolomite. Furthermore, dolomite and VOCs formed stable adsorption systems (system energies from -0.41 to -4.76 eV, long bonds from 0.20 to 0.24 nm). The adsorption system of dolomite and m/p-xylene, with low system energy (-1.46 eV/-1.33 eV) and significant correlation (r2 = 0.991, p < 0.01), was the main cause of haze formation. Consequently, calcite gypsification and dolomite--VOC synergism exacerbated regional haze conditions. This study provides a theoretical reference for the mechanism of aerosol formation in basin climates.


[The effect of the I/D polymorphism of the angiotensin-converting enzyme gene on the cardiovascular risk and graft survival of kidney transplant patients].
Renal transplantation provides longer life expectancy in patients with renal failure. Nonetheless, this improved life expectancy is still shorter than that for the general population. The main couse of death in renal transplant patients is cardiovascular disease, and chronic allograft nephropathy is the most significant cause of graft loss. Genetic polymorphisms of the renin angiotensin system have been implicated in both chronic allograft nephropathy and fatal cardiovascular diseases. The long term goal of the authors was to improve the survival of renal transplanted patients. The authors aimed to identify novel biomarkers which correlate with the survival of the transplant organ and the recipient with a special attention to elements of the renin-angiotensin system. A retrospective clinical trial was performed involving 72 renal transplanted patients. Angiotensin-converting enzyme I/D genotypes and activity, kidney function and morphological properties of the heart were determined. A significant positive correlation was found between the DD genotype of the angiotensin-converting enzíme gene, and the DD genotype predicted severe left ventricular hypertrophy. These findings suggest that the I/D genotypes of the angiotensin-converting enzyme gene predict not only the expected survival of the transplanted organ, but also that of the patient. Patients with the DD genotype are more susceptible for transplant failure. These patients should be identified and a special attention should be made on their pharmacological treatment (renin-angiotensin system inhibition), and their complience should also be maintained.


Eosinophil-dependent cytotoxicity in rat schistosomiasis. Involvement of IgG2a antibody and role of mast cells.
Using Schistoma mansoni schistosomula collected in vitro and sensitized with 42-day-infected rat serum, normal rat peritoneal cells enriched in eosinophils were proven to subsequently adhere to and kill the schistosomula within 24 to 48 h. The cell-dependent, heat-stable antibody in infected rat serum reached a peak between 30 and 42 days after infection. Inhibition experiments with aggregated immunoglobulins indicated the role of IgG2a antibody in the adherence of eosinophils to sensitized schistosomula. The immune absorption technique showed that IgG2a antibody was involved in the mechanism of cytotoxicity of effector to target cells, whereas the role of IgE antibody could be excluded. Ultrastructural studies revealed the constant presence of eosinophils and mast cells in contact with schistosomula. The use of purified cell populations showed that the cytotoxic effect of the original cell population was significantly decreased after depletion of mast cells and partially restored after addition of mast cells. These observations, together with those concerning the role of IgE immune complexes in macrophage cytotoxicity, suggest the possible participation of anaphylactic antibodies in immunity to schistosomes in the rat.


Autosampler for portable capillary electrophoresis.
A compact, inexpensive sampler instrument for portable capillary electrophoresis was developed for monitoring illegal drugs in human body fluids and evaluated for γ-hydroxybutyric acid (GHB) in simulated saliva samples. The sampler uses peristaltic pumps and pinch and check valves to activate liquid flows. This short communication addresses aspects of CE associated with portability, open access instrumentation, and prospects of CE for citizen science. The extensive use of items provided by the electronic and computer industry contributes to this trend.


Membrane-disruptive abilities of beta-hairpin antimicrobial peptides correlate with conformation and activity: a 31P and 1H NMR study.
The membrane interaction and solution conformation of two mutants of the beta-hairpin antimicrobial peptide, protegrin-1 (PG-1), are investigated to understand the structural determinants of antimicrobial potency. One mutant, [A(6,8,13,15)] PG-1, does not have the two disulfide bonds in wild-type PG-1, while the other, [Delta(4,18) G10] PG-1, has only half the number of cationic residues. 31P solid-state NMR lineshapes of uniaxially aligned membranes indicate that the membrane disorder induced by the three peptides decreases in the order of PG-1 > [Delta(4,18) G10] PG-1>>[A(6,8,13,15)] PG-1. Solution NMR studies of the two mutant peptides indicate that [Delta(4,18) G10] PG-1 preserves the beta-hairpin fold of the wild-type peptide while [A(6,8,13,15)] PG-1 adopts a random coil conformation. These NMR results correlate well with the known activities of these peptides. Thus, for this class of peptides, the presence of a beta-hairpin fold is more essential than the number of cationic charges for antimicrobial activity. This study indicates that 31P NMR lineshapes of uniaxially aligned membranes are well correlated with antimicrobial activity, and can be used as a diagnostic tool to understand the peptide-lipid interactions of these antimicrobial peptides.


Development of Clinical Pharmacy services at King Khalid University Hospital and its impact on the quality of healthcare provided.
Clinical Pharmacy is a unique service provided by the leading pharmacy departments in the United States. The concept of Clinical Pharmacy evolved after the significant increase in number of pharmaceuticals in the market and the increasing potential of drug interactions. However, the Clinical Pharmacist is not merely an individual who advises on drug interactions. There are a number of functions which include but are not limited to; the design of appropriate drug therapy, such as Pharmacokinetic assessment and evaluation to optimize drug therapy, drug information dissemination to the physicians and other healthcare providers and participation as a toxicology consultant in Poison management. At the King Khalid University Hospital (KKUH) the first Clinical Pharmacy services program began in 1983. The aim of this study is to evaluate the impact of our Clinical Pharmacy program on the patients' care as well as its perception by the Medical staff that came from different parts of the world. Our Clinical Pharmacists were asked to record any suggestions or interventions in the form. The forms were all collected at the end of each day and entered into a database for analysis. Each intervention was analyzed in order to assess the merit of the action in terms of the therapeutic, financial and direct cost impact. The study showed a positive impact on the patients' care as well as on the economy of the drugs prescribed. Meanwhile, the service was very much appreciated by the Medical staff as well as other healthcare providers.


Glucocorticoid inhibition of Na-Pi cotransport in renal epithelial cells is mediated by protein kinase C.
The effect and mechanism of action of glucocorticoids (GC) on Na-Pi cotransport were evaluated in opossum kidney cells. Dexamethasone (1-1000 nM) inhibited sodium-dependent Pi uptake in a time- and concentration-dependent manner. Inhibition was maximal after a 6-h incubation with dexamethasone and was prevented by cycloheximide and actinomycin D. The effect was related to a 37% decrease of the Vmax value after incubation with 100 nM dexamethasone. The effect of dexamethasone was mimicked by cortisol and blocked by GC receptor antagonists RU38486 and progesterone. GC affected neither glucose or alanine uptake nor Na/H exchange activity. Inhibition of Pi uptake persisted when Na/H was blocked by amiloride or dimethylamiloride. GC had no effect on basal or parathyroid hormone- and forskolin-stimulated intracellular cAMP content. Dexamethasone and extracellular cAMP, parathyroid hormone, or 3-isobutyl-1-methylxanthine had additive inhibitory effects on Pi uptake. Staurosporine, GF109203X, or calphostin C (three dissimilar inhibitors of protein kinase C (PKC)) and PKC down-regulation blunted the inhibitory effect of glucocorticoids on Pi uptake. GC increased both membrane-bound PKC activity and the membrane/cytosol PKC activity ratio. This is the first report of GC activation of PKC in renal cells, which appears to mediate the steroid inhibitory effect on Pi transport.


Differences in transmission through the dorsal column nuclei in spontaneously hypertensive and Wistar Kyoto rats.
The medical lemniscal evoked potential in response to a range of footshock intensities was recorded in spontaneously hypertensive (SHR) and Wistar Kyoto rats (WKY). Input-output (I-O) relationships were constructed as the percent of maximum response at each intensity. The SHR had a steeper I-O relationship than did the WKY. This difference was also evident when SHR maintained normotensive from weaning with hydralazine were compared with identically treated WKY. The treatment itself steepened the I-O relationship of the SHR while leaving that of the WKY unchanged. These results indicate an inherent hyperresponsiveness in the SHR dorsal column nuclei and an inhibitory effect of elevated blood pressure on transmission through these nuclei.


Nonlinear self-dual network equations: modulation instability, interactions of higher-order discrete vector rational solitons and dynamical behaviours.
The nonlinear self-dual network equations that describe the propagations of electrical signals in nonlinear LC self-dual circuits are explored. We firstly analyse the modulation instability of the constant amplitude waves. Secondly, a novel generalized perturbation (M, N - M)-fold Darboux transform (DT) is proposed for the lattice system by means of the Taylor expansion and a parameter limit procedure. Thirdly, the obtained perturbation (1, N - 1)-fold DT is used to find its new higher-order rational solitons (RSs) in terms of determinants. These higher-order RSs differ from those known results in terms of hyperbolic functions. The abundant wave structures of the first-, second-, third- and fourth-order RSs are exhibited in detail. Their dynamical behaviours and stabilities are numerically simulated. These results may be useful for understanding the wave propagations of electrical signals.


When DNA repair goes wrong: BER-generated DNA-protein crosslinks to oxidative lesions.
Free radicals generate an array of DNA lesions affecting all parts of the molecule. The damage to deoxyribose receives less attention than base damage, even though the former accounts for ∼20% of the total. Oxidative deoxyribose fragments (e.g., 3'-phosphoglycolate esters) are removed by the Ape1 AP endonuclease and other enzymes in mammalian cells to enable DNA repair synthesis. Oxidized abasic sites are initially incised by Ape1, thus recruiting these lesions into base excision repair (BER) pathways. Lesions such as 2-deoxypentos-4-ulose can be removed by conventional (single-nucleotide) BER, which proceeds through a covalent Schiff base intermediate with DNA polymerase β (Polβ) that is resolved by hydrolysis. In contrast, the lesion 2-deoxyribonolactone (dL) must be processed by multinucleotide ("long-patch") BER: attempted repair via the single-nucleotide pathway leads to a dead-end, covalent complex with Polβ cross- linked to the DNA by an amide bond. We recently detected these stable DNA-protein crosslinks (DPC) between Polβ and dL in intact cells. The features of the DPC formation in vivo are exactly in keeping with the mechanistic properties seen in vitro: Polβ-DPC are formed by oxidative agents in line with their ability to form the dL lesion; they are not formed by non-oxidative agents; DPC formation absolutely requires the active-site lysine-72 that attacks the 5'-deoxyribose; and DPC formation depends on Ape1 to incise the dL lesion first. The Polβ-DPC are rapidly processed in vivo, the signal disappearing with a half-life of 15-30min in both mouse and human cells. This removal is blocked by inhibiting the proteasome, which leads to the accumulation of ubiquitin associated with the Polβ-DPC. While other proteins (e.g., topoisomerases) also form DPC under these conditions, 60-70% of the trapped ubiquitin depends on Polβ. The mechanism of ubiquitin targeting to Polβ-DPC, the subsequent processing of the expected 5'-peptidyl-dL, and the biological consequences of unrepaired DPC are important to assess. Many other lyase enzymes that attack dL can also be trapped in DPC, so the processing mechanisms may apply quite broadly.


Evaluating the prebiotic effect of oligosaccharides on gut microbiome wellness using in vitro fecal fermentation.
We previously proposed the Gut Microbiome Wellness Index (GMWI), a predictor of disease presence based on a gut microbiome taxonomic profile. As an application of this index for food science research, we applied GMWI as a quantitative tool for measuring the prebiotic effect of oligosaccharides. Mainly, in an in vitro anaerobic batch fermentation system, fructooligosaccharides (FOS), galactooligosaccharides (GOS), xylooligosaccharides (XOS), inulin (IN), and 2'-fucosyllactose (2FL), were mixed separately with fecal samples obtained from healthy adult volunteers. To find out how 24 h prebiotic fermentation influenced the GMWI values in their respective microbial communities, changes in species-level relative abundances were analyzed in the five prebiotics groups, as well as in two control groups (no substrate addition at 0 h and for 24 h). The GMWI of fecal microbiomes treated with any of the five prebiotics (IN (0.48 ± 0.06) > FOS (0.47 ± 0.03) > XOS (0.33 ± 0.02) > GOS (0.26 ± 0.02) > 2FL (0.16 ± 0.06)) were positive, which indicates an increase of relative abundances of microbial species previously found to be associated with a healthy, disease-free state. In contrast, the GMWI of samples without substrate addition for 24 h (-0.60 ± 0.05) reflected a non-healthy, disease-harboring microbiome state. Compared to the original prebiotic index (PI) and α-diversity metrics, GMWI provides a more data-driven, evidence-based indexing system for evaluating the prebiotic effect of food components. This study demonstrates how GMWI can be applied as a novel PI in dietary intervention studies, with wider implications for designing personalized diets based on their impact on gut microbiome wellness.


Psychometric properties and measurement invariance of the 7-item General Anxiety Disorder scale (GAD-7) in an Italian coronary heart disease population.
Generalized Anxiety Disorder is predominant in coronary heart disease (CHD) patients. 7-item Generalized Anxiety Disorder (GAD-7) scale psychometric properties have never been tested in CHD populations. This study aims to verify the GAD-7 psychometric properties and measurement invariance in an Italian CHD sample. A baseline data secondary analysis from HEARTS-IN-DYADS study. Several healthcare facilities enrolled an adult inpatient sample. Anxiety and depression data were collected using GAD-7 and Patient Health Questionnaire 9 (PHQ-9). Factorial validity was assessed with confirmatory factor analysis; construct validity was tested by correlating GAD-7 scores with PHQ-9 scores and other sociodemographic characteristics; internal consistency reliability was assessed using Cronbach's alpha and the composite reliability index, while confirmatory multigroup factor analysis was employed to investigate measurement invariance across gender and age (65 vs. 65). We enrolled 398 patients (mean age 64.7 years; 78.9 % male; 66.8 % married). Factor structure was confirmed as unidimensional. Construct validity was confirmed with significant associations between GAD-7 and PHQ-9 scores, female gender, having a caregiver, and being employed. Cronbach's alpha and composite reliability index were 0.89 and 0.90, respectively. Measurement invariance across gender and age was confirmed at the scalar level. A convenience sample in one European country, a small female sample, validity testing against a single criterion. Study results demonstrate adequate validity and reliability of the GAD-7 in an Italian CHD sample. It showed satisfactory invariance properties; GAD-7 is suitable for measuring anxiety in CHD while making significant comparisons of scores among stratified gender and age groups.


Increased expression of 11beta-hydroxysteroid dehydrogenase type 1 in type 2 diabetic myotubes.
Alterations in glucocorticoid hormone metabolism in skeletal muscle have been suggested to contribute to the pathogenesis of the metabolic syndrome. Circulating glucocorticoids consist of inactive cortisone and active cortisol interconverted in various tissues by the enzyme 11beta hydroxysteroid dehydrogenase (HSD). This study aims to investigate whether human myotubes established from healthy obese and matched obese type 2 diabetic (T2D) subjects reveal differences in the expression level of glucocorticoid receptor (GR) and 11beta hydroxysteroid dehydrogenase (HSD1 and HSD2), and to investigate whether chronic exposure to cortisone affects glucose transport. In myotubes established from T2D and healthy control subjects we determined the mRNA expression of HSD1, HSD2, GR and determined basal and insulin-stimulated glucose uptake in myotubes precultured with cortisone, cortisol and the HSD1 inhibitor, carbenoxolone for four days. Myotubes established from T2D subjects showed an increased expression of HSD1 mRNA, but with no differences in mRNA of GRalpha, LXRalpha and LXRbeta, whereas HSD2 mRNA was not expressed. Cortisone reduced glucose uptake in diabetic myotubes and the cortisone effect could be abolished by the HSD1 inhibitor carbenoxolone. Our study shows that cortisone reduces glucose uptake in diabetic myotubes and that this effect seems mediated by an increased mRNA HSD1 expression emphasizing that the local conversion of inactive to active glucocorticoids may be important in the pathogenesis of insulin resistance.


A review on the production of nitrogen-containing compounds from microalgal biomass via pyrolysis.
Nitrogen-containing compounds (NCCs) which may be produced from nitrogen-rich biomass such as microalgae, may find important biochemical and biomedical applications. This review summarizes the recent knowledge about the formation mechanism of NCCs during pyrolysis of microalgae. The key technical and biological aspects of microalgae and pyrolysis process parameters, which influence the formation of NCCs, have been analyzed. The mechanism of formation of NCCs such as indole, pyridine, amides, and nitriles during primary and secondary pyrolysis reactions are elaborated. It has been emphasized that the pyrolysis conditions and the use of catalysts had significant impacts on the yields and compositions of NCCs. The available information shows that the transformation of nitrogen and nitrogen functionalities during pyrolysis are strongly associated with the formation process of NCCs. The challenges in the development of pyrolysis technologies for the production of NCCs from microalgae are identified with future research needs identified.


Meta-analysis derived atopic dermatitis (MADAD) transcriptome defines a robust AD signature highlighting the involvement of atherosclerosis and lipid metabolism pathways.
Atopic dermatitis (AD) is a common inflammatory skin disease with limited treatment options. Several microarray experiments have been conducted on lesional/LS and non-lesional/NL AD skin to develop a genomic disease phenotype. Although these experiments have shed light on disease pathology, inter-study comparisons reveal large differences in resulting sets of differentially expressed genes (DEGs), limiting the utility of direct comparisons across studies. We carried out a meta-analysis combining 4 published AD datasets to define a robust disease profile, termed meta-analysis derived AD (MADAD) transcriptome. This transcriptome enriches key AD pathways more than the individual studies, and associates AD with novel pathways, such as atherosclerosis signaling (IL-37, selectin E/SELE). We identified wide lipid abnormalities and, for the first time in vivo, correlated Th2 immune activation with downregulation of key epidermal lipids (FA2H, FAR2, ELOVL3), emphasizing the role of cytokines on the barrier disruption in AD. Key AD "classifier genes" discriminate lesional from nonlesional skin, and may evaluate therapeutic responses. Our meta-analysis provides novel and powerful insights into AD disease pathology, and reinforces the concept of AD as a systemic disease.


Epidemic versus economic performances of the COVID-19 lockdown: A big data driven analysis.
Lockdown measures have been a "panacea" for pandemic control but also a violent "poison" for economies. Lockdown policies strongly restrict human mobility but mobility reduce does harm to economics. Governments meet a thorny problem in balancing the pros and cons of lockdown policies, but lack comprehensive and quantified guides. Based on millions of financial transaction records, and billions of mobility data, we tracked spatio-temporal business networks and human daily mobility, then proposed a high-resolution two-sided framework to assess the epidemiological performance and economic damage of different lockdown policies. We found that the pandemic duration under the strictest lockdown is less about two months than that under the lightest lockdown, which makes the strictest lockdown characterize both epidemiologically and economically efficient. Moreover, based on the two-sided model, we explored the spatial lockdown strategy. We argue that cutting off intercity commuting is significant in both epidemiological and economical aspects, and finally helped governments figure out the Pareto optimal solution set of lockdown strategy.


The Individual and Societal Burden of Treatment-Resistant Depression: An Overview.
Major depressive disorder is characterized by depressed mood and/or anhedonia with neurovegetative symptoms and neurocognitive changes affecting an individual's functioning in multiple aspects of life. Treatment outcomes with commonly used antidepressants remain suboptimal. Treatment-resistant depression (TRD) should be considered after inadequate improvement with two or more antidepressant treatments of adequate dose and duration. TRD has been associated with increased disease burden including higher associated costs (both socially and financially) affecting both the individual and society. Additional research is needed to better understand the long-term burden of TRD to both the individual and society.


Recurrence with histological transformation 40 days after autologous peripheral blood stem cell transplantation (APBSCT) for cutaneous CD30-negative large T cell lymphoma.
Localized cutaneous nontender nodules appeared on the back of a 52-year-old Japanese woman. Skin biopsy revealed atypical large T-lymphocytes infiltrating the dermis. CD30 staining was negative in tumor cells. The diagnosis was CD30-negative cutaneous large T cell lymphoma. There was no evidence of peripheral lymphadenopathy or bone marrow involvement. Six cycles of induction chemotherapy were administered and a complete clinical remission (CCR) was attained. Local irradiation was not given. As the clinical course of CD30-negative cutaneous large T cell lymphoma is recurrent and often incurable with conventional chemoradiotherapy, she received high-dose chemotherapy without total body irradiation (TBI) followed by unpurged autologous peripheral blood stem cell transplantation (APBSCT). A relapse in the skin followed 40 days after APBSCT, but tumor cells transformed into a CD30-positive anaplastic large cell lymphoma (ALCL). We question the need for TBI in conditioning and for purged stem cells for APBSCT in patients with high risk cutaneous lymphomas.


Prostaglandin E2 Leads to the Acquisition of DNMT3A-Dependent Tolerogenic Functions in Human Myeloid-Derived Suppressor Cells.
Myeloid-derived suppressor cells (MDSCs) and dendritic cells (DCs) arise from common progenitors. Tumor-derived factors redirect differentiation from immune-promoting DCs to tolerogenic MDSCs, an immunological hallmark of cancer. Indeed, in vitro differentiation of DCs from human primary monocytes results in the generation of MDSCs under tumor-associated conditions (PGE2 or tumor cell-conditioned media). Comparison of MDSC and DC DNA methylomes now reveals extensive demethylation with specific gains of DNA methylation and repression of immunogenic-associated genes occurring in MDSCs specifically, concomitant with increased DNA methyltransferase 3A (DNMT3A) levels. DNMT3A downregulation erases MDSC-specific hypermethylation, and it abolishes their immunosuppressive capacity. Primary MDSCs isolated from ovarian cancer patients display a similar hypermethylation signature in connection with PGE2-dependent DNMT3A overexpression. Our study links PGE2- and DNMT3A-dependent hypermethylation with immunosuppressive MDSC functions, providing a promising target for therapeutic intervention.


Usefulness of diffusion-weighted imaging and the apparent diffusion coefficient in the assessment of head and neck tumors.
The aim of this review was to determine the usefulness of diffusion-weighted imaging (DWI) and the apparent diffusion coefficient (ADC) in the assessment of head and neck tumors. DWI and the ADC can help in the differential diagnosis of particular disorders (such as carcinomas vs lymphomas, or necrosis vs abscess) of the head and neck. The ADC can also provide further information to help differentiate benign from malignant tumors, as ADC values are usually lower in cases of malignancy. However, the ADC is itself influenced by various complex factors such as the cellularity and matrix of tumors and there is also some overlap between certain benign and malignant tumors of the salivary glands.


Effect of live Eimeria vaccination or salinomycin on growth and immune status in broiler chickens receiving in-feed inclusion of gelatin and vitamin E.
This experiment determined if 2% of gelatin, to improve the levels of proline and glycine in the diet, and 70 mg/kg of vitamin E supplementation would relieve the impaired performance of male Cobb broilers vaccinated for coccidiosis. Half of the chicks were vaccinated via water (live oocysts), while the other half received medication (salinomycin) in the feed until 35 d of age. The effects of coccidiosis vaccine on performance and mRNA levels of genes involved in mucin synthesis, cytokines, trefoil family factor-2 (TFF2), and metabolic processes (CD36) in the jejunum of broilers were measured. Vaccination negatively affected performance in the first 21 d; however, the inclusion of gelatin and vitamin E reduced this negative response. Additionally, supplementation with these nutrients led to an improvement in broilers receiving the coccidiostat (P < 0.05). From 21 to 35 d, birds treated with gelatin and coccidiosis vaccine experienced better body weight gain than birds without gelatin and vitamin E (P < 0.05). Vaccinated chickens had decreased body weight and decreased anti-inflammatory cytokine expression. Furthermore, they had increased inflammatory cytokine expression, mucin 2 expression, and TFF2 compared to salinomycin-fed broilers (P < 0.05). Transcripts for IL-1B, IFN-y, MUC2, TFF2 were decreased while mRNAs for IL-4 and IL-10 increased in salinomycin-fed broilers compared to vaccinated broilers (P < 0.05). In conclusion, broilers vaccinated against coccidiosis increase their pro-inflammatory immune status and mucin expression compared to broilers receiving salinomycin. These events may contribute to lower performance in vaccinated broiler chicks. Moreover, vitamin E and gelatin can minimize the vaccine's negative immune effects and promote better performance.


RNA editing of a conserved reading frame in plant mitochondria increases its similarity to two overlapping reading frames in Escherichia coli.
An open reading frame (orfx) in mitochondria of the higher plants Oenothera berteriana and Arabidopsis thaliana is homologous to orf244 in the mitochondrial genome of Marchantia polymorpha. Homologous sequences are also present in carrot, potato and sugar beet. Profile analysis revealed similarity to two overlapping reading frames in the Escherichia coli genome. Potential translation initiation at conserved ATA (isoleucine) and TTG (leucine) codons is discussed. Transcripts of the open reading frame are altered by RNA editing in Arabidopsis and Oenothera downstream of these codons, suggesting this to be the functionally important region.


Slipped capital femoral epiphysis associated with Rubinstein-Taybi syndrome.
The association of Rubinstein-Taybi syndrome (RTS) and slipped capital femoral epiphysis (SCFE) is described in a girl aged 9 years and 10 months. SCFE has never been reported associated with RTS, neither as an isolated anomaly, nor in a familial pedigree. However, a "stiff gait" is frequently described in RTS patients and, furthermore, obesity is a frequent feature of RTS patients. Some reports in the literature suggest the need for an early diagnosis of SCFE among adolescent relatives of patients with SCFE. Since many SCFEs are asymptomatic and an early diagnosis is essential for a favorable prognosis, we suggest an annual echotomographic or radiological examination of the hips in RTS patients.


Comparative proteomics of thylakoids from Arabidopsis grown in laboratory and field conditions.
Compared to controlled laboratory conditions, plant growth in the field is rarely optimal since it is frequently challenged by large fluctuations in light and temperature which lower the efficiency of photosynthesis and lead to photo-oxidative stress. Plants grown under natural conditions therefore place an increased onus on the regulatory mechanisms that protect and repair the delicate photosynthetic machinery. Yet, the exact changes in thylakoid proteome composition which allow plants to acclimate to the natural environment remain largely unexplored. Here, we use quantitative label-free proteomics to demonstrate that field-grown Arabidopsis plants incorporate aspects of both the low and high light acclimation strategies previously observed in laboratory-grown plants. Field plants showed increases in the relative abundance of ATP synthase, cytochrome b 6 f, ferredoxin-NADP+ reductases (FNR1 and FNR2) and their membrane tethers TIC62 and TROL, thylakoid architecture proteins CURT1A, CURT1B, RIQ1, and RIQ2, the minor monomeric antenna complex CP29.3, rapidly-relaxing non-photochemical quenching (qE)-related proteins PSBS and VDE, the photosystem II (PSII) repair machinery and the cyclic electron transfer complexes NDH, PGRL1B, and PGR5, in addition to decreases in the amounts of LHCII trimers composed of LHCB1.1, LHCB1.2, LHCB1.4, and LHCB2 proteins and CP29.2, all features typical of a laboratory high light acclimation response. Conversely, field plants also showed increases in the abundance of light harvesting proteins LHCB1.3 and CP29.1, zeaxanthin epoxidase (ZEP) and the slowly-relaxing non-photochemical quenching (qI)-related protein LCNP, changes previously associated with a laboratory low light acclimation response. Field plants also showed distinct changes to the proteome including the appearance of stress-related proteins ELIP1 and ELIP2 and changes to proteins that are largely invariant under laboratory conditions such as state transition related proteins STN7 and TAP38. We discuss the significance of these alterations in the thylakoid proteome considering the unique set of challenges faced by plants growing under natural conditions.


A Three-Dimensional Study of the Nasolabial Soft Tissue Symmetry in Children With Unilateral Complete Cleft Lip and Palate Using Traditional and Split-Type Nasoalveolar Molding.
Presurgical nasoalveolar molding (NAM) is the most common preoperative treatment for cleft lip and palate. However, NAM may have some limitations such as requiring high technical sensitivity and frequent visits. To simplify the device, some scholars have changed the traditional NAM into a split-NAM consisting of a alveolar molding plate and a nasal hook. This study compared the shaping effect of split NAM and traditional NAM on nasolabial soft tissue using three-dimensional (3D) measurement. A total of 39 patients with unilateral cleft lip and palate (UCLP) were enrolled and divided into 2 groups. 13 patients were treated with split-NAM while the other 26 patients were treated with traditional NAM. 3D images of all patients' craniofacial soft tissue before and after NAM treatment were recorded and measured by three-dimensional software. Statistical analysis of measurements in both groups was performed using SPSS software. After treatment, nasal soft tissue symmetry in the split-NAM group was better improved than that in the NAM group in vertical and anterior-posterior direction, but was worse improved in transverse direction. There was no significant difference in labial soft tissue symmetry between two groups. The split NAM can better elevate the alar and nostrils of the cleft side, and have a better forward effect on alar outer edge, nasal base, and nostrils. However, the traditional NAM can better reduce the width of nasal base.


Physical inactivity and associated factors among university students in 23 low-, middle- and high-income countries.
The aim of this study was to determine estimates of the prevalence and social correlates of physical inactivity among university students in 23 low-, middle- and high-income countries. The International Physical Activity Questionnaire was used to collect data from 17,928 undergraduate university students (mean age 20.8, SD = 2.8) from 24 universities in 23 countries. The prevalence of physical inactivity was 41.4 %, ranging from 21.9 % in Kyrgyzstan to 80.6 % in Pakistan. In multivariate logistic regression, older age (22-30 years), studying in a low- or lower middle-income country, skipping breakfast and lack of social support were associated with physical inactivity. In men, being underweight, being overweight or obese, not avoiding fat and cholesterol, not having severe depression symptoms, low beliefs in the health benefits of physical activity, low personal control and knowledge of exercise-heart link, and in women, not trying to eat fibre, low personal mastery and medium personal control were additionally associated with physical inactivity. Four in each ten students are physically inactive, calling for strategic interventions by relevant professionals in higher educational institutions.


Efficacy study of two novel hyaluronic acid-based formulations for viscosupplementation therapy in an early osteoarthrosic rabbit model.
Viscosupplementation (VS) is a therapy for osteoarthrosis (OA) consisting of repetitive intra-articular injections of hyaluronic acid (HA). It is known to be clinically effective in relieving pain and increasing joint mobility by restoring joint homeostasis. In this study, the effects of two novel HA-based VS hydrogel formulations were assessed and challenged against a pure HA commercial formulation for the first time and this in a rabbit model of early OA induced by anterior cruciate ligament transection (ACLT). The first formulation tested was a hybrid hydrogel composed of HA and reacetylated chitosan, a biopolymer considered to be chondroprotective, assembled thanks to an ionic shielding. The second formulation consisted of a novel HA polymer grafted with antioxidant molecules (HA-4AR) aiming at decreasing OA oxidative stress and increasing HA retention time in the articulation. ACLT was performed on rabbits in order to cause structural changes comparable to traumatic osteoarthrosis. The protective effects of the different formulations were observed on the early phase of the pathology in a full randomized and blinded manner. The cartilage, synovial membrane, and subchondral bone were evaluated by complementary investigation techniques such as gross morphological scoring, scanning electron microscopy, histological scoring, and micro-computed tomography were used. In this study, ACLT was proven to successfully reproduce early OA articular characteristics found in humans. HA and HA-4AR hydrogels were found to be moderately protective for cartilage as highlighted by μCT. The HA-4AR was the only formulation able to decrease synovial membrane hypertrophy occurring in OA. Finally, the hybrid HA-reacetylated chitosan hydrogel surprisingly led to increased subchondral bone remodeling and cartilage defect formation. This study shows significant effects of two innovative HA modification strategies in an OA rabbit model, which warrant further studies toward more effective viscosupplementation formulations.


Inflammatory Markers in the Diagnosis of Fibromyalgia.
Fibromyalgia syndrome (FMS) is a chronic disorder characterized by widespread musculoskeletal pain accompanied by various additional symptoms. The prevalence of FMS ranges between 2-8% of the population. The exact pathophysiology of the disease remains unknown, and under certain circumstances it is difficult for the physician to diagnose. Previous studies have shown a correlation between inflammatory biomarkers such as C-reactive protein (CRP) and FMS activity, suggesting that an inflammatory component may play a role in this disease pathogenesis. To investigate the role of certain new inflammatory biomarkers in the diagnosis of patients with FMS. In this study data were collected from FMS patients who were admitted to Ziv Medical Center during the period 2013 to 2019 in an attempt to find a connection between inflammatory markers detectable by a traditional complete blood count (CBC) tests such as neutrophil-lymphocytes ratio (NLR), platelet-lymphocyte ratio (PLR), mean platelet value (MPV), red cell distribution width (RDW), and C-reactive protein (CRP) and FMS. We found significantly higher CRP levels, MPV, and PLR and lower lymphocyte count in the FMS group compared to the control group. FMS has certain inflammatory components that may be useful in disease diagnosis.


Location-independent determination of the activity of a point source in absorbing media.
The unknown distribution of a radioactive material in a bulky volume will induce considerable inaccuracies in the determination of its activity due to the unknown geometry and absorption factors. The methods of using two detectors and of employing the full-energy peak to total ratio were investigated to evaluate parameters, which can be used to improve the accuracy of activity determination.


Clinico-epidemiological profile and predictors of severe illness in young infants (0-59 days) in Ghana.
Young infant mortality has remained high and relatively unchanged compared with deaths of older infants. Strategies to reduce infant mortality, however, are mostly targeted at the older child. To describe the clinical profile of sick young infants presenting to a hospital and to define important signs and symptoms that will enable health workers to detect young infants with severe illness requiring hospital admission. Young infants aged 0-59 days presenting to a paediatric out-patient clinic were evaluated by a nurse using a standardised list of signs and symptoms. A paediatrician independently evaluated these children and decided whether they needed hospitalisation. A total of 685 young infants were enrolled, 22% of whom were <7 days of age. The commonest reasons for seeking care were jaundice in the 0-6-day group, skin problems in the 7-27-day group and cough in the 28-59-day group. The primary clinical diagnoses for admissions were sepsis in the 0-6- and 7-27-day groups and pneumonia in the 28-59-day group. Clinical signs and symptoms predicting severe illness requiring admission were general (history of fever, difficult feeding, not feeding well and temperature >37.5 degrees C) and respiratory (respiratory rate > or =60/min, severe chest in-drawing). General and respiratory signs are important predictors for severe illness in young infants. Training peripheral health workers to recognise these signs and to refer to hospital for further assessment and management might have a significant impact on young infant mortality.


Free R value: a novel statistical quantity for assessing the accuracy of crystal structures.
The determination of macromolecular structure by crystallography involves fitting atomic models to the observed diffraction data. The traditional measure of the quality of this fit, and presumably the accuracy of the model, is the R value. Despite stereochemical restraints, it is possible to overfit or 'misfit' the diffraction data: an incorrect model can be refined to fairly good R values as several recent examples have shown. Here I propose a reliable and unbiased indicator of the accuracy of such models. By analogy with the cross-validation method of testing statistical models I define a statistical quantity (R(free) (T) that measures the agreement between observed and computed structure factor amplitudes for a 'test' set of reflections that is omitted in the modelling and refinement process. As examples show, there is a high correlation between R(free) (T) and the accuracy of the atomic model phases. This is useful because experimental phase information is usually inaccurate, incomplete or unavailable. I expect that R(free) (T) will provide a measure of the information content of recently proposed models of thermal motion and disorder, time-averaging and bulk solvent.


Establishment of early diagnosis models for cervical precancerous lesions using large-scale cervical cancer screening datasets.
Human papilloma virus (HPV) DNA test was applied in cervical cancer screening as an effective cancer prevention strategy. The viral load of HPV generated by different assays attracted increasing attention on its potential value in disease diagnosis and progression discovery. In this study, three HPV testing datasets were assessed and compared, including Hybrid Capture 2 (n = 31,954), Aptima HPV E6E7 (n = 3269) and HPV Cobas 4800 (n = 13,342). Logistic regression models for diagnosing early cervical lesions of the three datasets were established and compared. The best variable factor combination (VL + BV) and dataset (HC2) were used for the establishment of six machine learning models. Models were evaluated and compared, and the best-performed model was validated. Our results show that viral load value was significantly correlated with cervical lesion stages in all three data sets. Viral Load and Bacterial Vaginosis were the best variable factor combination for logistic regression model establishment, and models based on the HC2 dataset performed best compared with the other two datasets. Machine learning method Xgboost generated the highest AUC value of models, which were 0.915, 0.9529, 0.9557, 0.9614 for diagnosing ASCUS higher, ASC-H higher, LSIL higher, and HSIL higher staged cervical lesions, indicating the acceptable accuracy of the selected diagnostic model. Our study demonstrates that HPV viral load and BV status were significantly associated with the early stages of cervical lesions. The best-performed models can serve as a useful tool to help diagnose cervical lesions early.


Yolk sac tumor identified at autopsy after surgical excision of immature sacrococcygeal teratoma.
Although the predictive value of immature elements in sacrococcygeal teratomas in unclear, there are reports of malignant recurrence after surgical resection of immature sacrococcygeal teratomas. The recurrent tumors are presumed to arise from small residual malignant foci not identified at the time of surgical resection. In this report a premature female infant was delivered at 29 week's gestation with a large sacrococcygeal teratoma. The tumor weighed 1,350 g. It was largely cystic with a focal nodular and variegated appearance. Histologically, the tumor was a grade 1 immature teratoma with a predominance of neuroglial elements. No malignant elements were identified in any of 26 sections examined. The infant died intraoperatively of cardiovascular complications related to the large vascular supply of the tumor but had a grossly complete resection of tumor. At autopsy, a small microscopic focus of yolk sac tumor was identified adjacent to the sacrum anteriorly. Had the infant survived, this focus might well have been a source for malignant recurrence.


An efficient procedure to stably introduce genes into an economically important pulp tree (Eucalyptus grandis x Eucalyptus urophylla).
Regeneration problems are one of the main limitations preventing the wider application of genetic engineering strategies to the genus Eucalyptus. Seedlings from Eucalyptus grandis x Eucalyptus urophylla were selected according to their regeneration (adventitious organogenesis) and transformation capacity. After in vitro cloning, the best genotype of 250 tested was transformed via Agrobacterium tumefaciens. A cinnamyl alcohol dehydrogenase (CAD) antisense cDNA from Eucalyptus gunnii was transferred, under the control of the 35S CaMV promoter with a double enhancer sequence, into a selected genotype. According to kanamycin resistance and PCR verification, 120 transformants were generated. 58% were significantly inhibited for CAD activity, and nine exhibited the highest down-regulation, ranging from 69 to 78% (22% residual activity). Southern blot hybridisation showed a low transgene copy number, ranging from 1 to 4, depending on the transgenic line. Northern analyses on the 5-16 and 3-23 lines (respectively one and two insertion sites) demonstrated the antisense origin of CAD gene inhibition. With respectively 26 and 22% of residual CAD activity, these two lines were considered as the most interesting and transferred to the greenhouse for further analyses.


Co-located Parent Coaching Services Within Pediatric Primary Care: Feasibility and Acceptability.
This project assessed the feasibility and acceptability of Parent Connext, a positive parenting program that integrates screening and co-located parent coaching within pediatric primary care. Eleven practices implemented Parent Connext in phases between November 1, 2016, and July 31, 2019. Screening and surveillance for parenting and family psychosocial concerns were performed during patient visits. Providers responded with brief motivational interviewing and referral. Parenting Specialists provided individualized parent coaching to referred caregivers. Screens were completed at 13,346 (65%) targeted visits, with 26% positive for concerns. Parent coaching was provided to 1,301 of 2,711 (48%) referred families (average 2.2 sessions per family). Providers and staff felt significantly more supported, confident, and knowledgeable about addressing parenting and family psychosocial concerns after implementing Parent Connext and felt the program improved their relationships with families and quality of care. Co-located parent coaching was found to be a feasible and worthwhile addition to pediatric primary care.


A CD40 Signalosome anchored in lipid rafts leads to constitutive activation of NF-kappaB and autonomous cell growth in B cell lymphomas.
B cell lineage non-Hodgkin's lymphomas (NHL-B) are neoplastic B cells that show dysregulated B lymphocyte growth characteristics. Unlike normal B cells, aggressive NHL-B cells show constitutive expression of nuclear NF-kappaB by maintaining an assembled, scaffold-like signaling platform, called a Signalosome within the lipid raft microdomain, extending from the cell membrane. The CD40 Signalosome appears to be initiated through autochthonous production and cognate binding of CD154 (CD40L, gp39) to CD40 by the lymphoma cell. Constitutive expression of NF-kappaB in NHL-B can be downregulated by treatment with antibodies to CD40 or CD154 that disrupt Signalosomes, inhibit lymphoma cell growth, and induce cell death. CD40 Signalosomes may provide a potentially vulnerable target for therapeutic intervention in NHL-B cells.


Design, synthesis, AML activity and molecular modeling of novel IDH2 inhibitors.
Enasidenib (AG-221) is the only approved IDH2 inhibitor, clinical study found Enasidenib have some side-effects. In this work, we synthesized series of novel s-triazine derivatives, and the in vitro and in vivo activity of anti-AML has been studied using AM7577 model. The cell activity found Ta and Th showed excellent inhibition to AM7577. We further used the HuKemia Acute Leukemia xenograft model to investigate the in vivo efficacy of compounds Ta and Th, compared with AG-221, although Ta and Th can't reduce the 2-HG level obviously, those two compounds can prolong the survival of rats. The research can expand the structure of novel IDH2 inhibitors and provide useful information for further research of novel AML drugs.


Fasting blood soluble RAGE may be causally implicated in impaired glucose metabolism in Chinese patients with primary hypertension.
We prepared to investigate the association of four well-defined polymorphisms in the receptor for advanced glycation end-products (RAGE) gene with the changes of fasting blood soluble RAGE (sRAGE) and endogenous secretory RAGE (esRAGE) and the risk for impaired glucose metabolism (IGM) in 1704 patients with primary hypertension, aiming to infer possible causality between sRAGE/esRAGE and IGM. This was a hospital-based case-control study, including 848 patients coexisting with IGM (the case group) and 856 patients with normal glucose tolerance (the control group). Fasting blood sRAGE and esRAGE concentrations were measured in 300 cases and 300 controls. There were significant differences in the genotypes/alleles of T-429C (rs1800625) and T-374A (rs1800624) polymorphisms between the case and control groups after Bonferroni correction (P<0.05/8). Adjusted estimates of above two polymorphisms for IGM risk were remarkably significant, especially under the recessive model (odd ratio [OR], 95% confidence interval [CI], P: 3.57, 1.95-5.18, 0.002 for T-429C and 3.49, 1.42-8.58, 0.007 for T-374A). Mean sRAGE and esRAGE concentrations were significantly higher in controls than in cases (P<0.001). Participants with the rs1800625 -429CC and -429TC genotypes had significantly lower sRAGE (417.3 and 473.6 vs. 502.3pg/mL, P<0.01) and esRAGE (230.1 and 298.0 vs. 340.4pg/mL, P<0.05) concentrations than those with the -429TT genotype in primary hypertensive patients with IGM. Further Mendelian randomization analysis revealed that per 100pg/mL reduction in fasting blood sRAGE and esRAGE was causally associated with 2.40-fold (95% CI: 1.46-3.94) and 2.65-fold (95% CI: 1.24-5.13) increased IGM risk, respectively. Our findings collectively demonstrate that fasting blood sRAGE and esRAGE may be causally implicated in IGM in primary hypertensive patients.


Local anesthetics in dentistry: clinical considerations, drug interactions, and novel formulations.
The pain control afforded by local anesthetics is the mainstay of dental practice. It is the medicolegal responsibility of all practicing dentists to possess a high level of expertise on the clinical use of these drugs. This article will discuss their clinical selection, recommended dosages, and new research developments that impact on dental medicine.


Screening on platelet LncRNA expression profile discloses novel residual platelet reactivity biomarker.
Double antiplatelet therapy (DAPT) has wide inter-individual variabilities in coronary heart disease (CHD) patients' responses, which undermines the prognosis effect in clinical practice. Long noncoding RNAs (lncRNAs) have been reported to be widely existed in platelets, albeit their potential roles in platelet responses to DAPT largely remains in the realm of the unknown. This study aims to screen differentially expressed lncRNAs responsible for high residual platelet reactivities under DAPT. We enrolled 144 CHD patients that received DAPT and assigned them to high platelet reactivity (HPR) group and baseline group according to their residual platelet reactivities. Statistical analysis and a series of experiments including microarray analysis, platelet reactivity screening, RNA isolation and lncRNA microarray analysis were explored to the research. We detected a total of 22,424 kinds of co-expressed lncRNAs in three pairs of patients between the HPR and baseline groups. We identified twenty differentially expressed lncRNAs and successfully validated three of them in the 144 patients. Ultimately, the expression of ensemble transcript ENST00000433442 was demonstrated as significantly correlated with high platelet reactivity (OR = 0.487, P = .035) by logistic regression analysis. There is a considerable amount of lncRNAs in platelets, and the downregulated expression of ENST00000433442 is an independent risk factor for high residual platelet reactivity in CHD patients under DAPT.


[Entrapment syndromes of the intercostal nerve and its branches].
Musculoskeletal chest pain poses a broad differential diagnosis, among which intercostal nerve involvement stands out. Its entrapment or that of any of its branches can resemble visceral pain and therefore can easily go unnoticed. With a good examination and the use of dynamic ultrasound, the diagnostic approach can be simpler. We present a 40-year-old man evaluated in a rehabilitation department for right lower rib pain, triggered by certain movements and associated with a pectus excavatum type thoracic deformity. Using dynamic ultrasound maneuvers, he was diagnosed with neuralgia of the 7th right intercostal nerve secondary to dynamic entrapment in the context of a thoracic deformity with costal hypermobility. We describe the clinical presentation, ultrasound imaging, treatment, and evolution after treatment. In this case, we describe entrapment syndromes of the intercostal nerve and its branches, their clinical and ultrasound diagnosis, and their therapeutic approach.


Development and validation of an analytical method for the determination of semicarbazide in fresh egg and in egg powder based on the use of liquid chromatography tandem mass spectrometry.
In this work the development of a method to determine semicarbazide (SEM) in fresh whole egg and in whole egg and egg white powders as those used in the food industry, by using LC-MS/MS is described. The method is based on a recently validated one for the determination of SEM in baby food. SEM is extracted from egg with 0.2 mol L(-1) hydrochloric acid and derivatised with 2-nitrobenzaldehyde. The extract is neutralised and purified by passing through a solid phase extraction (SPE) cartridge. The 2-nitrobenzaldehyde derivative is eluted with ethyl acetate. The eluate is evaporated to dryness and the residue re-dissolved in methanol:water mobile phase. SEM is determined by reversed-phase LC-MS/MS. The "in-house" validation of this method has been performed taking into account the "Harmonized guidelines for single-laboratory validation of methods of analysis" (IUPAC Technical report) and the Commission Decision (2002/657/EC). The performance characteristics of the method were established by in-house validation procedures employing assays with standard solutions, sample blanks and spiked samples. Linearity, matrix effect, trueness, precision, selectivity, limits of detection and of quantification were determined. The fitness for purpose of this method was assessed based on its performance characteristics. LOD's of 0.15 and 0.4 microg kg(-1) for whole fresh egg and industrial egg powders, respectively, were obtained. The LOQ for fresh whole egg was 0.2 microg kg(-1) and for industrial egg powders 0.8 microg kg(-1). Linear calibration curves were obtained in the ranges 1-100 and 0.8-400 microg kg(-1) for fresh whole egg and the egg powders, respectively.


Human string-pulling with and without a string: movement, sensory control, and memory.
String-pulling is a behavior that is allied to many daily acts and is an easily performed action featuring hand-over-hand movements to reel in a string (or rope). String-pulling has been used as a test of perceptual and cognitive functions in many animal species, including human children, but its movements and sensory control have not been characterized. Male and female university students (n = 68) performed target-based or memory-based string-pulling in which they pulled down a string suspended on an overhead pulley and immediately afterwards attempted to make the same movement in a memory-based test. Frame-by-frame video scoring was used to describe movements, eye-tracking and visual occluding glasses were used to assess sensory control, and a Matlab video-analysis procedure was used to describe kinematics. The string was advanced using five arm/hand movements: with lift and advance comprising fast up movements, and grasp, pull and push comprising slow down movements. Fingers closed 5 (pinky) through 1 (thumb) to make a whole-hand grasp and release in target-based string pulling but moved in a reverse sequence for the memory-based task. Target-based string pulling was not visually guided unless participants were instructed to grasp at a cue, and then vision featured eye-tracking of the target and pupil dilation with the grasp, but there was no relation between eye events for memory-based string-pulling. For target-based string-pulling the left and right hands advanced the string with both independent and concurrent movement but only independent movements were featured in a more symmetrical memory-based movement. The results are discussed in relation to the sensory control of hand movements, contemporary theories of the neural control of hand movements, and species differences in string-pulling.


Polygenic risk for schizophrenia and subcortical brain anatomy in the UK Biobank cohort.
Research has shown differences in subcortical brain volumes between participants with schizophrenia and healthy controls. However, none of these differences have been found to associate with schizophrenia polygenic risk. Here, in a large sample (n = 14,701) of unaffected participants from the UK Biobank, we test whether schizophrenia polygenic risk scores (PRS) limited to specific gene-sets predict subcortical brain volumes. We compare associations with schizophrenia PRS at the whole genome level ('genomic', including all SNPs associated with the disorder at a p-value threshold < 0.05) with 'genic' PRS (based on SNPs in the vicinity of known genes), 'intergenic' PRS (based on the remaining SNPs), and genic PRS limited to SNPs within 7 gene-sets previously found to be enriched for genetic association with schizophrenia ('abnormal behaviour,' 'abnormal long-term potentiation,' 'abnormal nervous system electrophysiology,' 'FMRP targets,' '5HT2C channels,' 'CaV2 channels' and 'loss-of-function intolerant genes'). We observe a negative association between the 'abnormal behaviour' gene-set PRS and volume of the right thalamus that survived correction for multiple testing (ß = -0.031, pFDR = 0.005) and was robust to different schizophrenia PRS p-value thresholds. In contrast, the only association with genomic PRS surviving correction for multiple testing was for right pallidum, which was observed using a schizophrenia PRS p-value threshold < 0.01 (ß = -0.032, p = 0.0003, pFDR = 0.02), but not when using other PRS P-value thresholds. We conclude that schizophrenia PRS limited to functional gene sets may provide a better means of capturing differences in subcortical brain volume than whole genome PRS approaches.


Early unilateral cochlear implantation promotes mature cortical asymmetries in adolescents who are deaf.
Unilateral cochlear implant (CI) stimulation establishes hearing to children who are deaf but compromises bilateral auditory development if a second implant is not provided within ∼ 1.5 years. In this study we asked: 1) What are the cortical consequences of missing this early sensitive period once children reach adolescence? 2) What are the effects of unilateral deprivation on the pathways from the opposite ear? Cortical responses were recorded from 64-cephalic electrodes within the first week of bilateral CI activation in 34 adolescents who had over 10 years of unilateral right CI experience and in 16 normal hearing peers. Cortical activation underlying the evoked peaks was localized to areas of the brain using beamformer imaging. The first CI evoked activity which was more strongly lateralized to the contralateral left hemisphere than normal, with abnormal recruitment of the left prefrontal cortex (involved in cognition/attention), left temporo-parietal-occipital junction (multi-modal integration), and right precuneus (visual processing) region. CI stimulation in the opposite deprived ear evoked atypical cortical responses with abnormally large and widespread dipole activity across the cortex. Thus, using a unilateral CI to hear beyond the period of cortical maturation causes lasting asymmetries in the auditory system, requires recruitment of additional cortical areas to support hearing, and does little to protect the unstimulated pathways from effects of auditory deprivation. The persistence of this reorganization into maturity could signal a closing of a sensitive period for promoting auditory development on the deprived side.


The relationship of depression treatment quality indicators to employee absenteeism.
Although employers commonly review administrative database indicators to assess depression treatment quality, they do not know whether these indicators predict relevant outcomes like absenteeism. In 230 employed patients in five health plans, we tested how administrative database-derived indicators for antidepressant medication and psychotherapy provided during the first 6 months of a new depression treatment episode predicted patient-reported absenteeism change over 12 months. The medication indicator was not significantly associated with absenteeism change over 12 months (p = .64); however, the psychotherapy indicator was significantly associated with an average 26.1% improvement in absenteeism over 12 months (p < .05). If subsequent studies confirm the results we report, quality monitoring initiatives interested in employer-relevant indicators of depression treatment quality should examine administrative database indicators of psychotherapy.


Sequence polymorphism of mitochondrial DNA control region in Japanese.
Sequence polymorphisms of the mitochondrial DNA (mtDNA) control region, hypervariable regions I and II, from 100 unrelated Japanese were determined by PCR amplification and direct sequencing. Sequences of 404 nucleotides for hypervariable region I and 379 nucleotides for region II were obtained. Variable sites (85 and 45) were revealed in region I and region II, respectively, as compared to the reference sequence, and a total of 96 different genetic patterns from both regions I and II were determined. A point mutation heteroplasmy was observed at the ratio of approximately 50:50 from one individual at the sequence position 151 showing a nucleotide transition from C to T. The probability of identity was estimated as 2.3% for region I, 3.9% for region II, and 1.1% combined for both regions. These results suggest that sequence polymorphism of mtDNA control region would be very useful in forensic practice as a marker for individual identification.


Herniation through the foramen of Morgagni: early diagnosis and treatment.
The majority of congenital diaphragmatic hernias (CDH) occur through the foramen of Bochdalek; herniation through the foramen of Morgagni (MH) is rare. Fifteen children (12 males and 3 females) with congenital MH (7 right, 3 left, 5 bilateral) were treated over a period of 15 years, comprising 11% of a total of 135 children with different types of CDH. The majority (12, 80%) had repeated chest infections. In 1 the hernia was discovered accidentally during evaluation of trauma, and another presented in the neonatal period with acute respiratory distress (ARD). The diagnosis was made on plain lateral chest radiograph when there was anterior herniation of bowel loops, and in these cases the diagnosis was confirmed by barium enema. Computed tomography was useful in preoperative diagnosis when the hernia contents were solid. All patients were operated upon transabdominally except 1, and in all cases there was a hernia sac. Associated anomalies were present in 10 (66.7%) patients: 4 (26.7%) had malrotation, 4 (26.7%) congenital heart disease, and 3 (20%) Down's syndrome. Our study shows a relatively high frequency of MH in our patients. MH rarely presents in the neonatal period, but when it does, it causes ARD. The majority of patients with MH present beyond the neonatal period with repeated attacks of chest infection, and although late-presenting MH is relatively benign, it nevertheless causes significant morbidity. Thus, clinical awareness and early diagnosis and surgical treatment are important factors.


Airway immunopathology of asthma with exercise-induced bronchoconstriction.
Exercise-induced bronchoconstriction (EIB) is a common cause of symptoms in a subgroup of asthmatic subjects. The pathobiology that makes this group of asthmatic subjects susceptible to bronchoconstriction after a brief period of exercise remains poorly understood. We sought to determine whether there are differences in lower airway inflammation and production of cytokines and eicosanoids between asthmatic subjects with and without EIB. Two distinct groups of asthmatic subjects based on a priori definitions were identified, one with moderate-to-severe EIB and the other without significant bronchoconstriction after exercise challenge. Both groups met the definition of asthma on the basis of bronchodilator response, bronchial hyperresponsiveness, or both. A comparative immunopathology study was conducted by using induced sputum to identify differences in lower airway inflammation and production of cytokines and eicosanoids. The groups had similar baseline lung function and bronchodilator response and did not have any asthma exacerbations within the prior year. The concentration of columnar epithelial cells was markedly higher in the group with EIB (1.4 x 10(5) vs 2.9 x 10(4) cells/mL, P=.01). The concentration of eosinophils was higher in the group with EIB (3.6 x 10(4) vs 4.9 x 10(3) cells/mL P=.04). Cysteinyl leukotrienes (CysLTs; 727.7 vs 151.9 pg/mL, P=.01) and the ratio of CysLTs to prostaglandin E(2) (1.85 vs 1.04, P=.002) in the airways were higher in the group with EIB. Injury to the airway epithelium, overexpression of CysLTs, relative under production of prostaglandin E(2), and greater airway eosinophilia are distinctive immunopathologic features of asthma with EIB.


Localisation of the temperature-sensitive defect in the nucleoprotein of an influenza A/FPV/Rostock/34 virus.
The nucleotide sequences of the nucleoprotein (NP) genes of fowl plague virus (FPV) and of a temperature-sensitive (ts) mutant (ts81) derived therefrom have been determined. The ts81-NP nucleotide sequence possesses a single nucleotide substitution in comparison to the wild type. This causes an amino acid exchange at position 332 of the NP. An alanine in the wild type-NP is substituted by a threonine in ts81-NP. This substitution leads to a significant difference in the secondary structure prediction. Although this mutation is located within the karyophilic region of the NP, the accumulation of the NP in ts81-infected cells is not significantly affected at 40 degrees C. Therefore, we assume that the cooperation with one of the polymerase proteins (P) is interfered with at 40 degrees C, leading to the loss of viral vRNA or replicative cRNA synthesis. The comparison of the FPV-NP nucleotide sequence to a previously published sequence of the same strain (Tomley and Roditi, 1984) highlights ten nucleotide differences, four of them leading to amino acid substitutions.


Multi-gene amplification: simultaneous detection of three virulence genes in diarrhoeal stool.
Enterotoxigenic Escherichia coli (ETEC) and Shigella account for a substantial proportion of acute diarrhoeal illnesses among Third-World children. Rapid detection of these infectious agents in faeces followed by the prompt implementation of public health measures could help reduce their spread during the early phase of epidemics. Towards this end, three pairs of synthetic oligonucleotide primers were prepared and shown to hybridize specifically to the genes encoding the heat-stable (ST) and the heat-labile (LT) enterotoxins of ETEC and to invasion-associated loci (ial) of the large Shigella virulence plasmid. When the three primer pairs were used together in the polymerase chain reaction (PCR), the three corresponding genetic loci could be simultaneously amplified using DNA extracted directly from stool; the amplified products were readily detected by ST-, LT- and ial-specific, alkaline phosphatase-labelled oligonucleotide probes (AP probes). The performance of this system was evaluated in a Mayan community in southeastern Mexico, where diarrhoeal illnesses are a common cause of childhood morbidity and mortality. Using only simple and inexpensive laboratory equipment, multigene amplification with these primers and probes led to the identification of ETEC and/or Shigella in the stools of 20 out of 71 children with diarrhoea; the procedure could be completed in seven hours and was more sensitive than conventional diagnostic tests or DNA probes used without amplification.


MICS-1 interacts with mitochondrial ATAD-3 and modulates lifespan in C. elegans.
Caenorhabditis elegans open reading frame T21C9.1 encodes an uncharacterized protein, which is here named MICS-1 (mitochondrial scaffolding protein-1). It is predicted to be the homolog of human outer mitochondrial membrane protein 25 (OMP25 or synaptojanin-2-binding protein), which is a PDZ domain containing protein with a putative role in cellular stress response pathways. Here, we provide evidence that MICS-1 is an interacting partner of mitochondrial protein ATAD-3 (homologue of human ATAD3), which is essential for C. elegans development. We demonstrate that mics-1(RNAi) animals or mics-1 mutants display enhanced longevity with an increased mean lifespan of up to 54% compared to control animals. Of note, also atad-3(RNAi) promoted longevity, although to a lesser extend (29% compared to controls). In addition, thermal stress of mics-1 mutants induced low reactive oxygen species (ROS) production, whereas atad-3(RNAi) animals were highly sensitive to this assay, displaying drastically increased ROS levels. Further studies revealed that MICS-1 and ATAD-3 associated longevity was partially dependent on the presence of DAF-16. However, for both conditions, we also found a DAF-16 independent extension of lifespan. Finally, we observed an additional lifespan extension in mics-1 mutants when subjected to atad-3(RNAi) whereas heat induced ROS production was even aggravated under this condition. This suggests (partially) independent effects of MICS-1 and ATAD-3 on lifespan and ROS production in vivo.


The Effectiveness of Cigarette Pack Health Warning Labels with Religious Messages in an Urban Setting in Indonesia: A Cross-Sectional Study.
This study sought to assess the effectiveness of religious cigarette health warning labels (HWLs) in Indonesia, a country with a high public health burden from tobacco use. The study tested different religious and nonreligious messages related to suicide, secondhand smoke (SHS) and gangrene. Participants were smokers and non-smokers from Surabaya, Indonesia (n = 817). Participants rated each HWL for its effectiveness on a scale of 1 to 10 (1 = "not at all", 10 = "extremely") with respect to 10 items. Nonreligious HWLs were marginally superior for SHS and suicide while religious HWLs were marginally superior for gangrene. Given the close rating scores between religious and nonreligious HWLs, they were functionally equal in effectiveness. With proper assessment of potential unintended consequences, the implementation of religious HWLs could be considered for a proportion of HWLs.


Nuclear accumulation of Yes-Associated Protein (YAP) maintains the survival of doxorubicin-induced senescent cells by promoting survivin expression.
Although chemotherapeutic drugs can induce senescence to prohibit further division of tumor cells, senescence could also promote tumorigenesis mainly through a senescence-associated secretory phenotype. Therefore, senescent tumor cells should be eliminated immediately to prevent drug resistance and recurrence. Here, we used a doxorubicin-induced senescence model to explore the mechanism underlying the survival of therapy-induced senescent cells. After low-dose doxorubicin treatment, tumor cells turned on a senescence program and became large and flattened, increasing their contact area with the extracellular matrix (ECM). Furthermore, Yes-associated protein (YAP) accumulated in the nucleus and YAP activity was increased in doxorubicin-induced senescent cells. Knockdown of YAP increased the sensitivity of cells to low-dose doxorubicin treatment, causing apoptosis rather than senescence. Moreover, the anti-apoptotic gene survivin, a YAP target gene, was overexpressed in senescent cells. Inhibition of survivin could lead to selective elimination of senescent cells through apoptosis. Our study indicates that nuclear accumulation of YAP could promote the survival of senescent cells by increasing survivin expression. Therefore, targeting YAP or survivin might be a new strategy for clearing senescent cancer cells during drug treatment.


Persistence of a large population of exhausted monoclonal B cells in mixed cryoglobuliemia after the eradication of hepatitis C virus infection.
Functionally exhausted and mostly autoreactive B-cells with a peculiar CD21(low)CD11c(+) phenotype accumulate in several human immunological disorders including common variable immunodeficiency, HIV infection and rheumatoid arthritis. In HCV-associated mixed cryoglobulinemia (MC) there is accumulation of exhausted clonal B cells expressing a V(H)1-69-encoded cross-reactive idiotype; these cells are phenotypically heterogeneous, displaying either a CD21(low)CD11c(+) or a marginal zone (MZ)-like (IgM(+)CD27(+)CD21(+)CD11c(-)) phenotype. Irrespective of their phenotype, V(H)1-69(+) B-cells are unresponsive to the stimulation of Toll-like receptor 9 (TLR9). We investigated the fate of these cells after the eradication of HCV. Fourteen MC patients were studied before and after antiviral therapy. V(H)1-69(+) B-cells were identified using the G6 monoclonal antibody and their phenotype and responsiveness to the stimulation of TLR9 were investigated. In seven virological non-responders, cryoglobulin levels and the number and phenotype of V(H)1-69(+) B cells remained substantially unchanged. By contrast, in sustained viral responders cryoglobulinemia subsided and the number of V(H)1-69(+) B cells declined. However, high proportions of MZ-like V(H)1-69(+) B cells retaining unresponsiveness to TLR9 stimulation persisted for several months in these patients. Clonal expansion of CD21(low) V(H)1-69(+) B cells may depend on continual stimulation by HCV, whereas their MZ-like counterparts may persist for years after the eradication of infection. Prolonged survival of exhausted MZ-like B cells after withdrawal of the initial inciting stimulus may contribute to the accumulation of autoreactive B cells in immunological disorders.


Left ventricular systolic function in Nigerian children infected with HIV/AIDS: a cross-sectional study.
Cardiac complications contribute significantly to morbidity and mortality in children with HIV/AIDS. These rates have been under-reported in sub-Saharan African children. This was an observational, cross-sectional Doppler echocardiographic study of ventricular systolic function, performed at a tertiary clinic on children with HIV/AIDS. Left ventricular systolic dysfunction was present in 27.0% of the children with HIV infection and 81.2% of those with AIDS. The mean fractional shortening in the AIDS group (31.6 ± 9.5%) was significantly lower than in the HIV-infected group (35.3 ± 10.5%, p = 0.001). A significant correlation was found with CD4(+) cell count and age, and these were the best predictors of left ventricular systolic dysfunction in the stepwise multiple regression analysis (r = 0.396, p = 0.038; r = -0.212, p = 0.025, respectively). Left ventricular systolic dysfunction is common in Nigerian children with HIV/AIDS.


Antitumor activity of the aqueous-alcoholic extracts from unripe cotton ball of Gossypium indicum.
The present study investigated the antitumor activity of the aqueous-alcoholic extracts from unripe cotton balls of Gossypium indicum. An Exposure of murine B16 melanoma and L1210 lymphoma cells to the extracts resulted in their severe deaths in time- and concentration-dependent manners. Of the extracts, hydrophilic fractions were most efficacious for the antitumor activity and found to contain certain amounts of catechin and its derivatives. The hydrophilic extract fraction C36B2-8 had approximately 10 times more cytotoxic effects on B12 and L1210 cells than on isolated murine thymocytes. High concentrations (> 150 micrograms/ml) of C 36B3-8 mainly induced necrotic cell death. At low concentrations (< 100 micrograms/ml), however, C 36B3-8 induced not only necrosis but also apoptosis of the two tumor cell lines, which was proved by the TUNEL staining and DNA fragmentation techniques. The data indicate that certain ingredients of the cotton ball extract of G. indicum have an antitumor activity.