Alterations of G1-S checkpoint in chordoma: the prognostic impact of p53 overexpression.
To the authors' knowledge, little is known regarding the alterations of G(1)-S checkpoint and their significance in chordoma, a rare bone tumor. The authors investigated the clinicopathologic relevance of cell cycle abnormalities in chordoma. The expression levels of p53, murine double minute 2 (MDM2), retinoblastoma protein (pRb), cyclin D1, p16(INK4a), and p27(Kip1) were investigated using immunohistochemical techniques; p53 mutations were studied by polymerase chain reaction (PCR)-single-strand conformation polymorphism, and mdm2 amplification was analyzed using real-time quantitative PCR. The results were compared with clinicopathologic parameters in 101 lesions. Approximately 10-45% of primary tumors presented alterations of p53, MDM2, cyclin D1, and pRb proteins; most tumors lacked expression of p16(INK4a) and p27(Kip1). Alterations of p53, MDM2, cyclin D1, and pRb proteins were found to have cooperative effects on both higher proliferative ability (MIB-1 labeling index [LI]) and increased nuclear pleomorphism, a previously described prognostic indicator for patients with chordoma. Multivariate analyses revealed that, among these alterations, p53 overexpression was the only independent factor for higher MIB-1 LI. At the genetic level, mdm2 gene amplification was detected in 15.4% of the lesions but did not correlate with MDM2 overexpression or other clinicopathologic parameters. No p53 mutations were detected in the current series. Survival analysis revealed that p53 overexpression, but no other cell cycle alterations, was associated with a reduced overall survival. Accumulation of cell cycle alterations led to an increased MIB-1 LI and nuclear pleomorphism, a previously described prognostic indicator in chordoma. The authors believe that p53 overexpression in particular is associated with an unfavorable prognosis in patients with chordoma.


The multiplexed CRISPR targeting platforms.
The discovery and engineering of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) in the past several years have revolutionized biomedical research. The CRISPR technology showed great potential to advance detection, prevention, and treatment of human diseases in the near future. Compared to previous developed genome editing approaches, such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), the CRISPR-based systems have numerous advantages. One example is that the CRISPR systems can be easily adopted to efficiently target multiple genes simultaneously. Several strategies and toolboxes have been developed to achieve multiplexed targeting using the CRISPR systems. In this short review, we will discuss the principle, approach, and application of these strategies.


Structural Determinants of Redox Conduction Favor Robustness over Tunability in Microbial Cytochrome Nanowires.
Structural determinants of a 103-fold variation in electrical conductivity for helical homopolymers of tetra-, hexa-, and octa-heme cytochromes (named Omc- E, S, and Z, respectively) from Geobacter sulfurreducens are investigated with the Pathways model for electron tunneling, classical molecular dynamics, and hybrid quantum/classical molecular mechanics. Thermally averaged electronic couplings for through-space heme-to-heme electron transfer in the "nanowires" computed with density functional theory are ≤0.015 eV. Pathways analyses also indicate that couplings match within a factor of 5 for all "nanowires", but some alternative tunneling routes are found involving covalent protein backbone bonds (Omc- S and Z) or propionic acid-ligating His H-bonds on adjacent hemes (OmcZ). Reorganization energies computed from electrostatic vertical energy gaps or a version of the Marcus continuum expression parameterized on the total (donor + acceptor) solvent-accessible surface area typically agree within 20% and fall within the range 0.48-0.98 eV. Reaction free energies in all three "nanowires" are ≤|0.28| eV, even though Coulombic interactions primarily tune the site redox energies by 0.7-1.2 eV. Given the conserved energetic parameters, redox conductivity differs by < 103-fold among the cytochrome "nanowires". Redox currents do not exceed 3.0 × 10-3 pA at a physiologically relevant 0.1 V bias, with the slowest electron transfers being on a (μs) timescale much faster than typical (ms) enzymatic turnovers. Thus, the "nanowires" are proposed to be functionally robust to variations in structure that provide a habitat-customized protein interface. The 30 pA to 30 nA variation in conductivity previously reported from atomic force microscopy experiments is not intrinsic to the structures and/or does not result from the physiologically relevant redox conduction mechanism.


A liposome-based energy conversion system for accelerating the multi-enzyme reactions.
We report the first example of a liposome-based energy conversion system that is useful for entrapping enzymes and NAD coenzyme to accelerate multi-step enzymatic reactions. The liposome generates a much higher catalytic current compared with the non-liposome system, which is in good consistency with numerical simulations.


Vulnerabilities Associated with Post-disaster Declines in HIV-testing: Decomposing the Impact of Hurricane Sandy.
Using Interrupted Time Series Analysis and generalized estimating equations, this study identifies factors that influence the size and significance of Hurricane Sandy's estimated impact on HIV testing in 90 core-based statistical areas from January 1, 2011 to December 31, 2013. Generalized estimating equations were used to examine the effects of sociodemographic and storm-related variables on relative change in HIV testing resulting from Interrupted Time Series analyses. There is a significant negative relationship between HIV prevalence and the relative change in testing at all time periods. A one unit increase in HIV prevalence corresponds to a 35% decrease in relative testing the week of the storm and a 14% decrease in relative testing at week twelve. Building loss was also negatively associated with relative change for all time points. For example, a one unit increase in building loss at week 0 corresponds with an 8% decrease in the relative change in testing (p=0.0001) and a 2% at week twelve (p=0.001). Our results demonstrate that HIV testing can be negatively affected during public health emergencies. Communities with high percentages of building loss and significant HIV disease burden should prioritize resumption of testing to support HIV prevention.


Nocardia kirovani cell walls possess a lipidic fraction, a lipolysaccharidic fraction and a peptidoglycan. The lipidic part is composed of glycerides, C(16) and C(18) free fatty acids, nocardic acids and of a carotenoid pigment. The lipopolysaccharidic part is composed of an arabinogalactan esterified by nocardic acids. The glycan strand consists of alternating 1-->4 linked N-acetylglucosamine and N-glycolylmuramic acid residues substituted by a peptide subunit in which the aminoacids Ala, Glu, meso-Dap are present in the molar proportions 1,7:1,2:1.


Ultraviolet absorbance titration for determining stability constants of humic substances with Cu(II) and Hg(II).
We describe an ultraviolet (UV) absorbance titration method that can be used to determine complexing capacities (C(L)) and conditional stability constants (logK) of humic substances (HSs) with metal ions such as Cu(II) and Hg(II). Two fulvic acids (FA) and one humic acid (HA) were used for this study. UV absorbance of HSs gradually increased with the addition of Cu(II) or Hg(II) after blank correction, and these increases followed the theoretical 1:1 (ligand:metal ion) binding model. The results from the absorbance titration calculation for HSs with Cu(II) and Hg(II) compared well with those from fluorescence quenching titration. The titration of the model compound l-tyrosine with Cu(II) proved the validity of this method, and the K and C(L) were within 2.3% and 7.4% of the fluorescence quenching titration. The results suggest that the UV absorbance titration can be used to study the binding capacities of HSs and/or dissolved organic matter (DOM) with trace metals. The advantages and disadvantages of the absorbance titration method were also discussed.


L-Lactic acid production from raw cassava starch in a circulating loop bioreactor with cells immobilized in loofa (Luffa cylindrica).
L-Lactic acid was produced from raw cassava starch, by simultaneous enzyme production, starch saccharification and fermentation in a circulating loop bioreactor with Aspergillus awamori and Lactococcus lactis spp. lactis immobilized in loofa sponge. A. awamori was immobilized directly in cylindrical loofa sponge while the L. lactis was immobilized in a loofa sponge alginate gel cube. In the loofa sponge alginate gel cube, the sponge serves as skeletal support for the gel with the cells. The alginate gel formed a hard outer layer covering the soft porous gel inside. By controlling the rate and frequency of broth circulation between the riser and downcomer columns, the riser could be maintained under aerobic condition while the downcomer was under anaerobic condition. Repeated fed-batch L-lactic acid production was performed for more than 400 h and the average lactic acid yield and productivity from raw cassava starch were 0.76 g lactic acid g(-1) starch and 1.6 g lactic acid l(-1) h(-1), respectively.


Effect of l-Arginine Supplementation on Blood Pressure in Adults: A Systematic Review and Dose-Response Meta-analysis of Randomized Clinical Trials.
A decade-old meta-analytic work indicated that l-arginine supplementation might have a blood pressure (BP)-lowering effect in different populations. However, several relevant investigations have emerged in the last 10 y, and an up-to-date systematic review and meta-analysis on this topic is currently lacking. Therefore, we aimed to examine the impact of l-arginine supplementation on BP by conducting a systematic review and dose-response meta-analysis of randomized placebo-controlled clinical trials (RCTs). We searched online databases using relevant keywords up to April 2021 to identify RCTs using oral l-arginine on systolic BP (SBP) and diastolic BP (DBP) in adults. Inclusion criteria were adult participants and an intervention duration ≥4 d. Exclusion criteria were the use of l-arginine infusion and acute interventions. A random-effects model was used to estimate the weighted mean difference (WMD) and 95% CI. Twenty-two RCTs with 30 effect sizes were included in this meta-analysis. The pooled analysis demonstrated significant decreases in SBP (WMD = -6.40 mmHg; 95% CI: -8.74, -4.05; P < 0.001) and DBP (WMD = -2.64 mmHg; 95% CI: -3.94, -1.40; P < 0.001) after l-arginine supplementation. Subgroup analysis showed significant reductions in SBP and DBP regardless of baseline BP category (normotensive, hypertensive), study duration (≤24 d, >24 d), sex (female, male), health status (healthy, unhealthy), and BMI (normal, overweight, obese). No significant changes were observed with dosages >9 g/d, trial duration >24 d, or in obese individuals. l-Arginine supplementation also appears to decrease DBP more effectively in females than in males. Moreover, meta-regression analysis for DBP demonstrated a significant relation between the dose of l-arginine intake and changes in DBP (P = 0.020). In the nonlinear dose-response analysis, the effective dosage of l-arginine supplementation was detected to be ≥4 g/d for SBP (P = 0.034), independent of trial duration. Overall, l-arginine supplementation may be effective for decreasing BP. This study was registered at PROSPERO as CRD42021242772.


The arterialized saphenous venous flow-through flap with dual venous drainage.
Venous flow-through flaps are well-described options for small defects where donor site morbidity is undesirable or in areas where useful local veins are in close proximity to the defect, particularly in the extremities. However, higher rates of flap loss have limited their utility. The saphenous venous flap in particular has been widely sought as a useful flap, and while arterialization of this flap improved survival rates, congestion has remained a limiting feature. We describe report a modification in the design of saphenous venous flaps, whereby an arterialized flap is provided with a separate source of venous drainage, and demonstrate survival of substantially larger venous flaps than previously reported. In five consecutive patients, we describe three main modifications to the saphenous venous flap as previously described: (a) Using arterialized flaps only; (b) Reversing the flap to allow unimpeded flow during arterialization; and (c) Anastomosing additional vein(s) that are not connected to the central vein-especially at the periphery of the flap for true venous drainage. There was a 0% complete flap loss rate (with only one case of superficial partial loss), and ultimately better survival than previous series of saphenous venous flaps described to date. The success of these techniques offers the potential to re-establish flow to large segmental losses to axial arteries, offer safe and definitive flap coverage to traumatic wounds, improve the array of flap options in this setting, and minimize donor site morbidity.


Mechanosensitive components of integrin adhesions: Role of vinculin.
External forces play a key role in shaping development and normal physiology. Aberrant responses to forces, or changes in the nature of such forces, are implicated in a variety of diseases. Cells contain several types of adhesions, linking them to their external environment. It is through these adhesions that forces are both sensed (from the outside inwards) and applied (from inside to out). Furthermore, several adhesion-based proteins are sensitive to changes in intracellular forces, utilising them for activation and regulation. Here, we outline how vinculin, a key component of integrin-mediated adhesions linking the actin cytoskeleton to the extracellular matrix (ECM), is regulated by force and acts as force transducing protein. We discuss the role of vinculin in vivo and its place in health and disease; summarise the proposed mechanisms by which vinculin is recruited to and activated at integrin-ECM adhesions; and discuss recent findings that place vinculin as the major force sensing and transmitting component of cell-matrix adhesion complexes. Finally, we discuss the role of vinculin in regulating the cellular responses to both the physical properties of the external environment and to externally applied physical stimuli.


Modification of the association between maternal smoke exposure and congenital heart defects by polymorphisms in glutathione S-transferase genes.
Congenital heart defects (CHDs) arise through various combinations of genetic and environmental factors. Our study explores how polymorphisms in the glutathione S-transferase (GST) genes affect the association between cigarette smoke exposure and CHDs. We analysed 299 mothers of children with CHDs and 284 mothers of children without any abnormalities who were recruited from six hospitals. The hair nicotine concentration (HNC) was used to quantify maternal smoke exposure, and the maternal GSTT1, and GSTM1 and GSTP1 genes were sequenced. We found a trend of higher adjusted odds ratios with higher maternal HNC levels, suggesting a dose-response relationship between maternal smoke exposure and CHDs. The lowest HNC range associated with an increased risk of CHDs was 0.213-0.319 ng/mg among the mothers with functional deletions of GSTM1 or GSTT1and 0.319-0.573 ng/mg among the mothers with normal copies of GSTM1 and GSTT1. In addition, the adjusted odds ratio for an HNC of >0.573 ng/mg was 38.53 among the mothers with the GSTP1 AG or GG genotype, which was 7.76 (χ(2) = 6.702, p = 0.010) times greater than the AOR in the mothers with GSTP1 AA genotype. Our study suggests that polymorphisms of maternal GST genes may modify the association of maternal smoke exposure with CHDs.


RTN1-C mediates cerebral ischemia/reperfusion injury via ER stress and mitochondria-associated apoptosis pathways.
The reticulon family has been found to induce apoptosis, inhibit axon regeneration and regulate protein trafficking. However, little is known about the mechanisms of how reticulon proteins are involved in neuronal death-promoting processes during ischemia. Here, we report that the expression of Reticulon Protein 1-C (RTN1-C) was associated with the progression of cerebral ischemia/reperfusion (I/R) injury. Using a combination of rat middle cerebral artery occlusion (MCAO) stroke and oxygen-glucose deprivation followed by reoxygenation (OGD/R) models, we determined that the expression of RTN1-C was significantly increased during cerebral ischemic/reperfusion. RTN1-C overexpression induced apoptosis and increased the cell vulnerability to ischemic injury, whereas RTN1-C knockdown reversed ischemia-induced apoptosis and attenuated the vulnerability of OGD/R-treated neural cells. Mechanistically, we demonstrated that RTN1-C mediated OGD/R-induced apoptosis through ER stress and mitochondria-associated pathways. RTN1-C interacted with Bcl-xL and increased its localization in the ER, thus reducing the anti-apoptotic activity of Bcl-xL. Most importantly, knockdown of Rtn1-c expression in vivo attenuated apoptosis in MCAO rats and reduced the extent of I/R-induced brain injury, as assessed by infarct volume and neurological score. Collectively, these data support for the first time that RTN1-C may represent a novel candidate for therapies against cerebral ischemia/reperfusion injury.


A comparative assessment of pomegranate extract, sodium hypochlorite, chlorhexidine, Myrrh (Commiphora molmol), tulsi extract against Enterococcus faecalis, Fusobacterium nucleatum and Staphylococci epidermidis.
The presence of pathogenic bacteria, toxins and byproducts in the root canal system show a decisive part in success of endodontic therapy. Thus, the complete removal of this bacterium is highly desirable. Several intracanal medicaments were tried to disinfect the root canal before obturation. The present study was conducted to compare Pomegranate, sodium hypochlorite, Chlorhexidine, Myrrh (Commiphora molmol), tulsi extract against Enterococcus faecalis, Fusobacterium nucleatum and Staphylococci epidermidis. Cross-sectional observational prospective study. Aqueous extract of 20% pomegranate peel, 20% pomegranate peel, 0.2% CHX, 2.5% sodium hypochlorite, Tulsi extract and Myrrh (Commiphora molmol) was used as agent against E. faecalis, F. nucleatum and Staphylococci Epidermidis. Zone of inhibition and minimum inhibitory concentration (MIC) was calculated and compared using analysis of variance and Mann-Whitney test. The information was statistically evaluated with SPSS software version 20 with P < 0.05. The mean zone of inhibition against E. faecalis, F. nucleatum and S. Epidermidis was highest in chlorhexidine and sodium hypochlorite groups compared to herbal groups. MIC was least with group III followed by group II against all bacterial species (P < 0.05). Sodium hypochlorite found comparatively better followed by chlorhexidine and other agents against E. faecalis, F nucleatum and S. epidermidis.


[A pediatric case of Alveolar Soft Part Sarcoma].
Alveolar Soft Part Sarcoma (ASPS) is a rare malignant tumor arising from skeletal muscles and occurring usually in young adults. It is a solid chemo resistant tumor. At time of diagnosis the disease is usually advanced and lung metastasis are present in 65% of the cases. The translocation (X; 17), producing an ASPL-TFE3 transcript fusion which is detected on tumoral cells wall, is specific of ASPS. The curative treatment is surgical. We report the case of a child with an ASPS of the tongue and lung metastasis. This report presents two particularities. Firstly this type of sarcoma is very exceptional in a 2-year-old child. Secondly ASPS is not usually susceptible to chemotherapy.


High survival rate of metaphase II human oocytes after first polar body biopsy and vitrification: determining the effect of previtrification conditions.
To use metaphase II (MII) bovine oocytes as a model for MII human oocyte cryopreservation and to determine the effect of different previtrification equilibration temperatures, vitrification solutions, zona slitting, and first polar body biopsy on in vitro and in vivo developmental competence of MII human oocytes after the CryoLoop vitrification method. In vitro and in vivo studies. A private infertility clinic. Women undergoing infertility treatment. Metaphase II stage bovine and MII human oocytes underwent first polar body biopsy before cryopreservation in different vitrification conditions, and human oocytes were fertilized by intracytoplasmic sperm injection after warming. The resulting embryos were transferred into women undergoing infertility treatment. Postvitrification morphologic survival, in vitro blastocyst development, and clinical outcome after ET. The equilibration temperature had a significant effect on cryosurvival of both bovine and human oocytes. High (97%-99%) postvitrification survival was achieved for both MII bovine and human oocytes, and high fertilization (90%-97%) at 35 degrees C to 37 degrees C, blastocyst development (18%-45%), and pregnancy (50%) rates were achieved at 35 degrees C with 5.0 mol/L ethylene glycol + 1.3 mol/L dimethyl sulfoxide for MII human oocytes that underwent first polar body biopsy. Previtrification equilibration temperature had a profound effect on the postthaw developmental competence of MII human oocytes in vitro and in vivo. The CryoLoop vitrification of first polar body-biopsied MII human oocytes in the presence of 5 mol/L ethylene glycol plus 1.3 mol/L dimethyl sulfoxide gave the best results in terms of fertilization, embryo development, and implantation rates.


FACS-Assisted CRISPR-Cas9 Genome Editing Facilitates Parkinson's Disease Modeling.
Genome editing and human induced pluripotent stem cells hold great promise for the development of isogenic disease models and the correction of disease-associated mutations for isogenic tissue therapy. CRISPR-Cas9 has emerged as a versatile and simple tool for engineering human cells for such purposes. However, the current protocols to derive genome-edited lines require the screening of a great number of clones to obtain one free of random integration or on-locus non-homologous end joining (NHEJ)-containing alleles. Here, we describe an efficient method to derive biallelic genome-edited populations by the use of fluorescent markers. We call this technique FACS-assisted CRISPR-Cas9 editing (FACE). FACE allows the derivation of correctly edited polyclones carrying a positive selection fluorescent module and the exclusion of non-edited, random integrations and on-target allele NHEJ-containing cells. We derived a set of isogenic lines containing Parkinson's-disease-associated mutations in α-synuclein and present their comparative phenotypes.


Measurement of both left ventricular function and regional myocardial perfusion with 133Xe in dogs.
A technique to measure left ventricular (LV) function and myocardial perfusion was validated in 12 dogs. 133Xe in saline was injected into the left atrium (LA) or LV and two data sets were obtained using gamma camera imaging: 1) A first pass gated scan for LV function; followed by 2) Sequential images for regional myocardial perfusion. LV ejection fraction and wall motion measurements from the 133Xe blood pool images were compared to ejection fraction (r = 0.88, P less than 0.01) and wall motion (r = 0.83, P less than 0.01) data from 99mTc labeled blood pool scans. The perfusion measurements obtained with the 133Xe method were compared to microsphere data (r = 0.79, P less than 0.01). Measurements after LV 133Xe injection were similar to data following LA injection. Thus, quantitative assessment of global LV function, regional wall motion and myocardial perfusion is possible with LA or LV 133Xe injection and gamma camera imaging.


Dyspnea and dysphagia associated to hypopharyngeal fibrolipoma: A case report.
Fibrolipomas are benign lesions conformed by fat and connective tissue, classified as histologic variants of lipomas. They are rarely located in the head and neck and represent less than 0.6% of the benign tumors of the larynx and hypopharynx. Their clinical presentation depends on its location and size. We present the case of a 51-year-old male patient who reported progressive dyspnea, dysphagia and obstructive sleep symptoms with a duration of 3 months, without apparent cause. A pharyngolaryngeal fiberoptic endoscopy showed a smooth, rounded mass in the posterior wall of the hypopharynx, partially obstructing the laryngeal vestibule, creating a valve effect. Complete trans-cervical resection of the lesion was performed after the airway was secured by means of a tracheotomy. The final histopathology report was fibrolipoma. He is currently asymptomatic and without evidence of relapse one year after the procedure.


Stimulating factors for regulation of osteogenic and chondrogenic differentiation of mesenchymal stem cells.
Mesenchymal stem cells (MSCs), distributed in many tissues in the human body, are multipotent cells capable of differentiating in specific directions. It is usually considered that the differentiation process of MSCs depends on specialized external stimulating factors, including cell signaling pathways, cytokines, and other physical stimuli. Recent findings have revealed other underrated roles in the differentiation process of MSCs, such as material morphology and exosomes. Although relevant achievements have substantially advanced the applicability of MSCs, some of these regulatory mechanisms still need to be better understood. Moreover, limitations such as long-term survival in vivo hinder the clinical application of MSCs therapy. This review article summarizes current knowledge regarding the differentiation patterns of MSCs under specific stimulating factors.


Malignant sacrococcygeal germ cell tumors in childhood: The Associazione Italiana Ematologia Oncologia Pediatrica (AIEOP) experience.
To evaluate clinical features at diagnosis, prognostic factors, and outcomes of malignant sacrococcygeal germ cell tumors (SC-GCTs) in patients enrolled in the Associazione Italiana Ematologia Oncologia Pediatrica (AIEOP) TCG 2004 protocol. A prospective analysis was conducted on all consecutive patients diagnosed with malignant SC-GCTs between January 2004 and May 2017. Patients with stage I underwent surgery and subsequent surveillance, the others received pediatric cisplatinum-etoposide-bleomycin (pPEB) regimen and eventual deferred surgery. Of 45 patients, 35 were females. Age at diagnosis ranged from 1 day to 3.6 years (median 1.6 years); 26 were stage IV. Of 38 patients who underwent surgery, pathology revealed yolk sac tumor (YST) in 27 and teratoma + YST/embryonal carcinoma in 11, while seven patients were diagnosed based on imaging and elevated levels of alpha-fetoprotein (AFP). Of six patients approached with surgery, only one relapsed and was rescued with first-line chemotherapy. Overall, 38 out of 45 achieved complete remission, three a partial remission, and four were resistant. Ten out of 41 patients who entered remission later relapsed and nine were rescued with a second-line treatment. We observed a global failure percentage of 31% and a 5-year overall survival (OS) and event-free survival (EFS) of 95% and 69%, respectively. Chemotherapyis generally effective in malignant SC-GCTs, even though almost one-third of our patients experienced events salvageable with second-line treatment. Most of the relapses occurred within 1 year from diagnosis. A close follow up with serial AFP level monitoring should be done for at least 2 years after diagnosis.


The effect of hexamethonium on gastric acid secretion in the conscious rat.
Evidence against a role for histamine in pentagastrin-stimulated acid secretion has been obtained previously using hexamethonium in gastric fistula rats. This possibility has been re-examined in conscious rats provided with gastric fistulae or Heidenhain pouches. Hexamethonium (20 mg/kg s.c.) inhibited basal acid secretion and acid secretion stimulated by histamine and pentagastrin in gastric fistula rats. The same dose of hexamethonium failed to produce a significant inhibition of acid secretion stimulated by bethanechol, pentagastrin or histamine in the presence of a low dose of bethanechol in Heidenhain pouch rats. These results provide no evidence to oppose the view that pentagastrin-stimulated acid secretion in the rat is mediated at least in part through the mobilization of gastric mucosal histamine. The inhibition of secretagogue induced acid secretion in the gastric fistula rat is mainly the result of a reduction in the basal acid output.


Functional efficiency of MerA protein among diverse mercury resistant bacteria for efficient use in bioremediation of inorganic mercury.
MerA protein of mer operon in mercury resistant bacteria influences transformation of Hg2+ to Hg0. Both in-silico and in-vivo studies have been carried out and MerA sequences, conserved motifs for mercury binding and NADPH (GCVPSK and LSCCA) varied widely in both Gram-positive and Gram-negative bacteria. As MerA-NADPH-FAD complex plays an important role in mercury volatilization, molecular interaction studies between MerA, NADPH, FAD and Hg2+ was carried out to study the efficiency of transformation of Hg2+ to Hg0 in mercury resistant bacteria. After the prediction of suitable models and molecular interaction analysis, the potential energies in the selected bacteria were as follows: Bacillus thuringiensis (NADPH: -5.15 kcal/mol and FAD: -9.63 kcal/mol), Pseudomonas aeruginosa (NADPH: -3.8 kcal/mol and FAD: -8.56 kcal/mol), Exiguobacterium sp. (NADPH: -3.37 kcal/mol and FAD: -8.42 kcal/mol), Vibrio sp. (NADPH: -3.3 kcal/mol and FAD: -6.7 kcal/mol) and Escherichia coli (NADPH: -3.28 kcal/mol and FAD: -5.69 kcal/mol). Additionally, the binding scores between MerA and Hg2+ followed the similar trend and found higher in B. thuringiensis (3.79) followed by P. aeruginosa (3.57), Exiguobacterium sp. (2.37), Vibrio sp. (1.47) and E. coli (1.07). ANOVA (2-way) result showed the significant (P < 0.05) variation among the energy values obtained after interaction studies. In-vivo analysis of expression of merA gene and Hg2+ removal efficiency also followed the same pattern with a highly significant correlation (P < 0.001) between the binding energy, binding score and expression pattern of merA gene as well as Hg2+ volatilization. Thus, the mercury removal efficiency of bacteria is genera specific which is correlated with the binding efficiency between MerA-NADPH complex and Hg2+ in mer operon mediated mercury resistant bacteria.


Choline acetyltransferase: regulation and coupling with protein kinase and vesicular acetylcholine transporter on synaptic vesicles.
Both the membrane-bound choline acetyltransferase (MChAT) and soluble ChAT (SChAT) were found to be activated by ATP-mediated protein phosphorylation. ATP activation of MChAT but not SChAT was found to depend on the integrity of proton gradient of synaptic vesicles because conditions disrupting the proton gradient also abolished the activation of MChAT by ATP. Among the kinases studied, Ca2+/calmodulin kinase II is most effective in activation of MChAT. Transport of ACh into synaptic vesicles by vesicular acetylcholine transporter (VAChT) is also proton gradient-dependent; therefore we proposed that there is a functional coupling between ACh synthesis and its packaging into synaptic vesicles. This notion is supported by the following findings: first, the newly synthesized [3H]-ACh from [3H]-choline was taken up much more efficiently than the pre-existing ACh; second, ATP-activation of MChAT was abolished when VAChT was inhibited by the specific inhibitor vesamicol; third, the activity of ChAT was found to be markedly increased when neurons are under depolarizing conditions.


Elevated plasma succinate in PTEN, SDHB, and SDHD mutation-positive individuals.
Cowden syndrome results from germline mutations in the gene for phosphatase and tensin homologue deleted on chromosome 10 (PTEN) and from variants in succinate dehydrogenase B and D subunits. We hypothesized that succinate accumulation may be common among individuals with SDH variants/mutations and those with PTEN mutations. Urine and blood were collected from individuals meeting full or partial Cowden syndrome diagnostic criteria or those with paraganglioma (PGL) or a known susceptibility paraganglioma-associated gene mutation, and succinate was measured. PTEN, SDHB, SDHC, and SDHD genes were sequenced from genomic DNA. Elevated plasma succinate was observed in 13/21 (62%) individuals with germline PTEN, SDHB, or SDHD mutations as compared with 5/32 (16%) controls (P < 0.001), in 10/15 (67%) individuals with pathogenic PTEN mutations but in <20% of mutation-negative individuals meeting identical criteria, and in individuals with mutations in SDHB (1/1, 100%) and SDHD (2/5, 40%). Our data suggest that mutations in PTEN, SDHB, and SDHD reduce catalytic activity of succinate dehydrogenase, resulting in succinate accumulation, and identify a common biochemical alteration in these two patient populations (PTEN and SDHx mutation positive individuals). Plasma organic acid analysis may provide an effective and inexpensive screening method to determine when more expensive gene sequencing of PTEN and SDH genes is warranted.


Morphometrical and Immunohistochemical Evaluation of Kidney as an Indirect Parameter to Estimate Age in Puppies in Veterinary Forensic Pathology.
Estimation of age represents a central focus in the veterinary forensic pathology field. Currently, the visual examination of the dentition and the skeletal age are the main methods to estimate the age of puppies. Nevertheless, these methods are affected by a broad range of variables. In contrast, the kidney is characterized by a specific postnatal development. In human glomerulogenesis, fetal mesangial cells change their immunohistochemical phenotypes with maturation. Therefore, we hypothesized that histological and immunohistochemical examinations of the kidney can be used together as an indirect parameter for age determination in puppies' cadavers. Forty-five puppies' cadavers were divided into five groups defined by age (Group A= 0-15 days, Group B = 16-45 days, Group C = 46-85 days, Group D = 86-105 days, Group E= 105-365 days). For each case, kidney samples were collected and processed for histopathological (for morphometrical study of the glomerulus) and immunohistochemical (for the immunolocalization of the α-SMA protein) studies. Morphometrical study allowed us to observe statistical differences in the mean glomerulus numbers per field among assessed groups. Similarly, immunohistochemical examination showed differences in SMA expression among groups. Our findings suggest a potential use of kidney morphometrical and immunohistochemical examinations together as an indirect parameter to assess the age of illegally imported puppies.


Bitterness and toxicity in wild yam (Dioscorea spp.) tubers of Nepal.
Wild yams make a significant contribution to diets of tribal people in Nepal. However, these wild tubers are unpalatable, taste bitter, produce inflammation and show occasional toxicity. Four wild yam species, which are eaten after primary treatment by Nepali aborigines, were analyzed for bitter and toxic principles. Bitter components were identified as furanoid norditerpenes (diosbulbins A and B). Diosbulbins A and B were found in the range of 0.023-0.046 and 0.151-0.442 g kg(-1), respectively. Results demonstrated that diosbulbin B, with an average value of 0.314 g kg(-1), was the principal bitter compound as compared to diosbulbin A (0.037 g kg(-1)). The toxic alkaloid, dioscorine and histamine (an allergen) were not detected in these tubers, whereas cyanogens (as HCN equivalent) content were found ranging from 3.2 to 6.0 ppm. Our results revealed that Nepali wild yam tubers are not toxic varieties, as they do not contain either toxic dioscorine or histamine and cyanogens contents were satisfactorily below the safety limits. The inflammation and occasional toxicity observed could possibly be due to the presence of high level of oxalate in these tubers. Domestic cooking methods were found to be very efficient in removing bitterness, thus making the bitter yams palatable.


The COOP/WONCA charts in an acute psychiatric ward. Validity and reliability of patients' self-report of functioning.
We wanted to examine whether persons needing acute psychiatric admittance can give us reliable information about their functional status with the COOP/WONCA charts (Dartmouth Primary Care Cooperative Information Project-World Organization of National Colleges, Academies and Academic Associations of General Practitioners/Family Physicians), and if this information parallel that of therapists and ward staff. To examine internal consistency, external reliability and test-retest reliability, all patients consecutively admitted to an acute psychiatric department were asked to fill in the COOP/WONCA, Short-Form-36 Health Survey (SF-36) and Symptom Checklist 90--Revised (SCL-90-R) 1-2 weeks after admittance. Their therapists scored a special version of the GAF, and the ward staff scored an observer version of the COOP/WONCA. The first 59 patients repeated the COOP/WONCA scoring after a few days to estimate of test-retest reliability. Of a total of 267 persons admitted in the study period, 102 were included. Non-inclusion was largely due to early discharge. The internal consistency of the COOP/WONCA corresponded to a Cronbach's alpha=0.85. External reliability versus the SF-36 resulted in a correlation of r=-0.82 (P<0.001), and versus the observer version of COOP/WONCA we found r=0.46 (P<0.001). Correlation with GAF was low and not significant. The COOP/WONCA correlated significantly with the SCL-90-R scores (r=0.59, P<0.001). The test-retest correlation was r=0.85 (P<0.001). Correlations varied between sub-groups of patients, but all had consistently high correlations between the various self-scored measures. The COOP/WONCA gives a consistent and reliable report of acutely admitted psychiatric inpatients' own views of their functional capacity. The therapists' views and to some degree the ward staff's views diverge from the patients' opinions, especially for the more seriously disturbed patients.


Tiaprofenic acid. A review of its pharmacological properties and therapeutic efficacy in rheumatic diseases and pain states.
Tiaprofenic acid is a new non-steroidal anti-inflammatory agent advocated for use in rheumatoid arthritis, osteoarthritis, musculoskeletal disorders, soft-tissue injuries and inflammatory conditions and acute pain of varying origin. Published data suggest that tiaprofenic acid 600 mg daily in 2 or 3 divided doses is comparable in effectiveness with aspirin, diclofenac, ibuprofen, indomethacin, naproxen, piroxicam and sulindac in the treatment of rheumatoid arthritis and osteoarthritis. More controlled clinical trials are necessary to evaluate its potential in rheumatic conditions other than rheumatoid arthritis and osteoarthritis. In controlled studies in patients with acute pain following surgery or trauma, tiaprofenic acid was more effective than placebo and as effective as aspirin and indomethacin. While tiaprofenic acid produced fewer side effects than aspirin in rheumatoid arthritis treatment, and indomethacin in the treatment of osteoarthritis, results have generally shown the short term tolerability of tiaprofenic acid to be similar to that of other non-steroidal anti-inflammatory drugs. As no one of the non-steroidal anti-inflammatory agents is the most suitable drug for all patients requiring such therapy, tiaprofenic acid should be considered along with other drugs of this type in the therapy of arthritic conditions and of acute postoperative or posttraumatic pain.


Hemodynamic alterations in the transferred tissue to lower extremities.
A higher incidence of failure has been reported for free flaps transplanted to the lower extremities. However, the physiological background of this phenomenon has not been elucidated. We reviewed the 3-day postoperative hemodynamic data for 103 free flaps, including the in situ venous pressure (N = 103), arterial pressure (N = 53), and surface blood flow (N = 42). The cases were divided into two groups based on the recipient site, i.e., lower extremity (the LE group: N = 29) and the other (non-LE group: N = 74). The venous pressure was significantly higher in the LE group (26.6 +/- 2.2 vs. 14.8 +/- 1.2 mmHg), whereas the arterial pressure immediately after surgery was lower than the non-LE group. The hemodynamic data within the transferred tissues demonstrated significant differences between groups, especially in the early postoperative period. There is a possibility that the high venous pressure may aggravate the poor perfusion in tissues transferred to the lower extremities.


Factors defining the mentoring competencies of clinical midwives: An exploratory quantitative research study in Japan.
Clinical education is an extremely important process in cultivating healthcare professionals, and the quality of educators has a major impact on the quality of future practitioners. Although practicing clinical midwives contribute to the education of pre-registered midwives and those qualified within the past year (new midwives), the factors defining the educational competencies of clinical midwives have not been clarified. The purpose of this study was to explore the factors that define the mentoring competencies of clinical midwives involved in educating new midwives. An exploratory quantitative research study. Questionnaires were distributed to 694 midwives who had previously conducted educational activities with new midwives at the 63 facilities whose administrator or nurse manager in charge of all staff, including midwives, consented to participate. Of the 694 midwives, 464 (66.9%) returned the questionnaire and 451 (65.1%) valid responses were analyzed. Exploratory factor analyses were performed on the following three concepts: [competency as a professional], [competency as an educator], and [personal characteristics]. [Competency as a professional] consisted of two factors: <self-awareness and self-reflection for finding confidence> and <sharing their midwifery practice>; [competency as an educator] consisted of four factors: <supporting experimental study>, <thoughtfulness and empathy for new midwives>, <making effective use of new midwives' own experience> and <commitment to educational activities>; and [personal characteristics consisted of three factors: <exercising leadership> <approachability>, and <tendency to be a caring mentor>. These three concepts were defined by a total of nine sub-concepts (factors), and 41 items were extracted with a reliability coefficient (Cronbach's α) of 0.944 CONCLUSIONS: "Mentoring competencies of clinical midwives (MCCM)" are defined by three concepts and nine sub-concepts, which can be evaluated by 41 items regarding the behavior, thoughts, and characteristics that clinical midwives exhibit when they educate new midwives in clinical settings.


Bacteria- and temperature-regulated peptides modulate β-catenin signaling in Hydra.
Animal development has traditionally been viewed as an autonomous process directed by the host genome. But, in many animals, biotic and abiotic cues, like temperature and bacterial colonizers, provide signals for multiple developmental steps. Hydra offers unique features to encode these complex interactions of developmental processes with biotic and abiotic factors, and we used it here to investigate the impact of bacterial colonizers and temperature on the pattern formation process. In Hydra, formation of the head organizer involves the canonical Wnt pathway. Treatment with alsterpaullone (ALP) results in acquiring characteristics of the head organizer in the body column. Intriguingly, germfree Hydra polyps are significantly more sensitive to ALP compared to control polyps. In addition to microbes, β-catenin-dependent pattern formation is also affected by temperature. Gene expression analyses led to the identification of two small secreted peptides, named Eco1 and Eco2, being up-regulated in the response to both Curvibacter sp., the main bacterial colonizer of Hydra, and low temperatures. Loss-of-function experiments revealed that Eco peptides are involved in the regulation of pattern formation and have an antagonistic function to Wnt signaling in Hydra.


First report of Blastocystis infections in cattle in China.
Blastocystis is one of the most common intestinal protists of humans and can also infect a variety of other mammals and birds. Blastocystis infections and subtype distribution in cattle have been documented, while nothing is known about those in China. Herein, a total of 526 dairy cattle from northeast China were sampled and investigated for the prevalence and genetic characteristics of Blastocystis and the potential role of bovine animals in zoonotic transmission of Blastocystis. The parasite was identified in 54 (10.3%) fecal specimens by nested PCR and DNA sequencing of the small subunit ribosomal RNA gene. Sequence analysis enabled identification of four Blastocystis subtypes (STs). Among those, subtype ST10 (75.9%, 41/54) has the highest frequency, followed by ST14 (18.5%, 10/54), ST4 (3.7%, 2/54), and ST5 (1.9%, 1/54). High prevalence and widespread distribution of ST10 and ST14 in cattle observed herein, together with analysis of their host distribution patterns in earlier studies, indicated some host-adapted potential in the two subtypes. The identification of human-pathogenic subtypes ST4 and ST5 might imply a potential zoonotic risk of cattle origin. This is the first study exploring the prevalence and genetic characteristics of Blastocystis in cattle in China. The host range of subtype ST4 was extended. The findings of this study should be helpful for a better understanding of the epidemiology and public health potential of Blastocystis.


Synthesis, conformational and spectroscopic characterization of monomeric styrene derivatives having pendant p-substituted benzylic ether groups.
Three derivatives of styrene monomer, 4-chlorophenyl-4-vinylbenzyl ether (I), 4-methoxyphenyl-4-vinylbenzyl ether (II) and 4-ethylphenyl-4-vinylbenzyl ether (III) were synthesized. The synthesized two novel compounds (I and III) and one with undefined structural features were identified by experimental spectroscopic techniques and density functional approach. The optimized geometrical structure, vibrational and electronic transitions along with chemical shifts of those compounds were presented in this study. The vibrational spectra of investigated compounds were recorded in solid state with FT-IR spectrometry in the range of 4000-400 cm(-1). The computational vibrational wavenumbers and also ground state equilibrium conformations were carried out by using density functional method with 6-311++G(d,p) basis set. Assignments of the fundamental vibrational modes were examined on the basis of the measured data and total energy distribution (TED) of the vibrational modes, calculated with scaled quantum mechanical (SQM) method. Isotropic chemical shift of hydrogen and carbon nuclei were investigated via observed (1)H and (13)C NMR spectra in deuterated DMSO solution and predicted data applied with gauge-invariant atomic orbitals (GIAOs) method. The UV absorption spectra of monomers were observed in the range of 200-800 nm in ethanol, and time dependent DFT method was used to obtain the electronic properties. A detailed description of spectroscopic behaviors of compound was given based on the comparison of experimental measurements and theoretical computations.


Synthesis of iodinated 3beta-aryltropanes with selective binding to either the dopamine or serotonin transporters.
Iodinated 3beta-aryltropanes functionalized appropriately at the 2beta-, 8- and aryl positions display selective binding to either the dopamine or serotonin transporters.


Adenine methylation in zein genes.
This paper reports the novel finding of adenine methylation in higher plants. Comparison of restriction patterns of genomic maize DNA digested with enzymes MboI and Sau3A enabled us to detect the existence of adenine methylation in zein genes. Adenine methylation within or around zein genes turned out to be similar in endosperm (where zeins are actively synthesized) and in seedling tissue (where zein genes are not expressed). Furthermore, adenine methylation patterns were found to be similar both in wild-type and opaque-2 mutant plants. These lines of evidence suggest that adenine methylation is unrelated to the regulation of gene expression.


Genetic variant RNF213 c.14576G>A in various phenotypes of intracranial major artery stenosis/occlusion.
Recently, we reported a common genetic variant, ring finger protein 213 (RNF213) c.14576G>A variant, a susceptibility gene for moyamoya disease (MMD), among patients with intracranial major artery stenosis/occlusion (ICASO) in a selected Japanese population. The aim of this 2-center-based case-control study was to confirm our previous finding in a larger population. Study participants were recruited from The University of Tokyo Hospital and Kanto Neurosurgical Hospital. The occurrence rate of c.14576G>A variant was investigated in 323 patients, 22 with definite MMD, 8 with unilateral MMD, 84 with ICASO in the absence of MMD (non-MMD ICASO), 34 with extracranial carotid atherosclerosis, 44 with cerebral aneurysm, 21 with intracerebral hemorrhage, and 110 control subjects. RNF213 c.14576G>A variant was found in 1.8% (2/110) of the normal control group and had significant associations with definite MMD (P<0.0001; odds ratio, 144.0; 95% confidence interval, 26.7-775.9), unilateral MMD (P=0.0001; odds ratio, 54.0; 95% confidence interval, 7.5-386.8), and non-MMD ICASO (P<0.0001; odds ratio, 16.8; 95% confidence interval, 3.81-74.5). There was no significant association with extracranial carotid atherosclerosis, cerebral aneurysm, or intracerebral hemorrhage. This result replicated our previous findings. A particular subset of patients with various phenotypes of ICASO has a common genetic variant, RNF213 c.14576G>A, indicating that RNF213 c.14576G>A variant is a high-risk allele for ICASO.


Challenging declarations of abstinence by the determination of morphine in hair by radioimmunoassay.
Morphine extracted from dated hair segments and analysed by adaptation of a specific solid-phase 125I radioimmunoassay for the measurement of morphine in urine has provided long term histories of the heroin usage of individual patients. Results expressed as ng morphine/mg hair are compared with a pre-determined cut-off level of 0.3 ng morphine/mg hair, from a known drug-free population (n = 21). Morphine concentrations in hair samples from known heroin abusers are found to be above the cut-off limit (range 0.6-9.4 ng morphine/mg hair), and suggest a possible correlation between heroin intake and concentration of morphine in hair. Intra-assay relative standard deviation (RSD), at morphine levels of 9.27 and 1.12 ng morphine/mg hair are 2.4 and 5.5%, respectively, and acceptable recoveries from drug-free hair spiked with morphine are also achieved. The developed segmental hair analysis regime has been used successfully to challenge a self-declaration of heroin abstinence. Routine cumulative urine screening was unable to provide such evidence. The potential for hair drug analysis is discussed.


Proliferation of microbodies in Saccharomyces cerevisiae.
The development of microbodies in the yeast Saccharomyces cerevisiae was studied in response to different conditions of growth. Various strains of S. cerevisiae were investigated, using cells from the exponential growth phase on glucose as an inoculum in all transfer experiments. Electron microscopy, including serial sectioning, revealed that these cells generally contained one to four small microbodies which were localized in the vicinity of the cell wall and characterized by the presence of catalase. Transfer of these glucose-grown cells into media supplemented with various compounds known to induce microbody proliferation in other yeasts--i.e. uric acid, alkylated amines, amino acids, C2-compounds such as ethanol or acetate, in the presence or absence of compounds that induce oxygen radical formation--did not result in a significant change in the number of microbody profiles observed. Marked microbody proliferation was, however, observed after a shift of cells into media containing oleic acid and was associated with the induction of activities of beta-oxidation enzymes. In addition, catalase and isocitrate lyase were present in enhanced levels. Kinetic experiments suggested that these microbodies developed from those originally present in the inoculum cells. In thin sections up to 14 microbody profiles were occasionally observed, often present in small clusters. Their ultimate volume fraction amounted to 8-10% of the cytoplasmic volume.


A new method based on additive vegetation index for mapping Huangtai algae coverage in Lake Ulansuhai.
Huangtai algal blooms are key indicators of eutrophication and lake-ecosystem damage. Understanding the spatiotemporal heterogeneity of their growth is critical for preserving the ecological environment. The dimidiate pixel model is commonly used to estimate vegetation coverage; however, indices such as the normalized difference vegetation index have not been specifically constructed for the Huangtai algae spectrum and thus are not specific or sufficiently precise for use as indicators. Therefore, we propose a new dimidiate pixel model based on a novel additive vegetation index to calculate the Huangtai algal coverage for each pixel using Landsat multispectral satellite images with 30-m resolution. The results showed that the additive vegetation index with R2 = 0.994 is a better indicator than the normalized difference vegetation index, enhanced vegetative index, and ratio vegetative index, with the accuracy of the new model reaching 86.61%. Monthly Landsat images from 2006 to 2016 were used to calculate the Huangtai algal coverage. Analysis of the inter-monthly variation indicated increased coverage from May to July, with an annual maximum and minimum of 14.43% and 0.33% in 2008 and 2013, respectively. This study provides a new reference map of Huangtai algal cover, which is important for monitoring and protecting the Lake Ulansuhai environment.


Reactivity of islet cell antibodies (ICA) to recombinant glutamic acid decarboxylase (GAD).
To study the heterogeneity of islet cell antibodies (ICA), recombinant rat and human GAD65 expressed as bacterial fusion proteins were used to inhibit ICA reactivity in sera from recent onset type 1 diabetic children and ICA-positive first degree relatives of diabetic patients. Rat GAD65 was expressed as a fusion protein in the expression vector RSET and inhibited ICA (GAD+ICA) in 26% of 23 recent onset patients, 29% of 14 ICA positive first degree relatives (FDR) who progressed to diabetes (prediabetics) and 50% of 20 FDR who did not progress to diabetes 18 months to 5 years (31 +/- 14 months) after collection of the sample. GAD+ICA were inversely associated with the presence of insulin autoantibodies (IAA) (P = 0.006). GAD antibodies (GAD-Ab) were also detected by immunoprecipitation of in vitro transcribed and translated [35S] methionine-labelled human GAD65. GAD-Ab were present in 83% of recent onset patients, 86% of prediabetics and 95% of the relatives who did not progress to diabetes. The level of GAD-Ab was higher in the presence of GAD+ICA (1.39 +/- 0.57 vs 0.79 +/- 0.6 index units; P = 0.001). ICA levels were higher in GAD-Ab negative than in GAD-Ab positive sera (377 +/- 256 vs 195 +/- 231 JDFU; P = 0.03). Our results confirm that a recombinant GAD65 fusion protein can be used to detect ICA heterogeneity. However, neither inhibition of ICA with recombinant GAD, nor direct detection of GAD-Ab improved the prediction of progression to clinical diabetes in ICA positive FDR.


[Effect of L-DOPA on brain serotonin metabolism related to tryptophan concentrations in plasma and brain of rats].
Changes in tryptophan (Trp) concentrations in the peripheral and brain tissues and their influences on brain serotonin (5-HT) metabolism were investigated after i.p. administrations of L-DOPA alone and in combination with Ro4-4602 in male Wistar rats. L-DOPA alone elicited marked increases of both plasma free and liver Trp levels, and a slight increase of brain Trp level. Concomitant application of L-DOPA and Ro4-4602 did not change peripheral Trp concentrations but did reduce brain Trp level. Brain 5-HT levels were reduced in proportion to the increase of dopamine (DA) level by L-DOPA alone and Ro4-4602 plus L-DOPA. A marked increase of 5-HIAA level was exhibited by L-DOPA alone, while Ro4-4602 plus L-DOPA increased 5-HIAA levels slightly despite a marked decrease in 5-HT levels. The decrease of 5-HT and increase of 5-HIAA levels were also observed after administration of L-DOPA to neonatal rats in which the brain tryptophan hydroxylase was considered to be saturated. These results suggest that changes in 5-HT metabolism after L-DOPA injections are not only caused by DA which displaces 5-HT from vesicular stores, but also by changes in Trp concentrations in the plasma and brain which alter 5-HT synthesis and turnover.


Evaluating the use of a balance prosthesis during balance perturbations in children and young adults with cochleovestibular dysfunction.
Study objectives were to: (1) quantify stability in children and young adults using cochlear implants with concurrent cochleovestibular dysfunction (CI-V) during balance perturbations and (2) to assess effects of an auditory head-referencing device (BalanCI) on their stability. The BalanCI provides auditory feedback via cochlear implants to cue posture and potentially avoid falling in children with CI-V. It was hypothesized that children and young adults with CI-V respond with larger movements to floor perturbations than typically-developing peers (controls) and that BalanCI use decreases these movements. Motion in response to treadmill perturbations was captured by markers on the head, torso, and feet in eight CI-V and 15 control participants. Stability (area under the curve of motion displacement) and peak displacement latencies were measured. The CI-V group demonstrated less stability and slower responses than the control group during medium and large backwards perturbations (p's < 0.01). In the CI-V group, BalanCI use improved stability during large backwards perturbations (p < 0.001), but worsened stability during large sideways perturbations (p's < 0.001). Children and young adults with CI-V move more to remain upright during perturbations than typically-developing peers. The BalanCI has potential to aid physical/vestibular therapy in children with CIs who have poor balance.


The asymmetric segregation of parental nucleosomes during chrosome replication.
SV40 DNA replicated in the presence of cycloheximide was more sensitive to staphylococcal nuclease digestion and had a lower superhelical density than viral DNA replicated in the absence of this drug. These data indicate that fewer nucleosomes are associated with progeny SV40 DNA molecules after DNA replication in the absence of protein synthesis and that these nucleosomes are derived from the parental histones. We designed an experiment to determine whether these parental SV40 nucleosomes segregate to the leading side of the replication form where DNA synthesis is continuous, the lagging side of the fork where DNA synthesis is discontinuous or randomly to both sides of the fork. The results indicate that the parental histones distributed themselves asymmetrically, preferentially (80-90%) segregating with the leading side of both SV40 DNA replication forks during bidirectional replication in the absence of protein synthesis. In the case of SV40, the same parental DNA strands are the templates for the leading side of DNA replication at both forks as well as the templates for the informational or coding strand of early and late viral mRNA synthesis. Based on this correspondence, we designed an experiment to test whether chicken cells growing in culture and replicating their DNA in the absence of protein synthesis segregated their parental histones asymmetrically to the progeny DNA strand that also coded for stable nuclear RNA transcripts. The results of these experiments indicate that, like SV40, parental cellular histones segregate asymmetrically and are preferentially associated with those DNA template strands that code for stable nuclear RNA species detected by hybridization to single-copy DNA.


Automated 3D U-net based segmentation of neonatal cerebral ventricles from 3D ultrasound images.
Intraventricular hemorrhaging (IVH) within cerebral lateral ventricles affects 20-30% of very low birth weight infants (<1500 g). As the ventricles increase in size, the intracranial pressure increases, leading to post-hemorrhagic ventricle dilatation (PHVD), an abnormal enlargement of the head. The most widely used imaging tool for measuring IVH and PHVD is cranial two-dimensional (2D) ultrasound (US). Estimating volumetric changes over time with 2D US is unreliable due to high user variability when locating the same anatomical location at different scanning sessions. Compared to 2D US, three-dimensional (3D) US is more sensitive to volumetric changes in the ventricles and does not suffer from variability in slice acquisition. However, 3D US images require segmentation of the ventricular surface, which is tedious and time-consuming when done manually. A fast, automated ventricle segmentation method for 3D US would provide quantitative information in a timely manner when monitoring IVH and PHVD in pre-term neonates. To this end, we developed a fast and fully automated segmentation method to segment neonatal cerebral lateral ventricles from 3D US images using deep learning. Our method consists of a 3D U-Net ensemble model composed of three U-Net variants, each highlighting various aspects of the segmentation task such as the shape and boundary of the ventricles. The ensemble is made of a U-Net++, attention U-Net, and U-Net with a deep learning-based shape prior combined using a mean voting strategy. We used a dataset consisting of 190 3D US images, which was separated into two subsets, one set of 87 images contained both ventricles, and one set of 103 images contained only one ventricle (caused by limited field-of-view during acquisition). We conducted fivefold cross-validation to evaluate the performance of the models on a larger amount of test data; 165 test images of which 75 have two ventricles (two-ventricle images) and 90 have one ventricle (one-ventricle images). We compared these results to each stand-alone model and to previous works including, 2D multiplane U-Net and 2D SegNet models. Using fivefold cross-validation, the ensemble method reported a Dice similarity coefficient (DSC) of 0.720 ± 0.074, absolute volumetric difference (VD) of 3.7 ± 4.1 cm3 , and a mean absolute surface distance (MAD) of 1.14 ± 0.41 mm on 75 two-ventricle test images. Using 90 test images with a single ventricle, the model after cross-validation reported DSC, VD, and MAD values of 0.806 ± 0.111, 3.5 ± 2.9 cm3 , and 1.37 ± 1.70 mm, respectively. Compared to alternatives, the proposed ensemble yielded a higher accuracy in segmentation on both test data sets. Our method required approximately 5 s to segment one image and was substantially faster than the state-of-the-art conventional methods. Compared to the state-of-the-art non-deep learning methods, our method based on deep learning was more efficient in segmenting neonatal cerebral lateral ventricles from 3D US images with comparable or better DSC, VD, and MAD performance. Our dataset was the largest to date (190 images) for this segmentation problem and the first to segment images that show only one lateral cerebral ventricle.


The presence of CYP79 homologues in glucosinolate-producing plants shows evolutionary conservation of the enzymes in the conversion of amino acid to aldoxime in the biosynthesis of cyanogenic glucosides and glucosinolates.
A cDNA encoding CYP79B1 has been isolated from Sinapis alba. CYP79B1 from S. alba shows 54% sequence identity and 73% similarity to sorghum CYP79A1 and 95% sequence identity to the Arabidopsis T42902, assigned CYP79B2. The high identity and similarity to sorghum CYP79A1, which catalyses the conversion of tyrosine to p-hydroxyphenylacetaldoxime in the biosynthesis of the cyanogenic glucoside dhurrin, suggests that CYP79B1 similarly catalyses the conversion of amino acid(s) to aldoxime(s) in the biosynthesis of glucosinolates. Within the highly conserved 'PERF' and the heme-binding region of A-type cytochromes, the CYP79 family has unique substitutions that define the family-specific consensus sequences of FXP(E/D)RH and SFSTG(K/R)RGC(A/I)A, respectively. Sequence analysis of PCR products generated with CYP79B subfamily-specific primers identified CYP79B homologues in Tropaeolum majus, Carica papaya, Arabidopsis, Brassica napus and S. alba. The five glucosinolate-producing plants identified a CYP79B amino acid consensus sequence KPERHLNECSEVTLTENDLRFISFSTGKRGC. The unique substitutions in the 'PERF' and the heme-binding domain and the high sequence identity and similarity of CYP79B1, CYP79B2 and CYP79A1, together with the isolation of CYP79B homologues in the distantly related Tropaeolaceae, Caricaceae and Brassicaceae within the Capparales order, show that the initial part of the biosynthetic pathway of glucosinolates and cyanogenic glucosides is catalysed by evolutionarily conserved cytochromes P450. This confirms that the appearance of glucosinolates in Capparales is based on a cyanogen 'predisposition'. Identification of CYP79 homologues in glucosinolate-producing plants provides an important tool for tissue-specific regulation of the level of glucosinolates to improve nutritional value and pest resistance.


Aciclovir and corneal wound healing.
Masked controlled rabbit studies were done to determine the toxic effects on corneal wound healing of the antiviral drugs 3% aciclovir and 0.5% idoxuridine ointment in therapeutically effective concentrations. Aciclovir had no significant detrimental effect in comparison to controls on the quality of regenerating epithelium or the re-epithelialization of epithelial wounds. Idoxuridine treatment caused significant toxic changes in the regenerating epithelium clinically and histologically with a significant delay in epithelial wound healing in comparison to control or aciclovir treated eyes. Aciclovir had no significant effect on the collagen content of stromal wounds as measured by hydroxyproline levels. Idoxuridine caused a reduction in collagen content not significantly different from controls but significantly lower than aciclovir.


Optimal assessment of lymph node status in gallbladder cancer.
Lymph node (LN) metastasis is an important prognostic factor in gallbladder cancer (GBCA). LN status has been adopted as a critical element of staging systems. However, the influence of total lymph node count (TLNC) remains unclear. We determined the optimal minimum TLNC and compared the prognostic significance of LN status indices in GBCA. We retrospectively reviewed medical records of 128 patients with T2 or greater GBCA who underwent LN dissection. We analyzed overall survival (OS) and relevance of the number of metastatic LNs, ratio of metastatic LNs to retrieved LNs (LNR), and TLNC in predicting OS. The median OS durations were 120, 35, and 18 months in T2, T3, and T4 GBCA. Five-year OS rates were 73%, 43%, and 0% in T2, T3, and T4 GBCA. LN status did not significantly impact OS in T2 or T4 GBCA. However, all LN indices were significantly correlated with OS in T3 GBCA. Furthermore, multivariate analysis revealed that a metastatic LN count of more than four and a TLNC of more than eight were independent prognostic factors of OS in T3 GBCA. TLNC and the number of positive LNs may be more important prognostic factors than LNR in T3 GBCA. Additionally, accurate staging may not be achieved in cases of T3 GBCA if the total number of retrieved LNs is less than eight. Thus, to ensure proper staging, we recommend that surgeons harvest more than eight LNs in patients with T3 GBCA.


Randomized controlled experiments in health and social sciences: some conceptual issues.
This brief article outlines some difficulties as well as benefits in conducting randomized controlled trials in social science settings especially in developing countries. Some of the historical developments are summarized and certain applications in health sciences are discussed from methodological and policy standpoints.


Schistosoma japonicum: an ultraviolet-attenuated cercarial vaccine applicable in the field for water buffaloes.
Water buffaloes were vaccinated three times with 10,000 Schistosoma japonicum cercariae irradiated with ultraviolet (uv) light at a dose of 400 microW x min/cm2. The irradiation was performed with cheap, simple, and portable equipment in a rural area of Hubei Province (People's Republic of China). A challenge infection of 1000 untreated cercariae was given to six vaccinated and six naive control buffaloes, while two vaccinated animals were not challenged. The experiment was terminated 6 weeks after the challenge. Control animals had lost body weight and harbored a mean of 110 worms and 37 eggs per gram of liver. The vaccinated animals gained weight after the challenge and developed 89% resistance to infection with S. japonicum. Since schistosomiasis japonica is nowadays transmitted in China predominantly by domestic livestock, a uv-attenuated cercarial vaccine for bovines may contribute to the control of this disease.


CARs: a new approach for the treatment of autoimmune diseases.
The development of chimeric antigen receptor (CAR)-based therapeutic interventions represented a breakthrough in cancer treatment. Following the success of the CAR-T-cell strategy, this novel therapeutic approach has been applied to other diseases, including autoimmune diseases. Using CAR-T cells to deplete pathological immune cells (i.e., B cells, autoreactive B or T cells, and accessory antigen-presenting cells (APCs)) has resulted in favorable outcomes in diseases characterized by excessive autoantibody levels or hyperactive lymphocyte cell numbers. The importance of immunosuppressive regulatory T cells (Tregs) in restoring immune tolerance has been well established, and CAR-Tregs have shown promising therapeutic potential in treating autoimmune diseases. Moreover, prior experience from the cancer field has provided sufficient paradigms for understanding how to optimize the structure and function of CARs to improve their function, persistence, stability and safety. In this review, we describe the potential application of CAR-T cells and CAR-Tregs in the treatment of autoimmune diseases, and we summarize the currently available strategies of gene editing and synthetic biological tools that have improved the practical application of CAR-based therapies.


Retrospective comparative study of the efficacy of JAK inhibitor (tofacitinib) in the treatment of systemic sclerosis-associated interstitial lung disease.
The oral Janus kinases inhibitor (JAKi) has improved the management of skin manifestations in systemic sclerosis (SSc), and our study aimed to explore the efficacy of non-selective JAKi tofacitinib in ameliorating interstitial lung disease (ILD) in the patients with SSc. The hospitalization data of the SSc-ILD patients from April 2019 to April 2021 were collected, and the changes of pulmonary function and the radiological findings in pulmonary high-resolution CT (HRCT) from the 9 patients who received tofacitinib for at least 6 months and a matched group of 35 SSc-ILD patients treated with conventional immunosuppressants or glucocorticoids, were compared and analyzed. There were no significant differences in demographic data and clinical characteristics between the tofacitinib-treated group (tofa-group) and the matched group. However, in the tofa-group, the changes in serum lactate dehydrogenase (LDH) concentration and serum interleukin-6 levels were significantly lower than those in the matched group. Moreover, the tofa-group showed amelioration in decreased diffusing capacity of the lung for carbon monoxide (DLCO) (62.05 ± 9.47 vs. 66.61 ± 12.39, p = 0.046), reductions in ground-glass attenuation involvement (1.00 ± 0.86 vs. 0.33 ± 0.50, p = 0.024) and irregular pleural thickening (1.33 ± 0.50 vs. 0.67 ± 0.51, p = 0.004) in pulmonary HRCTs, alleviated modified Rodnan skin score (mRSS) of skin sclerosis (9.22 ± 3.81 vs. 7.11 ± 3.92, p = 0.048), and reduced HRCT scores of pulmonary fibrosis (15.00 ± 3.87 vs. 12.66 ± 4.92, p = 0.009). Logistic regression analysis showed that the involvement of ground-glass attenuation (OR 11.43) and the add-on therapy of tofacitinib (OR 9.98) were the relevant factors in the amelioration of HRCT. Our results indicate that the use of JAKi (tofacitinib) may be relevant to significant improvement of the sclerosis and early radiological abnormalities in SSc-ILD patients. Further studies are needed to confirm these findings and to explore its efficacy more precisely. Key Points • The currently available therapies for SSc-ILD have limited therapeutic benefits. • The add-on therapy of the oral JAK inhibitor is available in the real world. • The tofacitinib was promising in the improvement of the sclerosis and early radiological abnormalities in SSc-ILD patients.


Reasons why patients fail screening in Indian breast cancer trials.
An increased number of screen failure patients in a clinical trial increases time and cost required for the recruitment. Assessment of reasons for screen failure can help reduce screen failure rates and improve recruitment. We collected retrospective data of human epidermal growth factor receptor (HER2) positive Indian breast cancer patients, who failed screening for phase 3 clinical trials and ascertained their reasons for screen failure from screening logs. Statistical comparison was done to ascertain if there are any differences between private and public sites. Of 727 patients screened at 14 sites, 408 (56.1%) failed screening. The data on the specific reasons for screen failures was not available at one of the public sites (38 screen failures out of 83 screened patients). Hence, after excluding that site, further analysis is based on 644 patients, of which 370 failed screening. Of these, 296 (80%) screen failure patients did not meet selection criteria. The majority -266 were HER2 negative. Among logistical issues, 39 patients had inadequate breast tissue sample. Sixteen patients withdrew their consent at private sites as compared to six at public sites. The difference between private and public sites for the above three reasons was statistically significant. Use of prescreening logs to reduce the number of patients not meeting selection criteria and protocol logistics, and patient counseling to reduce consent withdrawals could be used to reduce screen failure rate.


Present day controversies and consensus in curettage for giant cell tumor of bone.
The ideal treatment for giant cell tumor of bone (GCTB) is still controversial. The purpose of this study was to evaluate whether curettage was successful in the treatment of GCTB. Intralesional curettage with adjuvant therapies, such as high-speed burring, polymethylmethacrylate, phenol, ethanol, and liquid nitrogen, may be used to reduce the local recurrence rate. However, there is no consensus on the optimal use of curettage, along with fillers and adjuvants, to limit the recurrence rate. We performed a systematic review of articles using the terms long bones, GCTB, and treatment. Case reports, reviews, opinion articles, or technique notes were excluded based on the abstract. Twenty-six articles included in this review were then studied to establish the index in suggesting the surgical treatment of GCTB. The patient's gender, their age, the Campanacci grade of their tumor, and the type of surgery they had were not significantly associated with the local recurrence rate. Local recurrences seemed to be associated with the site of the tumor, occurring more frequently in the proximal femur or distal radius. A pathological fracture was not a contraindication for intralesional curettage. Treatment with denosumab did not decrease the local recurrence rate in patients who had been treated with curettage. The current literature seems to suggest that the ideal treatment for GCTB is to remove the tumor while preserving as much of the joint as possible. Local recurrent tumors can be treated with curettage to keep the re-recurrence rate within an acceptable limit. The choice for how to treat GCTB in the proximal femur or distal radius requires special attention.


A novel hybrid 3D endoscope zooming and repositioning system: Design and feasibility study.
Manipulation of the endoscope during minimally invasive surgery is a major source of inconvenience and discomfort. This report elucidates the architecture of a novel one-hand controlled endoscope positioning device and presents a practicability evaluation. Setup time and total surgery time, number and duration of the manipulations, side effects of three-dimensional (3D) imaging, and ergonomic complaints were assessed by three surgeons during cadaveric and in vivo porcine trials. Setup was accomplished in an average (SD) of 230 (120) seconds. The manipulation time was 3.87 (1.77) seconds for angular movements and 0.83 (0.24) seconds for zooming, with an average (SD) of 30.5 (16.3) manipulations per procedure. No side effects of 3D imaging or ergonomic complaints were reported. The integration of an active zoom into a passive endoscope holder delivers a convenient synergy between a human and a machine-controlled holding device. It is shown to be safe, simple, and intuitive to use and allows unrestrained autonomic control of the endoscope by the surgeon.


Two low-cost digital camera-based platforms for quantitative creatinine analysis in urine.
In clinical analysis creatinine is a routine biomarker for the assessment of renal and muscular dysfunctions. Although several techniques have been proposed for a fast and accurate quantification of creatinine in human serum or urine, most of them require expensive or complex apparatus, advanced sample preparation or skilled operators. To circumvent these issues, we propose two home-made platforms based on a CD Spectroscope (CDS) and Computer Screen Photo-assisted Technique (CSPT) for the rapid assessment of creatinine level in human urine. Both systems display a linear range (r(2) = 0.9967 and 0.9972, respectively) from 160 μmol L(-1) to 1.6 mmol L(-1) for standard creatinine solutions (n = 15) with respective detection limits of 89 μmol L(-1) and 111 μmol L(-1). Good repeatability was observed for intra-day (1.7-2.9%) and inter-day (3.6-6.5%) measurements evaluated on three consecutive days. The performance of CDS and CSPT was also validated in real human urine samples (n = 26) using capillary electrophoresis data as reference. Corresponding Partial Least-Squares (PLS) regression models provided for mean relative errors below 10% in creatinine quantification.


Current state and trends of access to sanitation in Ethiopia and the need to revise indicators to monitor progress in the Post-2015 era.
Investigating the current level and trends of access and identifying the underlying challenges to sanitation system development will be useful in determining directions developing countries are heading as they plan to promote sustainable development goals (post 2015 agenda). This research investigates the status and trends of access to improved sanitation coverage (ISC) in relation to the MDG target in Ethiopia with the aim of identifying prevailing constraints and suggesting the way forward in the post-MDG era. We examined data from a nationwide inventory conducted in accordance with the sanitation ladder at the national level and from a household survey in randomly selected urban slums in Addis Ababa. The inventory data were analyzed and interpreted using the conceptual model of the sanitation ladder. We used administrative reports and survey results to plot the time trend of the ISC. The data from the nationwide inventory of sanitation facilities, which are presented along the sanitation ladder reveal that more than half of the Ethiopian population (52.1%) still used unimproved sanitation facilities in 2014. The majority (35.6%) practiced open defecation, implying that the country is far from the MDG target for access to improved sanitation (56%). Most people in urban slums (88.6%) used unimproved sanitation facilities, indicating that the urban poor did not receive adequate sanitation services. Trend analysis shows that access to ISC has increased, but Central Statistical Authority (CSA) data reveal a decline. This discrepancy is due to differences in data collection methods and tools. Dry pit latrines are the most widely used toilet facilities in Ethiopia, accounting for about 97.5% of the ISC. The sanitation coverage is far from the MDG target and the majority of the population, mainly the urban poor, are living in a polluted environment, exposed to water and sanitation-related diseases. The sanitation coverage estimates might be even lower if proper utilization, regular emptying, and fecal sludge management (FSM) of dry pit latrines were considered as indicators. In order to enhance sanitation services for all in the post-MDG era, urgent action is required that will establish proper monitoring and evaluation systems that can measure real access to ISC.


The immunostimulatory activity and stability of grass carp (Ctenopharyngodon idellus) roe lectin.
A hexameric rhamnose-specific lectin with a molecular mass of 205kDa and exhibiting some N-terminal sequence similarity to other fish lectins was isolated from roe of the grass carp (Ctenopharyngodon idellus) by affinity chromatography on rhamnose-Sepharose and ion exchange chromatography by fast protein liquid chromatography on a Mono S column. The lectin exhibited mitogenic activity toward murine splenocytes with a potency lower than that of the plant lectin ConA. It exerted a stimulatory effect at a concentration of 10 micro g/ml on the phagocytic activity of seabream (Sparus sarba) macrophages. It was unstable toward heat (temperature > or =40 degrees C), acid (0.1M HCl), alkali (0.1M NaOH), trypsin and succinylation.


Cardiac magnetic resonance imaging: A teaching atlas with emphasizing current clinical indications.
Cardiovascular magnetic resonance (CMR) is an amazing technology that continues to provide new innovative approaches for evaluating the heart and blood vessels. It can assess cardiac morphology, function, perfusion, viability, coronary and peripheral arteries, and metabolism and tissue characterization. The basic pulse sequences of CMR include; Spin Echo, Gradient echo, and Steady stet free precision. Current clinical indications of CMR are multiple and continuously evolving. CMR often works in complementary fashions to other cardiac imaging techniques or to resolve residual diagnostic dilemma. The purpose of this illustrative review is to review current clinical applications of CMR and to provide physicians and technologists with simple, and regular CMR cases form daily practice. Each case discusses briefly the related clinical history, followed by CMR imaging findings, and simple discussion to highlight the role of CMR in a particular cardiovascular disorder.


Neonatal POCUS: Embracing our modern day "stethoscope".
The dramatic technologic advancements seen in ultrasound have accelerated the growth of point-of-care ultrasound (POCUS) in medicine. Neonatology has lagged behind other pediatric and adult specialties in incorporating POCUS into clinical practice despite there being numerous applications in cardiac and non-cardiac arenas. Widely available training programs are aiding in improving this situation but significantly more structure and orchestration for neonatal POCUS dissemination will be needed to fully actualize the potential for POCUS to augment its widespread clinical application.


Mercury content in museum and recent specimens of chiroptera in Japan.
The mercury content of insectivorous Chiroptera caught in 1890, when mercurial pesticides had not yet been in use, and preserved in a museum in alcohol were compared with those caught in 1965--1967 or 1970--1975, during and after the use of mercurial pesticides, and kept frozen. The mercury content of the Chiroptera caught in 1890 was lower than that of those caught during or after the use of mercurial pesticides. Difference in mercury content between bats caught during and after use was not significant.


Anticonvulsant therapy increases fentanyl requirements during anaesthesia for craniotomy.
This study was designed to determine whether patients receiving chronic anticonvulsant therapy demonstrate an altered requirement for fentanyl during anaesthesia. Sixty-one patients undergoing craniotomy were studied; 20 controls (MED = 0) who had never received anticonvulsants and 41 epileptics in whom therapeutic plasma concentrations of either one (MED = 1), two (MED = 2), or three (MED = 3) different anticonvulsants were documented. During anaesthesia with 60-70 per cent N2O in O2 and 0.2 per cent isoflurane, a maintenance dose (MD) of fentanyl was administered using a continuous variable-rate IV fentanyl infusion, supplemented by intermittent 50 micrograms IV boluses. In order to define the minimal dosage of fentanyl required, the MD was titrated according to increases or decreases in the heart rate and/or mean arterial pressure exceeding 15 per cent of baseline ward values. A progressively higher fentanyl MD was required in the epileptic patients (MED = 1-4.3 +/- 0.5 microgram.kg-1.hr-1; MED = 2-5.4 +/- 0.6; MED = 3-7.6 +/- 0.6) compared with the control MD (MED = 0-2.6 +/- 0.5) (P less than 0.001). These findings indicate that there appears to be a dose-effect relationship between the number of anticonvulsants received and the maintenance dose of fentanyl required during balanced anaesthesia.


Mutagenic and clastogenic effects of organic extracts from the Athenian drinking water.
Organic extracts of chlorinated drinking water collected from the water distribution system of Athens, Greece, were tested in parallel for their ability to cause mutations in the Salmonella His+ reversion system and sister chromatid exchanges (SCE) and chromosomal aberrations in CHO cells. They were found to induce His+ revertants in frequencies similar to those detected previously with drinking water extracts from other areas around the world. We observe that organic samples from surface drinking water induce SCE and chromatid type abnormalities in CHO cell cultures while samples collected from ground drinking water did not. These findings confirm that chlorinated surface drinking water contains substances which are mutagenic and clastogenic and may be carcinogenic in humans.


Genetic susceptibility to murine hepatocarcinogenesis is associated with high growth rate of NDEA-initiated hepatocytes.
The murine hybrids (C57BL/6J X C3Hf)F1 (B6C3) and (C57BL/6J X BALB/c)F1 (B6C), which have a high and low spontaneous and induced incidence of hepatocellular tumors, respectively, were treated with a single dose of NDEA at 1 week of age followed by TCPOBOP, a phenobarbital-like promoter of liver carcinogenesis, or by vehicle, and sacrificed at 30 weeks of age. The frequency per liver of hepatocellular nodules was similar in the two hybrids. However, in male mice the mean volume of nodules was about 10-fold greater in B6C3 than in B6C mice receiving NDEA followed by vehicle, and the treatment with TCPOBOP after NDEA stimulated nodule growth, with a much greater response in B6C3 mice. In female mice no differences in the mean volume of nodules were seen between hybrids after NDEA and vehicle, whereas upon NDEA and TCPOBOP treatment the mean volume of nodules was 25-fold greater in B6C3 than in B6C females. In addition, a few hepatocellular adenomas and carcinomas were observed, mostly in animals treated with NDEA and TCPOBOP, and they were 3-fold more numerous among B6C3 than B6C mice. TCPOBOP alone induced the same biochemical and hyperplastic effects in the liver of both hybrids. Using DNA probes homologous to Moloney murine leukemia virus, intracisternal A particle and virus-like 30S sequences, no correlation was apparent between the expression of any of these endogenous retroviral families and the strain susceptibility to hepatocarcinogenesis. We hypothesize that the different susceptibility to hepatocarcinogenesis between B6C3 and B6C mice is related to a higher growth rate of B6C3 than B6C initiated liver cells.


Miniaturization of a high-throughput pLDH-based Plasmodium falciparum growth inhibition assay for small volume samples from preclinical and clinical vaccine trials.
To date, no immune correlates for blood stage-specific immunity against Plasmodium falciparum malaria parasites have been identified. Growth and/or invasion inhibition assays using sera from Phase 2a/b trials will aid in determining whether correlations with protective immunity can be established for these assays. A major constraint in the ability to evaluate functional antibody activities from populations in endemic areas is the relatively limited availability of sufficient sample quantity. For this reason, we developed a miniaturized and high-throughput method to measure growth inhibitory activity by quantification of parasite lactate dehydrogenase (pLDH) in a 384-microtiter plate format. This culture method can be extended beyond the pLDH-based readout to other techniques commonly used to determine growth/invasion inhibition.


Goal-directed fluid therapy: stroke volume optimisation and cardiac dimensions in supine healthy humans.
Based on maximisation of cardiac stroke volume (SV), peri-operative individualised goal-directed fluid therapy improves patient outcome. It remains, however, unknown how fluid therapy by this strategy relates to filling of the heart during supine rest as reference for the anaesthetised patient and whether the heart becomes distended. To answer these questions, this study related SV to the diastolic filling of the heart while varying central blood volume (CBV) between hypo- and hyper-volaemia, simulating bleeding, and fluid loading, respectively, when exposing healthy human subjects to head-up (HUT) and head-down tilt (HDT). Twelve healthy volunteers underwent graded tilt from 20 degrees HDT to 30 degrees HUT. The end-diastolic dimensions of the heart were assessed by transthoracic echocardiography with independent evaluation of SV by Modelflow. The CBV was monitored by thoracic electrical admittance, central venous oxygenation and pressure, and arterial plasma atrial natriuretic peptide. Also, muscle and brain oxygenation were assessed by near infrared spectroscopy (n=7). The HUT reduced the mentioned indices of CBV, the end-diastolic dimensions of the heart, and SV. Conversely, HDT-enhanced tissue oxygenation and the diastolic filling of the heart, but not SV. In healthy supine humans, the heart is provided with a volume that is sufficient to secure a maximal SV without distending the heart. The implication for individualised goal-directed fluid therapy is that when a maximal SV is established for patients, cardiac pre-load is comparable to that of supine healthy subjects.


Immunochemical data suggesting a pattern for the evolution of human placental alkaline phosphatase.
Hyperimmune absorbed rabbit antisera which were reactive with epitopes specific for individual variants of human placental alkaline phosphatase were tested for their reactivity with primate placental alkaline phosphatases. Using the three epitope-specific reactivities defined previously, we found that: epitope I is present in the S-, D- and I-variants of human placental phosphatase, and in the chimpanzee and pygmy chimpanzee placentae; epitope II is present in the F- and 17-variants, and in the Nagao isoenzyme of human placental alkaline phosphatase, and in some orangutan placentae and all spider monkey placentae tested; epitope III is present in the F- and 17-variants, and the Nagao isoenzyme of human placental alkaline phosphatase, and in all the spider monkey placentae and the single squirrel monkey placenta examined. The binding assay was complemented by a competitive radioimmunoassay, which confirmed that the spider monkey placental samples were binding to the same antibody population which bound the human enzymes. The presence of epitopes characteristic of rare human placental phosphatase variants in these remote primate relatives suggests that the rare variants in the current human population have been present during the entire course of evolution. The presence of both epitopes characteristic of the Nagao isoenzyme in spider monkeys suggests that this variant isoenzyme is closely related to the enzyme present in the primate placenta at the time of species divergence (humans and New World monkeys). A hypothetical scheme for this divergence is proposed.


Removal of seeds from Neotropical frugivore droppings : Ant responses to seed number.
In a Costa Rican rain forest, the majority of tree and shrub species have their seeds dispersed by vertebrates. Over a third of the species' seeds are of a size accessible to, and primarily carried away by, ants. Frugivorous bird droppings come in sizes from a few seeds to over a thousand, with number of seeds isometric with bird body mass. Dropping size decreases as seeds are scattered by rain. Larger droppings produced from fruits ofMiconia affinis (Melastomataceae) are discovered and recruited to more by ants, with a diverse guild of 22 ant species carrying away the seeds. Droppings with 64 and 16 seeds were used by a subset of the ant community exploiting 4-seed droppings, likely due to resource defense by aggressive species. Seeds in the smallest droppings stood the smallest chance of being removed. Although soil-nesting ants of the tribe Attini were the primary removers of seeds in this forest, a third of removal was by ants living in ephemeral litter nests. Seeds ofM. affinis were found in litter ant nests in 8 of 28 1-m2 plots, suggesting that seed rain was not highly localized. Since litter nests are common, contain few seeds/nest and are often abandoned by their ants, litter ants may be the best candidates for ant-plant dispersal mutualisms.


Hard CiDRE revs up macrophage infectivity in COVID-19.
IL-10 has been linked to COVID-19 severity, but whether it is causal is unclear. In this issue of Immunity, Mitsui et al. find that a specific single-nucleotide polymorphism (SNP) results in expression of an IL10RB-IFNAR2 fusion protein that intensifies IL-10-mediated induction of ACE2 expression and viral infectivity of macrophages.


[Bone tumors and tumor-like lesions of the jaw. Findings from the Basel DOSAK reference registry].
Within this chapter, tumors which produce bone matrix (osteoid) or other bone-forming tissues, like cartilage, connective tissue, and cementum or originate within the bone marrow are described. In addition, those lesions are mentioned which, according to the WHO, are typical for the jawbones and are therefore included within the 1992 WHO classification of odontogenic tumors. Vascular and epithelial tumors which may also be observed in the jaws are not described. All the lesions and tumors reported have in common a more or less similar clinical and radiological appearance. A precise diagnosis, therefore, can only be established by a sufficient biopsy which should be seen by a pathologist with experience in this field. In addition to clinical, radiological, and histological appearances, therapeutic necessities are described. The data from recent publications are taken into account. This presentation is based on the data collected within the reference registry of the German-Austrian-Swiss Study Group of Tumors of the Face and Jaws (DOSAK) in Basel.


Association of Serum 25-Hydroxyvitamin D with Life Style and Dietary Factors in Egyptian Prepubescent Children.
There had been a growing evidence of high prevalence of vitamin D deficiency especially among children which may increase the risk of many chronic diseases in adulthood. Assessment of different lifestyles and dietary behaviour influencing the level of serum 25-OHD in a group of Egyptian prepubescent children. Two hundred boys and girls aged from 9 to 11 years were recruited from two primary public schools situated in Giza governorate in Egypt. A questionnaire was developed to obtain relevant information related to age, dietary habits, and physical activity. Thorough clinical examination and measurement of weight and height were performed. Body mass index was calculated. Serum samples were assayed for 25-hydroxy vitamin D (25-OHD). Low serum 25-OHD (< 20 ng/ml) was found in 11.5% of the whole sample. Mean serum 25-OHD was significantly lower in obese subjects and in those with low physical activity (p < 0.05). Multiple stepwise linear regression analysis showed that BMI and physical activity were the main predictors of serum 25-OHD (P < 0.05). Lifestyle factors in terms of physical activity and BMI may contribute significantly to the optimal vitamin D status of apparently healthy children.


Is HbA1c in the first trimester associated with adverse outcomes among pregnant Asian Indian women without gestational diabetes?
The aim of this study is to determine the association of elevated HbA1c in the first trimester (HbA1c-FT) with adverse events among pregnant Asian Indian women without gestational diabetes (GDM). This retrospective cohort study included 1618 pregnant women who delivered at a single urban tertiary care center and had HbA1c-FT estimation between January 2011 and September 2017. Those with GDM according to a 75-g OGTT after 24 gestational weeks were excluded. Multivariable logistic regression models assessed the association between elevated HbA1c-FT and adverse events. At a cutoff of ≥37 mmol/mol (5.5%), HbA1c-FT was associated with preterm birth at <37 gestational weeks (adjusted odds ratio (OR) 2.10, 95% CI 1.11-3.98). There was a continuum of risk for primary caesarean delivery with higher HbA1c-FT levels (adjusted OR per 5-mmol/mol (0.5%) increase in HbA1c-FT for primary caesarean delivery: 1.27, 95% CI 1.06-1.52). In the crude analysis, gestational hypertension was associated with HbA1c-FT, but not after adjustment for confounding factors. HbA1c-FT was not associated with other adverse events (macrosomia, large for gestational age babies, or other neonatal complications). Even without GDM, the results suggest an association of HbA1c-FT with preterm birth and primary caesarian delivery among Asian Indian women.