Penetration of titanium dioxide nanoparticles through slightly damaged skin in vitro and in vivo.
Titanium dioxide nanoparticles (TiO2-NPs) have been widely developed for versatile use, but the potential risk form their skin exposure is still unclear. To evaluate this risk, the skin penetration of TiO2-NPs is necessary to be understood first. The aims of this study are to investigated the penetration of TiO2-NPs through slightly damaged skin and intact skin in vitro and in vivo. TiO2-NPs with a diameter of 20 nm was labeled with 125I.The skin of rat was treated with 2% SLS solution and obtained as slightly damaged skin. The 125I labeled TiO2-NPs (125I-TiO2-NPs)solution and 0.9% PS solution were added into the donor chamber and receptor chamber of static diffusion cells which clamped the skin at the middle of two half-cells, respectively. During 24 hours, samples were extracted from the receptor chamber and counted for 1 min using γ-counter to detect the radioactivity. The skin penetration of TiO2-NPs in vitro was expressed as the percentage of radioactivity of receptor chamber solution compared with total radioactivity in the donor chamber. Thereafter, the 125I-TiO2-NPs was exposed to the rats. After 1 day and 3 days, the blood and tissues of rats were harvested, weighed and counted for 1 min using γ-counter to detect the tissue radioactivity. The skin penetration of TiO2-NPs in vivo was expressed as the percentage dose per gram tissue (% dose/g). In the skin penetration experiment in vitro, the radioactivity of receptor chamber solution through damaged skin was higher than that of through intact skin and was about 2% radioactivity of donor chamber on 24 h. In the skin penetration experiment in vivo, the radioactivity of blood and tissues of rats after exposing to 125I-TiO2-NPs solution though damaged skin or intact skin were less than 0.05% dose/g on 1 d and quickly declined on 3 d. The skin penetration rates of TiO2-NPs through slightly damaged skin and intact skin in vitro and vivo were lower than the rate of free 125I in the TiO2-NPs solution. The TiO2-NPs could not penetrate through the damaged skin or intact skin both in vitro and in vivo. It suggested that the TiO2-NPs should be safe when it was applied and contacted with skin.


Functional characterization of Fospeg, and its impact on cell cycle upon PDT of Huh7 hepatocellular carcinoma cell model.
Although several treatment options are available for hepatocellular carcinoma (HCC), their application is mostly restricted to early diagnosed cases or includes liver transplantation, which is rarely available due to donor scarcity. The attractiveness of PDT as a cancer treatment does not only come from its minimal invasiveness, but also from the high selectivity due to tumor localization that can be applied. Precise focusing of light on tumor lesions will result in tumor-specific PDT activation. Novel photosensitizers can be applied in such low concentrations that cells not subjected to irradiation remain healthy. The lethal effect and mechanism of death induction of the photosensitizer Fospeg has never been studied on hepatocellular carcinoma. The aim of the present study is to functionally analyze the impact of PDT on Huh-7 HCC cell line, as well as to analyze its impact on cell cycle protein expression. Cellular viability, and proliferation assays were conducted via MTT and BrdU assay, respectively. Transfected cell models of Huh7 with different constructs harboring cell cycle genes and downstream reporter luciferase gene were generated. Our results show a statistically significant decrease in both viability and proliferation of Huh-7 cells following PDT, while maintaining Fospeg and laser concentrations far below toxic levels. Proliferative cell cycle genes show a tendency of inhibition, while p53 levels show a significant increase following PDT. Fospeg-mediated PDT is a promising strategy for treatment of hepatocellular carcinoma and needs to be further explored in vivo.


Hepatitis B virus is inhibited by RNA interference in cell culture and in mice.
For chronic hepatitis B virus (HBV) infection the effects of current therapies are limited. Recently, RNA interference (RNAi) of virus-specific genes has emerged as a potential antiviral mechanism. Here we studied the effects of HBV-specific 21-bp short hairpin RNAs (shRNAs) targeted to the surface antigen (HBsAg) region and the core antigen (HBcAg) region both in a cell culture system and in a mouse model for HBV replication. HBsAg and hepatitis B e antigen (HBeAg) in the media of the cells and in the sera of the mice were analyzed by time-resolved immunofluorometric assay, intracellular HBcAg by immunofluorescence assay, HBsAg and HBcAg in the livers of the mice by immunohistochemical assay, HBV DNA by fluorogenic quantitative polymerase chain reaction (FQ-PCR) and HBV mRNA by semi-quantitative reverse transcriptase PCR (RT-PCR). Transfection with the shRNAs induced an RNAi response. Secreted HBsAg was reduced by >80% in cell culture and >90% in mouse serum, and HBeAg was also significantly inhibited. Immunofluorescence detection of intracellular HBcAg revealed 76% reduction. In the liver tissues by immunohistochemical detection, there were no HBsAg-positive cells and >70% reduction of HBcAg-positive cells for shRNA-1. And for shRNA-2 the detection of HBsAg and HBcAg also revealed substantial reduction. The shRNAs caused a significant inhibition in the levels of viral mRNA relative to the controls. HBV DNA was reduced by >40% for shRNA-1 and >60% for shRNA-2. RNAi is capable of inhibiting HBV replication and expression in vitro and in vivo and thus may constitute a new therapeutic strategy for HBV infection.


A berberine hydrochloride-carboxymethyl chitosan hydrogel protects against Staphylococcus aureus infection in a rat mastitis model.
Staphylococcus aureus (S. aureus) is the major pathogen responsible for mastitis in dairy cows, an important threat to their health, but prevention of S. aureus infection of the mammary gland remains challenging. Berberine hydrochloride (BH), a naturally occurring phytochemical, exhibits a wide range of activities, including antibacterial effects on S. aureus. In this study, we prepared a novel berberine hydrochloride-carboxymethyl chitosan hydrogel (BH-CMCH) with excellent thermosensitivity, injectability and in vitro antibacterial activity. In a rat model of mastitis induced by S. aureus, mammary duct injection of BH-CMCH reduced the bacterial load in infected mammary gland tissue and protected the tissue from damage from infection. In addition, proteomics analysis showed that mammary duct injection of BH-CMCH enhanced autolysosome degradation and promoted the innate immune response by activating the lysosomal pathway and up-regulating related significantly differentially expressed proteins (SDEPs). Taken together, the findings support the potential of BH-CMCH as an antibacterial agent against S. aureus-induced mastitis.


Effects of eccentric exercise-induced muscle injury on blood levels of platelet activating factor (PAF) and other inflammatory markers.
It has been reported that exercise with eccentric contractions can induce damage and inflammation in human muscle tissue, the severity of which depends on the duration and the intensity of exercise. Platelet activating factor (PAF) is a potent inflammatory mediator implicated in a series of pathophysiological conditions. We sought to investigate the relationship between PAF and eccentric exercise induced muscle damage. Thirteen healthy, recreationally active male subjects (27.5+/-3.78 year) performed 36 maximum voluntary eccentric contractions on a motorized muscle dynamometer using the elbow flexor muscles of the non-dominant arm. Venous blood samples were collected immediately before and after exercise at 2, 24, 48, 72 and 96 h. PAF was isolated, purified and determined by a platelet aggregation assay. Serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), C-reactive protein (CRP) and complement C3 (C3), plasma level of fibrinogen and whole blood level of leukocytes (and their subsets) were determined. Established indicators of muscle damage as maximum isometric torque (MIT), range of motion (ROM), relaxed arm angle (RANG), flexed arm angle (FANG), arm circumference (CIRC) and muscle soreness were also measured at the same time points. PAF, leukocytes, CK and LDH were elevated after exercise, while other biochemical parameters such as CRP, C3 and fibrinogen were unchanged. We also observed an inverse association between PAF and MIT and ROM, as well as a positive association with other markers of muscle injury, i.e. CK, LDH, FANG and CIRC (all P's<0.05). Our findings may imply a role for PAF in the mechanism of eccentric exercise induced muscle damage.


Alternating copolymerization of carbon dioxide and cyclohexene oxide catalyzed by salen CoIII(acetate) complexes.
A series of CoIII carboxylate based upon N,N,O,O-tetradentate Schiff base ligand framework have been prepared. X-ray diffraction analysis confirms that these Schiff base CoIII carboxylate are all monomeric species with a six-coordinated central Co in their solid structures. The activities and polycarbonate selectivity of these complexes toward the copolymerization of epoxide (cyclohexene oxide and propylene oxide) and carbon dioxide have been investigated in the presence of bis(triphenylphosphine)iminium chloride. Copolymerization experiments indicate that [bis(α-methyl-3,5-di-tertbutyl-salicylaldehyde) ethylenediiminato] CoIIIOOCH3 exhibits the highest activity and polycarbonate selectivity among these CoIII carboxylate. The resultant copolymer contained almost 100 % carbonate linkages with the molecular weight up to 71.8 kg mol-1 as well as narrow polymer dispersity index (polymer dispersity index = 1.5). The substituents and the mode of the bridging part between the two nitrogen atoms both exert significant influences upon the progress of the copolymerizations, influencing both the polycarbonate selectivity and the rate of copolymerization.


Phosphorylated testis-specific serine/threonine kinase 4 may phosphorylate Crem at Ser-117.
We aimed to investigate the internal existence status of testis-specific serine/threonine kinase 4 (Tssk4) and the interaction of Tssk4 and Cre-responsive element modulator (Crem). The internal existence status of Tssk4 in testis of mice was detected using western blotting and dephosphorylation method. The interaction of Tssk4 and Crem was analyzed by western blotting, immunohistochemistry, immunofluorescence, in vitro co-immunoprecipitation assays, and in vitro kinase assay. The results revealed that Tssk4 existed in testis both in phosphorylation and unphosphorylation status by a temporal manner with the development of testis. Immunofluorescence results showed that Tssk4 had identical distribution pattern with Crem in testis, which was utterly different to the localization of Cre-responsive element binding (Creb). In conclusion, our study demonstrated that phosphorylated Tssk4 might participate in testis genes expressions by phosphorylating Crem at Ser-117.


Incidental liver lesions seen on Breast MRI: When is additional imaging warranted?
Incidental hepatic lesions identified on breast MR can be a diagnostic dilemma due to concern for liver metastases or other significant hepatic lesions. The purpose of this study was to identify the incidence and nature of liver lesions seen on breast MR, and determine if additional imaging is necessary. Imaging reports of all breast MR examinations performed at our institution from January 1, 2010 to December 31, 2011 were reviewed to identify reports with hepatic abnormalities. Lesion characteristics, subsequent diagnosis, duration of follow up and additional imaging results (if performed) were all recorded. Of 1664 breast MRs, incidental hepatic lesions were seen in 207 studies (12.4%) in 169 patients. In 154 of 169 patients (91.1%) the lesions were characterized as T2 hyperintense and clearly as bright as adjacent fat on T2-weighted or localizer sequences. 0 of these 154 lesions were clinically significant at clinical or radiological follow-up. In the remaining 8.9% (15 of 169), lesions were characterized as not as bright as adjacent fat on T2 weighted or localizer imaging. In two cases, lesions were confirmed as incidental hepatic metastatic disease. 91.1% of incidental hepatic lesions were circumscribed, T2 hyperintense lesions and characterised as clearly as bright as adjacent fat on T2 weighted imaging at additional review. None of which were clinically significant at clinical or radiological follow-up. We advocate that circumscribed T2 hyperintense lesions which are clearly as bright as adjacent fat on T2 weighted imaging are of unlikely clinical significance and follow-up imaging should not be recommended, reducing the rate of additional imaging from 37.3% to 5.3%.


Cytochrome P450 oxidoreductase genetic polymorphisms A503V and rs2868177 do not significantly affect warfarin stable dosage in Han-Chinese patients with mechanical heart valve replacement.
To investigate the influence of cytochrome P450 oxidoreductase (POR) polymorphisms (A503V and rs2868177) on warfarin stable dosage (WSD) in Han-Chinese patients with mechanical heart valve replacement (MHVR). Three hundred and seventeen Han-Chinese MHVR patients on stable maintenance dose of warfarin were enrolled. Blood samples were collected for genotyping analyses of VKORC1 -1639G>A, CYP2C9 *3, CYP4F2 rs2108622 and POR (A503V and rs2868177). Average WSD of carriers with variant POR genotypes or haplotypes were compared. Association analyses were performed by single and multiple linear regression analysis. The variant allele frequencies of POR polymorphisms (A503V and rs2868177) were 38.8 % and 44.8 %, respectively. D' between POR A503V and rs2868177 was 0.855, r(2) was 0.375, and the frequencies of the four POR haplotypes were 42.3 % for CG, 36.3 % for TA, 18.9 % for CA, and 2.5 % for TG, respectively. There were no significant differences in average WSD among carriers with three variant POR A503V genotypes or among carriers with three variant POR rs2868177 genotypes (both P > 0.05). Similarly, there were no significant differences in average WSD among carriers with variant POR haplotypes (all P > 0.05). Neither single nor multiple linear regression analyses showed significant effects of POR A503V or POR rs2868177 polymorphisms on WSD. POR polymorphisms (A503V and rs2868177) do not appear to significantly influence WSD in Han-Chinese patients with MHVR.


The glutathione synthesis gene Gclm modulates amphiphilic polymer-coated CdSe/ZnS quantum dot-induced lung inflammation in mice.
Quantum dots (QDs) are unique semi-conductor fluorescent nanoparticles with potential uses in a variety of biomedical applications. However, concerns exist regarding their potential toxicity, specifically their capacity to induce oxidative stress and inflammation. In this study we synthesized CdSe/ZnS core/shell QDs with a tri-n-octylphosphine oxide, poly(maleic anhydride-alt-1-tetradecene) (TOPO-PMAT) coating and assessed their effects on lung inflammation in mice. Previously published in vitro data demonstrated these TOPO-PMAT QDs cause oxidative stress resulting in increased expression of antioxidant proteins, including heme oxygenase, and the glutathione (GSH) synthesis enzyme glutamate cysteine ligase (GCL). We therefore investigated the effects of these QDs in vivo in mice deficient in GSH synthesis (Gclm +/- and Gclm -/- mice). When mice were exposed via nasal instillation to a TOPO-PMAT QD dose of 6 µg cadmium (Cd) equivalents/kg body weight, neutrophil counts in bronchoalveolar lavage fluid (BALF) increased in both Gclm wild-type (+/+) and Gclm heterozygous (+/-) mice, whereas Gclm null (-/-) mice exhibited no such increase. Levels of the pro-inflammatory cytokines KC and TNFα increased in BALF from Gclm +/+ and +/- mice, but not from Gclm -/- mice. Analysis of lung Cd levels suggested that QDs were cleared more readily from the lungs of Gclm -/- mice. There was no change in matrix metalloproteinase (MMP) activity in any of the mice. However, there was a decrease in whole lung myeloperoxidase (MPO) content in Gclm -/- mice, regardless of treatment, relative to untreated Gclm +/+ mice. We conclude that in mice TOPO-PMAT QDs have in vivo pro-inflammatory properties, and the inflammatory response is dependent on GSH synthesis status. Because there is a common polymorphism in humans that influences GCLM expression, these findings imply that humans with reduced GSH synthesis capabilities may be more susceptible to the pro-inflammatory effects of QDs.


An elevated maternal plasma, but not amniotic fluid, soluble fms-like tyrosine kinase-1 (sFlt-1) at the time of mid-trimester genetic amniocentesis is a risk factor for preeclampsia.
The purpose of this study was to determine if an elevated concentration of soluble fms-like tyrosine kinase-1(sFlt-1) in maternal plasma and amniotic fluid is a risk factor for the subsequent development of preeclampsia. A case-control study was conducted to compare mid-trimester concentrations of maternal plasma and amniotic fluid sFlt-1 in patients who developed preeclampsia with those who did not. The study included 32 cases with preeclampsia (18 cases: severe preeclampsia) and 128 matched controls with normal outcomes. Patients with an abnormal fetal karyotype or major anomaly, multiple pregnancies, chronic hypertension, diabetes, and renal disease were excluded. Soluble Flt-1 concentration was measured by specific immunoassay. Nonparametric techniques were used for statistical analysis. 1) The median maternal plasma, but not amniotic fluid, sFlt-1 concentration in patients who developed preeclampsia was significantly higher than in the control cases (maternal plasma: median 730 pg/mL, range 60-3375 pg/mL vs median 441 pg/mL, range 58-1959 pg/mL, P < .05; amniotic fluid: median 10,504 pg/mL, range 5253-38,023 pg/mL vs median 10,236 pg/mL, range 4326-87,684 pg/mL, P = .65). 2) The median plasma concentration of sFlt-1 was higher in cases of severe preeclampsia than in those with mild preeclampsia without reaching statistical significance (median 762 pg/mL, range 261-3309 pg/mL vs median 334 pg/mL, range 60-3375 pg/mL; P = .07). However, there was no significant difference in the median amniotic fluid sFlt-1 concentrations between patients with severe preeclampsia and those with mild preeclampsia (P = .45). 3) An elevated maternal plasma sFlt-1 concentration (higher than 700 pg/mL) is a risk factor for the development of preeclampsia (OR 3.9, 95% CI 1.7-8.6) and severe preeclampsia (OR 7.4, 95% CI 2.5-22.1) after genetic amniocentesis. 4) The median interval from amniocentesis to the diagnosis of preeclampsia in patients with maternal plasma sFlt-1 concentrations higher than 700 pg/mL was 117 days (range 19-154 days). An elevated concentration of sFlt-1 in maternal plasma at the time of mid-trimester amniocentesis is a risk factor for the subsequent development of preeclampsia.


Oxidases responsible for resistance to pyrethroids sensitize Helicoverpa armigera (Hübner) to triazophos in West Africa.
Helicoverpa armigera (Hübner) is the major insect pest of cotton in Africa, Turkey, Asia, India, Indonesia and Australia. Populations recently developed resistance to pyrethroids in West Africa via the overproduction of cytochrome P450 (oxidases) increasing pyrethroid metabolism. One way to overcome pyrethroid resistance is to use compounds that show negative cross-resistance to pyrethroids. Triazophos is one of these compounds: it is slightly more toxic against pyrethroid resistant larvae of H. armigera than against susceptible ones. Overproduced oxidases transform the non active triazophos into its active form, triazophos-oxon, since this form was significantly more often found in larvae from pyrethroid resistant strain (23%) than in susceptible strain (15%). This suggests that oxidases, which provide resistance by degradation of pyrethroids in the resistant individuals, also activate triazophos in its toxic oxon form resulting in a negative cross-resistance.


Estimating trends in the effectiveness of seat belts in saving lives, 1975-1985.
We examine the effectiveness of seat-belt systems for preventing mortality in automobile crashes. In particular, we examine whether seat-belt effectiveness has been constant over this period and conclude that there has been a trend toward an increased effectiveness of seat-belt systems. The relative odds of death associated with wearing seat belts dropped from 0.80 in 1975 to 0.34 in 1985. This trend results from improvements in both the type and design of seat belts and from additional design changes, such as padded dashboards and collapsible steering columns, that protect occupants at the time of collision.


Spontaneous luteinizing hormone surge and cleavage of in vitro fertilized embryos.
The importance of monitoring luteinizing hormone (LH) secretion during gonadotropin stimulation remains controversial. In the present study, the authors evaluated the occurrence of spontaneous LH surges in 170 cycles stimulated by clomiphene citrate and human menopausal gonadotropin, and correlated the success rate of embryo cleavage to the time interval between the occurrence of the LH surge peak value and the time of human chorionic gonadotropin (hCG) administration. LH was quantitated from urine by an avidin-biotin enzyme immunoassay. The results indicated that a spontaneous LH surge occurred in 18% of the cycles. The number of oocytes recovered was not affected by the occurrence of a spontaneous LH surge. In 12% of all cases, the spontaneous LH surge occurred less than 12 hours before the administration of hCG, and in these cases embryo cleavage was not reduced. In 6% of all cases, the spontaneous LH surge occurred over 12 hours before hCG administration, and in these cases embryo cleavage was reduced significantly.


Functional analysis of cus operon promoter of Klebsiella pneumoniae using E. coli lacZ assay.
The transcriptional fusion of reporter lacZ gene with cusRS regulatory promoter of cus operon of Klebsiella pneumoniae enabled us to analyze the inductive effect of copper on promoter via lacZ assay. The stimulus response curve of promoter to a range of copper metal concentrations indicated a normal sigmoidal response profile with apparent Hill coefficient 1.0. There was a positive correlation of promoter response to the increasing concentration of copper in the medium. AC(50) value of copper was calculated to be 1mM, whereas the promoter response was exponential beyond 1mM and up to 2.5mM concentration. The promoter activity did not increase exponentially in copper concentration higher than 2.5mM. The promoter PcusRS requires two chromosomally encoded regulatory proteins, CusS and CusR, in trans for maximal in vitro activation. The PcusRS regulatory promoter sequence also contained regulatory -10 and -35 boxes along with CusR binding motif. The results supported the concept of cus operon regulation as an essential mechanism for maintaining the cellular homeostasis at very high (e.g. 3mM), and even toxic copper concentrations.


The effects of capsaicin pre-treatment on the responses of single neurones to sensory stimuli in the trigeminal nucleus caudalis of the rat: evidence against a role for substance P as the neurotransmitter serving thermal nociception.
Rats were systemically pre-treated with capsaicin either on the first day of life or at an age of 1 month. Both treatments were found to deplete substance P levels in the trigeminal nucleus caudalis (55.6% and 57.9% depletions, respectively). Extracellular single neurone recordings in the trigeminal nucleus caudalis revealed that neither type of capsaicin treatment greatly altered the proportions of neurones responding to non-noxious or noxious mechanical stimulation of the face. However, the proportion of mechanically-nociceptive neurones also responding to noxious thermal stimulation was greatly reduced in neonatally-treated, but not adult-treated rats. As both methods of capsaicin treatment caused similar depletions of substance P, it is concluded that this peptide may not be the neurotransmitter of afferent fibres to the trigeminal nucleus caudalis signalling thermal nociception.


Inactivated Four-Factor Prothrombin Complex Concentrate Dosing Practices for Reversal of Warfarin-Related Intracranial Hemorrhage.
Inactivated four-factor prothrombin complex concentrate (I4F-PCC, Kcentra®) has become an important agent for the urgent or emergent reversal of bleeding associated with vitamin K antagonists such as warfarin. There is recognized inter-institutional variability with the use of I4F-PCC, especially as it relates to dosing practices. We sought to characterize variations in I4F-PCC dosing practices and their impact on patient outcomes and describe overall real-world clinical practice surrounding I4F-PCC utilization in the context of the management of warfarin-related intracranial hemorrhage (ICH). This is a multicenter retrospective pragmatic registry study of adult patients admitted at a participating study site between January 1, 2014, and December 31, 2015, who received I4F-PCC for reversal of warfarin-related ICH. Practices around warfarin-related ICH reversal in context of I4F-PCC utilization are described, including repeat I4F-PCC dosing, adjunctive reversal agents, and dose rounding policies (i.e., rounding doses to nearest vial size vs preparing exact/unrounded doses). All research was approved by local human investigation committees at each institution. Seventeen institutions contributed data on 528 patients to this registry. These institutions were primarily urban centers (74%), located in the southeast USA (47%), with Level 1 Trauma designation (79%), and with Comprehensive Stroke Center designation (74%). Most patients included in the study had sustained a non-traumatic ICH (68%), had a median admission GCS of 14 (IQR 7-15), and were receiving warfarin for atrial fibrillation (57.4%). There was substantial time latency between baseline INR and I4F-PCC (median 2.4 h, IQR 1.4-4.5 h). Most patients received adjunctive reversal agents, including vitamin K (89.5%) and fresh frozen plasma (FFP) (31.9%). A smaller proportion (6.0%) of patients received repeat I4F-PCC dosing. The median ICU length of stay (LOS) was 3 days (IQR 2-7 days), median hospital LOS was 6 days (IQR 3-12 days), and overall mortality rate was 28.8%. For institutions rounding doses to the nearest vial size, the first post-I4F-PCC dose INR was statistically but not clinically significantly lower than for institutions without vial size dose rounding, with comparable degrees of INR reduction from baseline. No differences were observed between dose rounding cohorts in adverse effects, ICU or hospital LOS, modified Rankin score at discharge, or mortality rates. Most patients received single doses of I4F-PCC, with adjunctive reversal agents and rounding doses to vial size. The time difference from baseline INR to factor product administration is a potential opportunity for process improvement in the management of warfarin-related ICH.


Myeloprotection with activated carbon in doxorubicin-treated rats.
Chemotherapy can often cause a variety of side effects including bone marrow (BM) suppression, termed as myelosuppression. Accordingly, facile and effective management of chemotherapy-induced myelosuppression is currently a pivotal task for experimental pathologists and oncologists. Here, we chose to use activated carbon (AC) with an extensive surface area for studying its possible protective effectiveness with respect to BM in doxorubicin (DOX)-treated rats. Spherical AC with an extended surface area up to 4490 m2/g was prepared for per os (p/o) delivery, whereas for intraperitoneal (i/p) delivery we used the powdered form of AC that was derived from the aforementioned spherical AC. During the monthly treatment of animals with AC and DOX these two components were delivered alternately (not in the same day). After treatment, BM cells were isolated from femurs of sacrificed animals, stained with acridine orange (AO) and analyzed by flow cytometry. Regardless of the route of AC delivery (p/o or i/p), apparent myeloprotection with a possible regenerative effect was observed in animals that received DOX, as evidenced by recovery of the populations of total nucleated cells (TNC) and polychromatic (immature) erythrocytes accompanied by a considerable reduction of the number of apoptotic/dead cells among TNC (≤2.0%). Moreover, as a result of AC administrations, there was a significant increase of AO green and far-red fluorescence intensities in the population of TNC, which is suggestive of the ongoing quantitative and conformational changes in DNA and RNA associated with cell recovery and proliferation. Thus, AC preparations under the present experimental conditions can effectively tackle DOX-induced myelosuppression via mechanisms not necessarily associated with adsorptive detoxification.


Properly dividing with YAP.
The Hippo pathway prevents organ overgrowth and maintains tissue architecture by inhibiting the transcriptional coactivator YAP. In this issue of Science Signaling, Bui et al. find a role for YAP during cytokinesis that is independent of its transcriptional activity. This function of YAP may be important for maintaining genomic stability in dividing cells.


Outcome of Chagas cardiomyopathy in comparison to ischemic cardiomyopathy.
Chagas cardiomyopathy and ischemic heart disease (IHD) are frequent causes of chronic systolic heart failure (CHF) in areas where the former is endemic. Nonetheless, a specific comparison of outcome and role of etiology of CHF failure has not been performed in patients with both conditions. Two-hundred twenty two patients with Chagas cardiomyopathy and 79 with IHD with CHF were included in the study. A Cox proportional hazards model was used to establish independent predictors of mortality for the studied population. Survival analysis was performed with the Kaplan-Meir product limit method. In the multivariable model, Beta-Blocker therapy [(hazard ratio (HR)=0.36; 95% confidence interval (CI) 0.24 to 0.52; p<0.005)], Chagas etiology of CHF (HR=3.6; 95% CI 2.0 to 6.5; p<0.005), serum sodium levels (HR=0.95; 95% CI 0.91 to 0.98; p<0.005), digoxin use (HR=2.1; 95% CI 1.19 to 3.80, p=0.01), and spironolactone use (HR=1.7; 95% CI 1.10 to 2.80; p=0.02) were determined independent predictors of all-cause mortality for this cohort. Probability of survival at 12, 24, 36, 48, and 60 months was 92%, 92%, 88%, 81%, and 78%, respectively, in IHD patients, and 79%, 61%, 49%, 41%, and 35%, respectively, in Chagas cardiomyopathy patients (p<0.005). Outcome in patients with chronic systolic heart failure secondary to Chagas cardiomyopathy is poorer than that seen in those with IHD.


A statistical model for estimating donor postdonation platelet counts after plateletpheresis.
To avoid the need, in serial apheresis donors, either to delay plateletpheresis until a predonation platelet count is completed or to obtain a postdonation count after each procedure, a statistical model has been developed to predict the postdonation platelet count from the donor predonation platelet count, weight, and hematocrit. Predonation and postdonation platelet counts were measured in two groups of approximately 100 consecutive donors (Group A to test the model and Group B to validate it), and the postdonation counts were calculated with the model. Using stepwise multiple linear regression from donor data, estimated postdonation platelet counts were found to be comparable to the postdonation platelet counts actually measured. Estimated postdonation platelet counts x 10(9) per L (mean +/- SD) for each group, respectively, were Group A, 195 +/- 35, versus actual platelet counts of 195 +/- 39 (p = 0.43), and Group B, 183 +/- 36, versus actual platelet counts of 189 +/- 34 (p = 0.14). Sensitivity and specificity, respectively, were Group A, 57 and 99 percent and Group B, 62 and 99 percent. For most serial apheresis donors, application of this predictor model should preclude the need to obtain an extra postdonation platelet count.


Lentivector-mediated rescue from cerebellar ataxia in a mouse model of spinocerebellar ataxia.
Polyglutamine disorders are inherited neurodegenerative diseases caused by the accumulation of expanded polyglutamine protein (polyQ). Previously, we identified a new guanosine triphosphatase, CRAG, which facilitates the degradation of polyQ aggregates through the ubiquitin-proteasome pathway in cultured cells. Because expression of CRAG decreases in the adult brain, a reduced level of CRAG could underlie the onset of polyglutamine diseases. To examine the potential of CRAG expression for treating polyglutamine diseases, we generated model mice expressing polyQ predominantly in Purkinje cells. The model mice showed poor dendritic arborization of Purkinje cells, a markedly atrophied cerebellum and severe ataxia. Lentivector-mediated expression of CRAG in Purkinje cells of model mice extensively cleared polyQ aggregates and re-activated dendritic differentiation, resulting in a striking rescue from ataxia. Our in vivo data substantiate previous cell-culture-based results and extend further the usefulness of targeted delivery of CRAG as a gene therapy for polyglutamine diseases.


Tracheal and bronchial injury in high-frequency oscillatory ventilation and high-frequency flow interruption compared with conventional positive-pressure ventilation.
We compared the histopathologic changes in the airways of premature baboons treated with conventional positive-pressure ventilation (PPV) with those seen after high-frequency oscillatory ventilation (HFOV) and high-frequency flow interruption (HFFI). Twenty-six animals were treated with ventilation for 24 hours (five PPV, 10 HFOV, 11 HFFI), and 18 were treated with ventilation for 96 hours (six PPV, six HFOV, six HFFI). A semiquantitative scoring system was used to grade tissue changes in the trachea, carina, and both main-stem bronchi. Alterations were produced by all forms of mechanical ventilation. The degree of injury was similar and relatively mild for the PPV- and HFOV-treated animals at both 24 and 96 hours. Eleven of 17 baboons treated with HFFI ventilation (8/11 at 24 hours; 3/6 at 96 hours) had severe airway damage characterized by diffuse submucosal necrosis, extensive hemorrhage, dense polymorphonuclear leukocyte infiltration, sloughed epithelium, focal basophilia, and intraluminal debris. HFOV resulted in no greater degree of airway damage than did PPV. The use of HFFI, with the particular strategy we employed, resulted in a far greater degree of damage than either PPV (P less than 0.01) or HFOV.


A non-invasive method for estimating the severity of liver steatosis and the risk of fibrosis in non-obese type 2 diabetes patients with NAFLD.
Prognostic considerations include assessing the risk of liver fibrosis in people with non-alcoholic fatty liver disease (NAFLD). This study evaluates the use of hematologic and metabolic parameters regarding liver steatosis and fibrosis scores (FLI and Fib-4) in non-obese type 2 diabetes mellitus (t2DM) patients with NAFLD. Subjects underwent abdominal ultrasound examinations, and FLI and Fib-4 scores were calculated to evaluate liver steatosis and the risk of liver fibrosis non-invasively: 61 non-obese NAFLD subjects with t2DM were included in the cohort study and were divided into 2 groups depending on the t2DM treatment regimen. Fib-4 and WBC count demonstrated a significant inverse correlation (OR = 0.509, p = 0.007). WBC count had an R2 of 0.237, indicating that this marker could account for up to 23.7% of a variation in Fib-4. Fib-4 and FFA had positive correlation which did not achieve statistically significant prediction (OR=7.122, p=0.062). Additionally, a significant prediction of HbA1c (OR=1.536, p=0.016) and haemoglobin (OR=1.071, p=0.020) for FLI was revealed. HbA1c and other haematological and metabolic parameters, such as haemoglobin and WBC, may be another non-invasive tool for determining whether non-obese NAFLD patients with t2DM are at risk of developing liver steatosis and fibrosis.


Acute hyperglycaemia induces changes in the transcription levels of 4 major genes in human endothelial cells: macroarrays-based expression analysis.
Hyperglycaemia, in insulin-dependent or independent diabetes mellitus, promotes endothelial cell (EC) dysfunction and is a major factor in the development of macro- or microvascular diseases. The mechanisms and the disease-related genes in vascular diseases resulting from hyperglycaemia are poorly understood. Macroarrays. bearing a total of 588 cDNA known genes, were used to analyze HUVEC gene transcription subjected to 25 or 5-mM glucose for 24 h. Isolated mRNA derived from treated first passage HUVEC were reverse transcribed, 32P labeled, and hybridized to the cDNA macroarrays. Results show that acute hyperglycaemia induces an up-regulation of seven major genes, four of which were not previously reported in the literature. Northern blot analyses, performed on these 4 genes, confirm macroarrays results for alphav, beta4, c-myc, and MUC18. Moreover, time course analysis (0, 2, 4, 8, 2, 16, 24 h) of alphav, beta4 c-myc, and MUC18 mRNA expression, observed by northern blot assays, showed a peak at time points situated between 2 to 8 h. The 3 other genes (ICAM-1, beta1, and IL-8), were shown by others to be significantly upregulated after glucose stimulation. Furthermore, ELISA assays performed on the supernatant of HUVEC culture medium showed a significant increase of IL-8 for cells treated with 25-mM compared to 5-mM glucose. Identified genes, upregulated in endothelial cells as a result of acute hyperglycaemia, may serve as therapeutic or diagnostic targets in vascular lesions present in diabetic patients. These results also demonstrate the use of cDNA macroarrays as an effective approach in identifying genes implicated in a diseased cell.


An experimental analysis of treatment effects on consonant clusters and ambisyllabic consonants in two adults with developmental phonological problems.
The effect of treatment on errors in consonant clusters and in ambisyllabic consonants were investigated in two adults with histories of developmental phonological problems. Using a multiple baseline across behaviors, ambisyllabic consonant errors were treated in bisyllabic words and consonant cluster errors were treated in two different cluster classes. Results indicated that treatment, consisting of a sound-referenced rebus approach, affected change in production of trained words as well as generalization to untrained words for targeted behaviors.


Review of Mexican species of Podogaster Brullé (Hymenoptera: Ichneumonidae: Anomaloninae) with description of two new species.
Two new species of Podogaster Brullé, Podogaster brunneus n. sp. and Podogaster lagartensis n. sp., are described. The material was collected with Malaise traps operated for a year in the Ría Lagartos Biosphere Reserve, a dry tropical area of Southeast Mexico. Podogaster rosteri Gauld & Bradshaw is synonymized with Podogaster mexicanus (Cresson). A key to the Mexican species is also provided.


Immobilization of heavy metals in biochar by co-pyrolysis of sludge and CaSiO3.
Sludge pyrolysis has become an important method of sludge recycling. Stabilizing heavy metals in sludge is key to sludge recycling. Currently, research on the co-pyrolysis of sludge and industrial waste is limited. This study aims to explore the impact and mechanism of the co-pyrolysis of sludge and CaSiO3 (the main component of slag) and to achieve the concept of "treating waste with waste". To this end, we added different proportions of CaSiO3 (0%, 3%, 6%, 9%, 12%, and 15%) for the co-pyrolysis with sludge, and varied the pyrolysis temperatures (300, 400, 500, 600, and 700 °C) and retention times (15, 30, 60, and 120 min) to study heavy-metal stabilization in sludge. Consequently, the optimum dosage of CaSiO3 required for the immobilization of different heavy metals was 9% (Cu, Zn, Pb, and Cr) and 15% (Ni). The contents of Cu, Zn, Pb, Cr, and Ni in the stable state (oxidized and residual states) were 92.73%, 79.23%, 99.55%, 92.43% and 90.33% respectively. At a pyrolysis temperature of 700 °C, the steady-state proportions of Cr, Pb, and Zn were 88.12%, 90.21%, and 77.21%, respectively. At a pyrolysis temperature of 400 °C, the stable-Cu and -Ni contents were 97.21% and 99.43%, respectively. The optimal dwelling time was 15 min. The results showed that the CaSiO3 addition weakened the O-H stretching vibration peak intensity, promoted the formation of aromatic and epoxy ring structures, and enhanced the heavy-metal immobilization. Furthermore, the CaSiO3 decomposition during co-pyrolysis produced SiO2, CaO, and Ca(OH)2, which helped stabilize heavy metals.


Thymulin and the neuroendocrine system.
Thymulin is a thymic hormone exclusively produced by the thymic epithelial cells. It consists of a nonapeptide component coupled to the ion zinc, which confers biological activity to this molecule. After its discovery in the early 1970, thymulin was characterized as a thymic hormone involved in several aspects of intra- and extrathymic T-cell differentiation. Subsequently, it was demonstrated that thymulin production and secretion is strongly influenced by the neuroendocrine system. Conversely, an emerging core of information points to thymulin as a hypophysotropic peptide. Here we review the evidence supporting the hypothesis that thymulin is an important player in the hypophyso-thymic axis.


The role of risk aversion and brand-related factors in predicting consumers' willingness to buy expiration date-based priced perishable food products.
The objective of this study is to examine the relationship between risk-aversion, perceived quality, brand affect, brand trust, brand equity and willingness to buy expiration date-based priced perishable food products. Empirical data was collected from consumers with self-administrated questionnaires in Istanbul, Turkey. Hypotheses were developed based on the conceptual framework and were tested with structural equations modeling. Results indicate that risk aversion is positively related to perceived quality, brand affect, brand trust and brand equity. A positive relationship was also found between brand related-factors. Findings also demonstrated that risk aversion and brand-related factors have a positive effect on willingness to buy expiration date-based priced perishable food products.


Does Influenza Vaccination during Pregnancy Have Effects on Non-Influenza Infectious Morbidity? A Systematic Review and Meta-Analysis of Randomised Controlled Trials.
The recommendation to provide inactivated influenza vaccine (IIV) to pregnant women is based on observed protection against influenza-related morbidity in mother and infant. Non-live vaccines may have non-specific effects (NSEs), increasing the risk of non-targeted infections in females. We reviewed the evidence from available randomised controlled trials (RCTs) of IIV to pregnant women, to assess whether IIV may have NSEs. Four RCTs, all conducted in low- and middle-income settings, were identified. We extracted information on all-cause and infectious mortality and adverse events in women and their infants. We conducted meta-analyses providing risk ratios (RR). The meta-analysis for maternal all-cause mortality provided a RR of 1.48 (95% CI = 0.52-4.16). The estimates for miscarriage/stillbirth and infant all-cause mortality up to 6 months of age were 1.06 (0.78-1.44) and 1.11 (0.87-1.41), respectively. IIV was associated with a higher risk of non-influenza infectious adverse events, with meta-estimates of 2.01 (1.15-3.50) in women and 1.36 (1.12-1.67) in infants up to 6 months of age. Thus, following a pattern seen for other non-live vaccines, IIV was associated with a higher risk of non-influenza infectious adverse events. To ensure that scarce resources are used well, and no harm is inflicted, further RCTs are warranted.


The mediational role of physical activity, social contact and stroke on the association between age, education, employment and dementia in an Asian older adult population.
Our study aimed to investigate the pathways by which socio-demographic factors, modifiable health and lifestyle risk factors influence each other, and subsequently, lead to dementia. We used data from the Well-being of the Singapore Elderly study, a nationally representative survey of the older adult population aged 60 years and above in Singapore. Dementia diagnosis was established using 10/66 dementia criteria. Structural equation modelling (SEM) without latent variable was applied to confirm the hypothesized model. The results of SEM supported the hypothesized model (χ 2 = 14.999, df = 10, p = 0.132). The final model showed that those aged 75-84 years and 85 years and over (vs. 60-74 years), having no formal education, who had completed primary or secondary education (vs. completed tertiary), who were homemakers and retired (vs. paid work), and with a history of stroke were directly associated with higher odds of having dementia, while those who had more frequent contact with friends and neighbors as well as being physically active were directly associated with lower odds of having dementia diagnosis. The study also found that physical activity, more frequent contact with friends and stroke played a significant role as mediators in these relationships. The overall pathways model explained 57.7% of the variance in dementia. Our results suggest that physical activity, social contact and stroke were potential mediators in the relationship between age, education, employment and dementia. Intervention programmes focusing on physical activity such as exercise and social contact may be useful in reducing the risk of dementia among older adults.


Sirtuin 6 is a negative regulator of the anti-tumor function of natural killer cells in murine inflammatory colorectal cancer.
The immune system plays a crucial role in controlling colorectal cancer (CRC) development. Natural killer (NK) cells are tumoricidal but undergo exhaustion in CRC patients. The current research aims to understand the role of sirtuin 6 (SIRT6) in CRC-associated NK cell exhaustion in a murine inflammatory colorectal cancer model. To this end, inflammatory CRC was induced by treating mice with azoxymethane plus dextran sulfate sodium. The expression of SIRT6 in NK cells in murine mesenteric lymph nodes (mLNs) and the CRC tissue was characterized by Immunoblotting. SIRT6 knockdown was achieved by lentiviral transduction of murine splenic NK cells, followed by evaluation of NK cell proliferation and the expression of cytotoxic mediators using flow cytometry. NK cell cytotoxicity was measured by cytotoxicity assays. Adoptive transfer of murine NK cells was applied to analyze the effect of SIRT6 knockdown in vivo. We found that SIRT6 was up-regulated in infiltrating NK cells in the murine CRC tissue, especially NK cells with an exhausted phenotype and impaired cytotoxicity. SIRT6 knockdown significantly boosted murine splenic NK cell functionality, as evidenced by accelerated proliferation, increased production of cytotoxic mediators, and higher tumoricidal activity both in vitro and in vivo. Furthermore, the adoptive transfer of SIRT6-knockdown NK cells into CRC-bearing mice effectively suppressed CRC progression. Therefore, SIRT6 up-regulation is essential for murine NK cell exhaustion in CRC because it impedes the tumoricidal activity of murine NK cells. Artificial SIRT6 down-regulation could boost the function of infiltrating NK cells to oppress CRC progression in mice.


Two new fluorescent retrograde neuronal tracers which are transported over long distances.
Two new fluorescent retrograde neuronal tracers are reported: Nuclear Yellow (Hoechst S 769121), which mainly labels the neuronal nucleus; and Fast Blue (diamidino compound 253/50), which mainly labels the neuronal cytoplasm. Both tracers appear to be transported effectively over long distances in rat and cat.


Tight-binding streptavidin ligands from a cyclic peptide library.
During the screening of a soluble library of cyclo(AXXXXXAE)K-CONH2, a cyclic peptide cyclo(AHPQFPAE)K-CONH2 was identified as a tight-binding ligand (IC50 = 128 nM) and found to bind 1000-fold more tightly than its linear peptide to streptavidin. The results of this study suggest that library screening of conformationally constrained cyclic peptides can be an effective means for the discovery of high affinity ligands.


Early exposure to cannabis and risk for psychosis in young adolescents in Trinidad.
Cannabis use increases the risk for psychosis, but psychotogenic effects of cannabis may be restricted to exposure during early adolescence. Four hundred and seventy-two participants (aged 12-23 years), randomly selected from the general population in Trinidad, completed questionnaires on past and current cannabis use and psychotic symptoms (using the Community Assessment of Psychic Experiences). Cannabis use increased the risk of experiencing psychotic symptoms and this effect was conditional on early exposure, defined around the mean age of onset of cannabis use. Thus, exposure before but not after the age of 14 years predicted psychotic symptoms (respectively beta: 0.71, 95% CI 0.22; 1.19, P = 0.004 and beta: -0.11, 95% CI -0.57; 0.36, P = 0.66). The developmental effect of cannabis use was independent of use of other drugs or current use of cannabis. Early adolescence may be a critical period with regard to the psychotogenic effect of cannabis across geographical settings and ethnic groups.


The impact of new technologies in cervical cancer screening: results of the recruitment phase of a large randomised controlled trial from a public health perspective.
The decision to introduce liquid-based cytology (LBC) and HPV as screening tests involves criteria based on resource consumption. We used cross-sectional data at recruitment from the NTCC trial [ISRCTN81678807] on 28,000 women aged 35-60, randomised to receive a conventional Pap test or LBC plus HPV. We computed the resources employed to detect a CIN2+ with different screening strategies. In order to result in the same overall cost per CIN2+ detected as screening by conventional cytology, the unit cost of LBC used alone should be less than that of a conventional Pap while its unit cost may be up to 20% higher if HPV-triage for Atypical Squamous Cells of Undetermined Significance is applied together. With the same criterion the unit cost of HPV used alone may be about 20% higher than that of a Pap-test using a 1 pg/ml cut-off and over 40% higher using a 10 pg/ml cut-off. If HPV testing is applied with cytology-triage, a single HPV test may cost 20-30% more than a conventional Pap to result in the same overall cost per CIN2+ detected.


Natural Acidic Polysaccharide-Based Memristors for Transient Electronics: Highly Controllable Quantized Conductance for Integrated Memory and Nonvolatile Logic Applications.
As a leading candidate for further memory and computing applications, memristors are being developed in an important direction of transient electronics. Herein, wafer-scale acidic polysaccharide thin films are reported as promising materials for memristors with remarkable transient characteristics. The memristor shows freestanding and lightweight features, and can be fully dissolved in deionized water within 3.5 s. More importantly, the ion-confinement capability of acidic polysaccharides where the cations can interact with the ionizable acid groups enables atomic manipulation of conductive filament. As a result, (i) a single device can produce 16 highly controllable and independent quantized conductance (QC) states with quasi-nonvolatile and nonvolatile characteristics and (ii) QC switching can be performed with ultrafast speed (2-5 ns) and low energy consumption (0.6-16 pJ). These remarkable features make the memristor promising for fast, low-power, and high-density memory and computing applications. Based on QC switching, the encoding/decoding and nonvolatile basic Boolean logic are designed and implemented. More importantly, "stateful" material implication logic which is promising for future in-memory computing is demonstrated with QC switching. These results significantly advance acidic polysaccharides to develop nanodevices with quantum effects.


[Approaches to evaluation of occupational lead load in copper industry enterprise].
Evaluation of copper industry in Saxony revealed serum lead concentrations in 109 workers under the allowable level accepted in Germany for workers contacting lead, ambient lead concentrations in air of workplace rarely 2-4 times over the allowable level. The authors recommend these parameters of exposure as a basis for evaluating health state of workers.


COMT Val108/158Met genotype affects the mu-opioid receptor system in the human brain: evidence from ligand-binding, G-protein activation and preproenkephalin mRNA expression.
Recent data from [(11)C]carfentanil ligand-PET indicate that in the human brain, the availability of mu-opioid (MOP) receptor binding sites is affected by the Val(108/158)Met polymorphism of the catechol-O-methyltransferase (COMT) gene. This prompted us to validate the impact of COMT Val(108/158)Met on MOP receptors in human post-mortem brain. [(3)H]DAMGO receptor autoradiography was performed in frontal cortex, basal ganglia, thalamus and cerebellum (8 Met/Met, 6 Met/Val, 3 Val/Val). With respect to genotype, numbers of MOP binding sites in COMT Met(108/158) homozygous and Val(108/158)Met heterozygous cases were higher than in Val(108/158) homozygous. Differences were significant in the caudate nucleus (Val/Met vs. Val/Val), nucleus accumbens (Val/Met vs. Val/Val) and the mediodorsal nucleus of the thalamus (Met/Met vs. Val/Val). In the thalamus, this was corroborated by DAMGO-stimulated [(35)S]GTPgammaS autoradiography. Moreover, stepwise multiple regression taking into account various covariables allowed to confirm the COMT genotype as the most predictive factor in this structure. As a mechanism how COMT might exert its action on MOP receptors, it has been suggested that at least in striatopallidal circuits COMT Val(108/158)Met impacts on enkephalin, which is capable of reciprocally regulating MOP receptor expression. Thus, we assessed preproenkephalin mRNA by in situ hybridization. In the striatum, mRNA levels were significantly higher in COMT Met(108/158) homozygous cases indicating that MOP binding sites and enkephalin are regulated in parallel. Moreover, the transcript was not detectable in the thalamus. Thus, mechanisms other than an enkephalin-dependent receptor turnover must be responsible for COMT-related differences in MOP binding site availability in the human brain.


The use of the pectoralis major flap for advanced and recurrent head and neck malignancy in the medically compromised patient.
A retrospective review of seventy-one PPM flaps used between 1996 and 2010 primarily for oral and oropharyngeal squamous cell carcinoma presenting as either advanced stage IV primary disease (41/43), extensive recurrent (10) or metastatic (9) neck disease. The PPM flap was most commonly used following resection of the mandible (23) or the tongue/oropharynx (19). When the PPM flap was the preferred reconstruction option (54) the main indication, in addition to advanced disease, was significant medical co-morbidity (23). The majority of PPM flaps (75%) were used in the latter half of the series for an increasing number of patients in poor health with advanced disease. There was no evidence of an increase in age, ASA grade or extent of disease during this period. Approximately one quarter (17) of the flaps were used after failure of a free flap, most commonly a DCIA (7) or radial (6) flap. The 30day mortality in this group of compromised patients undergoing major surgery for advanced disease was 7% (5/71). The overwhelming majority had significant co-morbidity (94% grade 2 or higher with 63% ASA grade 3) and 90% had already undergone previous major surgery and/or radiotherapy. The 1-year, 3-year and 5-year overall survival rates were 65.5%, 39.1% and 11.0% respectively with cancer-specific survival rates of 82.0%, 65.5% and 65.5%. The majority died of disease related to the underlying co-morbidity. We recommend an aggressive approach to the surgical resection of advanced and recurrent disease but a pragmatic approach to reconstruction. The PPM major flap is reliable for reconstruction of defects of the mandible, tongue and oropharynx with a complete flap failure rate of 2.8%. Lateral defects of the mandible were managed without a plate and with an acceptable outcome in the context of limited life expectancy. This is the largest study of the use of the PPM flap for this type of patient group. The flap retains a major role in the management of advanced primary or recurrent disease, extensive metastatic neck disease and after failure of a free flap when in conjunction with significant co-morbidity.


Updates on Versatile Role of Putative Gasotransmitter Nitric Oxide: Culprit in Neurodegenerative Disease Pathology.
Nitric oxide (NO) is a versatile gasotransmitter that contributes in a range of physiological and pathological mechanims depending on its cellular levels. An appropriate concentration of NO is essentially required for cellular physiology; however, its increased level triggers pathological mechanisms like altered cellular redox regulation, functional impairment of mitochondrion, and modifications in cellular proteins and DNA. Its increased levels also exhibit post-translational modifications in protein through S-nitrosylation of their thiol amino acids, which critically affect the cellular physiology. Along with such modifications, NO could also nitrosylate the endoplasmic reticulum (ER)-membrane located sensors of ER stress, which subsequently affect the cellular protein degradation capacity and lead to aggregation of misfolded/unfolded proteins. Since protein aggregation is one of the pathological hallmarks of neurodegenerative disease, NO should be taken into account during development of disease therapies. In this Review, we shed light on the diverse role of NO in both cellular physiology and pathology and discussed its involvement in various pathological events in the context of neurodegenerative diseases.


[Interaction of calcium and some neuropharmacological agents on the isolated nerve fiber].
It has been suggested that calcium interferes with the effect of the local anesthetics (Blaustein and Goldman, 1966). Experiments were therefore made with the anionic phenobarbital and the cationic procaine and chlorpromazine in high (18 mM/1) and in low (0,18 mM/1) calcium concentration. These experiments showed that the effect of drugs used, and of changes in calcium concentration were independent of each other.


Late renal recovery after treatment over 1 year post-onset in an atypical hemolytic uremic syndrome: a case report.
Atypical hemolytic uremic syndrome (aHUS) is a life-threatening disease that leads to end-stage kidney disease if only a poor response to plasma exchanges (PEs) or eculizumab therapy is achieved. A 58-year-old Japanese man presented with thrombocytopenia, anemia, and kidney failure requiring dialysis without any underlying disease. A kidney biopsy revealed marked mesangiolysis in all glomeruli, compatible with thrombotic microangiopathy (TMA). Based on the positive anti- factor H antibody and negative result for secondary TMA, we diagnosed him as aHUS. Despite eculizumab administration after eight sessions of PE, neither platelet normalization nor kidney recovery was achieved. Eight months later, we discontinued eculizumab therapy due to anaphylactic reaction. At 15 months after the onset of TMA, his platelet count increased gradually from 40 to 150 × 103/μL with a decreased serum creatinine level and increased urine output, eventually allowing the withdrawal of dialysis therapy. A second kidney biopsy showed mesangial widening compatible with the healing of TMA. This case indicates that aHUS with PEs and eculizumab therapy has the potential for renal recovery even if over 1 year has passed.


Enzymatically Digested Food Waste Altered Fecal Microbiota But Not Meat Quality and Carcass Characteristics of Growing-Finishing Pigs.
This experiment aimed to evaluate meat quality, fatty acid profile in back-fat, and fecal microbiota of growing-finishing pigs fed with liquid enzymatically digested food waste. Fifty-six crossbred pigs (approximately 32.99 kg body weight) were assigned to one of two treatments with seven replicate pens and four pigs per pen. Pigs were fed with control (corn-soybean meal diets) or food waste from d 0 to 53, while all pigs were fed with the control diet from d 53 to 79. The 16S rRNA sequencing was used to analyze microbiota of feces collected on d 0, 28, 53, and 79. Meat quality and carcass characteristics were measured in one pig per pen at the end of the experiment. Pigs fed with food waste contained more (p < 0.05) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in back-fat. Feeding food waste increased (p < 0.05) the relative abundances of Lachnospiraceae and Ruminococcaceae, but decreased (p < 0.05) the relative abundances of Streptococcaceae and Clostridiaceae in feces on d 29 or d 53. In conclusion, feeding enzymatically digested food waste did not affect pork quality, but provided more beneficial fatty acids to pork consumers and altered the fecal microbiota in growing-finishing pigs.


Biochemical Diagnosis and Preoperative Imaging of Gastroenteropancreatic Neuroendocrine Tumors.
Neuroendocrine tumors are a group of neoplasms that can arise in a variety of locations throughout the body and often metastasize early. A patient's only chance for cure is surgical removal of the primary tumor and all associated metastases, although even when surgical cure is unlikely, patients can benefit from surgical debulking. A thorough preoperative workup will often require multiple clinical tests and imaging studies to locate the primary tumor, delineate the extent of the disease, and assess tumor functionality. This review discusses the biomarkers important for the diagnosis of these tumors and the imaging modalities needed.


Corticosteroids administered by nebulization to children with bronchial asthma.
Inhaled corticosteroids are recommended for long-term control of asthma in children, and nebulization simplifies administration to patients who lack hand-to-lung coordination. Information on the efficacy and safety of nebulized corticosteroids in children with mild to moderate asthma is limited, however, and comparison between corticosteroids is lacking. One hundred thirty-three patients 6 to 14 years of age with bronchial asthma were randomly assigned to receive flunisolide 500 microg or budesonide 500 microg, both administered twice daily by nebulizer for 4 weeks. Morning peak expiratory flow rate (primary efficacy endpoint) increased significantly from baseline (P<.001) with both medications (P = NS between treatments). Use of salbutamol as rescue medication decreased by 82.6% with flunisolide and by 82.7% with budesonide; respective decreases in asthma score were 78.8% and 82.3% (P<.001 vs baseline, P = NS between treatments). Only flunisolide reduced the number of nocturnal awakenings (P<.001). Ten patients in the flunisolide group and 12 in the budesonide group reported adverse events, none of which required discontinuation of treatment. In children with asthma, nebulized corticosteroids are effective and have good safety and tolerability profiles. Flunisolide and budesonide seem to produce equivalent clinical responses.


Histiocytosis-X: clonal culture, histocytochemistry, electron microscopy.
Pathologic interpretation of an osteolytic lesion from the skull of a 13-month-old boy was amplified by histocytochemistry of cells grown in a methylcellulose clonal culture system. Electron microscopy demonstrated the presence of X granules in the cytoplasm of malignant histiocytes, confirming a diagnosis of histiocytosis-X. Freshly fixed tissue containing histiocytosis-X cells and granulocytes showed histiocytosis-X cells that were positive for alpha-naphthyl acetate esterase (non-specific esterase) and negative with naphthol AS-D chloracetate as the esterase substrate (specific esterase). Clonal cell aggregates, harvested after 6 days' growth in culture, showed histiocytosis-X cells that were positive for both the nonspecific and specific esterases. Differences in staining reactions for the histiocytosis-X cells may be explained on the basis of immaturity of the histiocytosis-X cells growing in culture. This interpretation would support their origin from monocytes, monocytic precursors, or a still less differentiated myelomonocytic precursor cell. Furthermore, gel systems of clonally cultured cells appear to provide a useful tool for the growth and analysis of histiocytosis-X cells.


Schistosoma mansoni: species-selective response to N-chloroethyl-acetylcholine derivatives.
Three new acetylcholine mustard analogs were tested on schistosome and vertebrate neuromuscular preparations. These compounds extend a previously reported structure-activity series. The new compounds investigated include isopropyl-, cyclohexyl-, and benzyl-2-acetoxyethyl-2'-chloroethylamine (PrM, ChM, and BzM). In schistosome motor activity studies, all of the compounds caused irreversible reductions in the activity of Schistosoma mansoni after 1 hr exposure followed by 19 hr in drug-free medium. Under nonlethal conditions of dosage and exposure time, all compounds blocked carbachol-induced paralysis, indicating a possible action at schistosome cholinergic sites. All the compounds also reduced the labeling of schistosomes by dimethylaminonapthalene-5-sulfonamidoethyl dimethylamine hydrochloride (DDNS), a fluorescent ACh analog. In isolated vertebrate tissue experiments. PrM and ChM had slight agonist activity in the guinea pig ileum, and only PrM had agonist activity in the frog rectus abdominis preparation. PrM and BzM were found to antagonize ACh responses in the guinea pig ileum. Exposure of vertebrate tissues for 1 hr to high concentrations of ChM caused no long-lasting inhibition of ACh, pilocarpine, and serotonin (5HT) responses. Histamine responses were slightly reduced from control. Following 1 hr exposure of vertebrate tissues to PrM, initial reductions in all responses were seen, followed by recoveries to near control. BzM treatment, however, reduced all responses far below control; the tissues did not recover even after washing for 2 hr. It is concluded that ChM and, to a lesser extent, PrM, had a greater effect on schistosomes than on vertebrate tissues. BzM did not display species selectivity in this favorable direction.


Persistent inflammatory pain alters sexually-motivated behavior in male rats.
Urine from pro-œstrus female rodents evokes increased levels of sexually-motivated behaviors in males, including sniffing and scent marking of the urine spot as well as activation of brain reward regions. Stressors such as social defeat can adversely impact urine scent marking behavior in male rodents, an effect that can be mitigated with anti-depressant drugs. Persistent pain is also known to be a potent stressor, producing elevated levels of plasma corticosterone as well as reduced sucrose preference and reduced social interaction. However, the effect of persistent pain on sexually-motivated behavior is unknown. Here, we compared urine scent marking behavior in male rats for up to 3 weeks following intra-articular injection of Complete Freund's Adjuvant (CFA) or sham injection. CFA-injected rats exhibited profound and ongoing deficits in static weight bearing capacity. CFA-induced persistent inflammatory pain increased plasma corticosterone levels and reduced urine scent marking behavior in male rats. Moreover, while the vast majority of injured rats showed decreased urine scent marking preference for the pro-œstrus female urine spot, male rats with higher baseline scent marking preference also exhibited higher post-injury scent marking preference, more sniffing behavior and lower levels of plasma corticosterone, compared to those with lower baseline scent marking preference. Overall, scent marking behavior may be an ethologically relevant behavioral predictor of persistent pain-induced stress in rats, representing a novel translational approach to understanding chronic pain comorbidities.


Global mapping of cholera Vibrio and outbreaks in the Pre-Millennium Development Goals (MDG)/Sustainable Development Goals (SDG) and MDGs/SDGs era of 1990-2019.
Vibrio species and cholera outbreak yet remain a frequent health emergency despite progress made in integrated implementation of the MDGs/SDGs/WASH worldwide. Hence, this study aimed at appraising the impacts of MDGs/SDGs/WASH campaigns on the mitigation of cholera outbreak and associated consequences. The study mapped scientific production related to Vibrio outbreak from 1990 to 2019, identified trend, institutional/international concerted efforts toward outbreak research/response and gaps for future preparedness. Relevant documents were identified from the Web of Science database using an optimised title-field specific search Boolean that accommodated all pre-set inclusion criteria for the study. A total of 901 documents were identified including 869 available abstracts were retrieved for content-review of human incidence cases, mortality, culprit Vibrio species, strains, and biotypes. Explanatory analysis showed that the trend of outbreak documents approximately increased in 6th order quadratic relationship (R2 = 0.7948) from 1990 to 2019 with an annual growth rate of 3.21% and a mean value of 30.0 ± 18.0 per year. Other details revealed an increased and undulating case report/mortality rate of cholera outbreaks especially in the MDGs/SDGs era. Decadal comparison of Vibrio outbreak during the period showed significant variation in documents distribution (Kruskal-Wallis p = 0.00077). based on countries' efforts, the USA, ranked first in terms of article numbers (191), publication frequency (24.6(%) and total citations (5962). Four prevailing conceptual frameworks were identified in the outbreak documents with global community interest revealed as the largest topical coverage. All conceptual frameworks consisted in Vibrio characterisation, methodology-related, intervention-related, geographic-related concepts and some replete with health and climate-change depicting concepts. Also, the study observed high mortality in Vibrio outbreaks during 1990-1999 (29080 deaths), and 2010-2019 (386606 deaths) compared to 2000-2009 (7705 deaths) (Kruskal-Wallis, p < 0.05). High number of outbreaks due to V. cholerae and V. parahaemolyticus and a limited outbreaks attributed to emerging strains. In conclusion, vibrio outbreak has not lived up to various investment put into its control from various programme evolutions. The broad spectrum Vibrio vaccines that could cater for outbreak caused by common and emerging strains is inevitable and a significant thrust for future research.


Digestion and metabolism of high-tannin Eucalyptus foliage by the brushtail possum (Trichosurus vulpecula) (Marsupialia: Phalangeridae).
The digestion and metabolism of Eucalyptus melliodora foliage was studied in captive brushtail possums (Trichosurus vulpecula). The foliage was low in nitrogen and silica but high in lignified fibre and phenolics compared with diets consumed by most other herbivores. The high lignin content was suggested as the main cause of the low digestibility of E. melliodora cell walls (24%); microscopic observations of plant fragments in the caecum and faeces revealed few bacteria attached to lignified tissues. The conversion of digestible energy (0.34 MJ X kg-0.75 X d-1) to metabolizable energy (0.26 MJ X kg-0.75 X d-1) was low compared to most other herbivores, probably because of excretion of metabolites of leaf essential oils and phenolics in the urine. When the inhibitory effect of leaf tannins on fibre digestion was blocked by supplementing the animals with polyethylene glycol (PEG), intake of dry matter, metabolizable energy and digestible fibre increased. These effects were attributed to the reversal by PEG of tannin-microbial enzyme complexes. It was concluded that the gut-filling effect of a bulk of indigestible fibre is a major reason why the brushtail possum does not feed exclusively on Eucalyptus foliage in the wild.


Tenofovir alafenamide as compared to tenofovir disoproxil fumarate in the management of chronic hepatitis B with recent trends in patient demographics.
Tenofovir alafenamide (TAF) has recently been approved for chronic hepatitis B (CHB). It is more stable than tenofovir disoproxil fumarate (TDF) in the plasma and can provide similar efficacy with lower circulating concentration in patients with hepatitis B virus (HBV) infection. Areas covered: This synopsis will review the current anti-HBV standard practice and the changing epidemiology of CHB, specifically the controversies surrounding the renal and bone safety associated with TDF use in the context of an aging CHB population. We will review data from phase 3 registration trials, which demonstrated TAF was not inferior to TDF in antiviral efficacy for both HBeAg-positive and HBeAg-negative patients, while associated with less reduction in the estimated glomerular filtration rate and bone mineral density. Expert commentary: Current data supports the use of TAF as one of the first-line antiviral agents for general CHB patients without hepatic decompensation. However, more real-world data with long-term observation are needed to better define the role of TAF among other oral regimens. Additional studies are also needed to evaluate the efficacy and safety of TAF in special populations such as those with impaired hepatic function, existing impaired renal and/or bone function, and in pregnant women.


Mechanism of inhibition of human immunodeficiency virus type 1 reverse transcriptase and human DNA polymerases alpha, beta, and gamma by the 5'-triphosphates of carbovir, 3'-azido-3'-deoxythymidine, 2',3'-dideoxyguanosine and 3'-deoxythymidine. A novel RNA template for the evaluation of antiretroviral drugs.
Carbovir (the carbocyclic analog of 2'-3'-didehydro-2',3'-dideoxyguanosine) is a potent inhibitor of human immunodeficiency virus type 1 (HIV-1) replication. Assays were developed to assess the mechanism of inhibition by the 5'-triphosphate of carbovir of HIV-1 reverse transcriptase using either RNA or DNA templates that contain all four natural nucleotides. Carbovir-TP was a potent inhibitor of HIV-1 reverse transcriptase using either template with Ki values similar to that observed by AZT-TP, ddGTP, and ddTTP. The kinetic constants for incorporation of these nucleotide analogs into DNA by HIV-1 reverse transcriptase using either template were similar to the values seen for their respective natural nucleotides. In addition, the incorporation of either carbovir-TP or AZT-TP in the presence of dGTP or dTTP, respectively, indicated that the mechanism of inhibition by these two nucleotide analogs was due to their incorporation into the DNA resulting in chain termination. Carbovir-TP was not a potent inhibitor of DNA polymerase alpha, beta, or gamma, or DNA primase. Given the potent activity of carbovir-TP against HIV-1 reverse transcriptase and its lack of activity against human DNA polymerases, we believe that further evaluation of this compound as a potential drug for the treatment of HIV-1 infection is warranted.


Additive prognostic value of preoperative plasma glucose concentrations in calves undergoing abdominal surgery.
Surgical abdominal emergencies in calves are associated with a guarded prognosis, especially if neonates are affected. Because hypoglycemia has been associated with sepsis and endotoxemia, this study aimed to assess the prognostic relevance of preoperative plasma glucose concentrations (GLUC) in calves requiring surgery for an acute abdominal disorder. For this purpose, data from retrospective and prospective case series were analyzed, consisting of 586 and 83 hospitalized calves, respectively. The outcomes of calves were evaluated until hospital discharge (both study populations) and for 3 mo following discharge by a phone call to the farmer (prospective study population). For the retrospective study population, the overall survival rate was 31.2%. Calves with a negative outcome (NO) had significantly lower median GLUC (4.3 mmol/L) than calves with a positive outcome (PO; 5.0 mmol/L). The survival rates of calves with GLUC <2.4 mmol/L and 2.4 to 3.1 mmol/L were 3.6 and 8.3%, respectively. The inclusion of GLUC improved a previous prognostic model based on plasma l-lactate concentration and age. The resulting analyses indicated that NO was associated with low age (<7 d), hyper-l-lactatemia (>8.84 mmol/L), and GLUC <4.4 mmol/L (age 7-20 d) and <3.3 mmol/L (age ≥21 d), respectively. The area under the receiver operating characteristic curve of this model was 0.79 (95% confidence interval: 0.76-0.83) and the resulting sensitivity and specificity for NO at the optimal probability cut-point of 0.69 were 66.7 and 85.8%, respectively. For the prospective study population, the established model had sensitivity and specificity for predicting NO after 3 mo (proportion 24%) of 61.9 and 85%, respectively. In both study populations, hypoglycemia was significantly associated with intraoperative evidence of a septic process within the abdominal cavity. The present analyses show that hypoglycemia was highly indicative of a poor prognosis and serious intraoperative findings such as peritonitis. Determination of GLUC should therefore be part of the diagnostic work-up in calves suffering from an acute abdominal emergency.


Screening and regulatory network analysis of survival-related genes of patients with colorectal cancer.
The purpose of this study was to screen key survival-related genes from patients with colorectal cancer and explore signal transduction network of the involved genes. In a previous study, survival-related genes of patients with colorectal cancer were selected by colorectal cancer-related expression data GSE17538 using the Significance Analysis of Microarrays (SAM3.01) software, and 235 genes related to the survival of patients with colorectal cancer were obtained. Therefore, the following screening and analysis were conducted on these 235 genes in this study. First, the enrichment analysis of transcription factor binding sites was conducted on the 235 genes. Genes with more than seven transcription factor binding sites were screened. Then, these genes and upregulated genes in colorectal cancer were intersected. Finally, survival analysis and regulatory network analysis were conducted on the screened genes. This allowed clarification of the relationship between these genes and the survival of patients with colorectal cancer and the signaling network involving these genes in the cell signal transduction network of colorectal cancer. Through the above analysis, six upregulated genes in colorectal cancer related to the survival of colorectal cancer patients and highly regulated by transcription factors were selected, namely STX2, PODXL, KLK6, GRB10, EHBP1 and CREB5. These genes are involved in signal regulatory networks related to colorectal cancer metastasis-related signaling pathways. Therefore, the survival of patients with colorectal cancer is closely correlated with colorectal cancer metastasis. The six survival-related genes affect the survival of patients by regulating colorectal cancer metastasis-associated signaling pathways.


Site selection by Microphallus pygmaeus Levinsen, 1881 (Trematoda: Microphallidae) in the laboratory mouse.
When Microphallus pygmaeus Levinsen, 1881 was inoculated orally to laboratory mice, most specimens, failed to attach to the intestinal wall and passed out rapidly with the faeces. Those which did settle were distributed more or less evenly throughout the small intestine by 4 h after inoculation. By day 1 after inoculation the worms had developed to oviposition and most were found in the ileum. By 2 days after inoculation all worms were established, and they remained in this region where they grew and continued egg production with very little further mortality until day 6 after inoculation. Thereafter, gravid worms progressively detached and passed rapidly through the caecum, colon, and rectum with the faeces. All parasites had been voided by about day 12 inoculation. In spite of high mortality, distribution in the small intestine is also affected by the site of implantation.


Effects of omega-3 polyunsaturated fatty acids on plasma indices of thrombogenesis and inflammation in patients post-myocardial infarction.
To determine the effects of n-3 PUFAs supplementation on plasma indices of coagulation (fibrinogen), fibrin D-Dimer (an index of thrombogenesis and fibrin turnover), endothelial damage/dysfunction (von Willebrand factor (vWf)), platelet activation (soluble P-selectin (sP-sel)) and inflammation (interleukin-6, IL-6) in patients following acute myocardial infarction. Open-labelled randomised controlled trial. Seventy-seven post-myocardial infarction (MI) patients stabilized on standard secondary prevention therapy were randomised either to 3 months' treatment with Omacor 1 g/day (n=37) or 'usual care' control (n=40). Plasma levels of fibrinogen, D-Dimer, vWf, sP-sel, IL-6 and plasma viscosity at baseline and after 3 months were determined. At baseline, there were no significant differences between the groups in all research indices, except vWf levels were higher in patients allocated to Omacor supplementation. After 3 months, there were no significant changes in the levels of any research indices in either the Omacor supplemented or the 'usual care' control patients when compared to baseline. Patients who received Omacor experienced a fall in total cholesterol (p=0.019), total/HDL-cholesterol ratio (p=0.009) and LDL-cholesterol (p=0.023). However, the relative changes in plasma lipids and lipoproteins did not differ between the two groups. Three-month supplementation of Omacor at 1 g per day in post-MI patients is not associated with an improvement in the levels of peripheral indices of coagulation potential, endothelial function, platelet reactivity and inflammation.


Indications, safety, and feasibility of conversion of failed bariatric surgery to Roux-en-Y gastric bypass: a retrospective comparative study with primary laparoscopic Roux-en-Y gastric bypass.
Roux-en-Y gastric bypass (RYGB) is considered the "gold standard" revision procedure. The purpose of this study was to compare the surgical outcome of primary laparoscopic RYGB (pLRYGB) to revisional open or laparoscopic Roux-en-Y gastric bypass surgery (rRYGB). A retrospective analysis of all patients who underwent pLRYGB or rRYGB from January 2003 to December 2009 has been performed. Demographics, indications for revision, and complications have been reviewed. The rRYGB and pLRYGB patients have been compared. Seventy-two patients underwent rRYGB, and 652 patients underwent pLRYGB. Mean follow-up was 35 and 45 months, respectively. Fifty-six rRYGB procedures were performed laparoscopically. The primary operations had consisted of laparoscopic gastric banding (n = 28), laparoscopic vertical banded gastroplasty (n = 19), laparoscopic sleeve gastrectomy (n = 6), laparoscopic RYGB (n = 3), and biliopancreatic diversion with duodenal switch (n = 16). Indications included weight regain (n = 29), malabsorption (n = 16), gastrogastric fistula (n = 5), band-associated problems (n = 3), and refractory stomal ulceration (n = 1). There was no significant difference in early or late postoperative complications when comparing rRYGB to pLRYGBP patients (11.1% vs. 5.52%, P = 0.069 and 19.4% vs. 24.2%, P = 0.465 respectively). Five rRYGB patients (7.04%) required reintervention (3 internal hernias, 1 ventral hernia, 1 laparoscopic exploration) compared with 101 pLRYGB patients (15.71%; P = 0.051). None of the patients died. Mean hospital stay was not significantly longer in the rRYGB group (5.38 vs. 4.95 days, P = 0.058). In our series, hospital stay, morbidity, and mortality of rRYGB were not significantly higher compared with pLRYGB. Furthermore, we believe that this type of revisional bariatric surgery should be performed in high-volume bariatric centers.


Pelviureteric junction obstruction and recurrent abdominal pain in childhood.
Three cases of recurrent abdominal pain due to pelviureteric junction obstruction in childhood are presented. Ultrasound is useful in confirming the diagnosis and pyeloplasty is recommended.


Dynamical cell assemblies in the rat auditory cortex in a reaction-time task.
Simultaneous single unit spike trains were recorded in the auditory cortex of freely moving rats performing a complex cognitive task. The experimental paradigm is based on a two-choice task (Go/Nogo) with a two-component (pitch and location) auditory stimulus lasting 500 ms. We report evidence that firstly functional interactions, measured by cross-correlation analysis, between single units in the auditory cortex are dynamically modified in the period preceding the onset of the auditory stimulation, referred to as the 'waiting period'. We secondly observed that spatio-temporal firing patterns both within, and across cell spike trains also tended to appear in the waiting period, several seconds before the actual stimulus delivery. These patterns indicate a very precise repetition of spike discharges separated by long intervals (up to several hundreds of milliseconds). No consistent changes in mean rate were observed. These results suggest that network activity in the auditory cortex is selectively modified in rate independent ways before the actual sensory stimulation. These modifications may reflect participation of recurrent neuronal networks in processes anticipating the expected sensory input.


[Unusual presentation of a severe Mycoplasma pneumoniae infection: report of 2 cases].
Mycoplasma pneumonia is responsible for multisystemic infection. Pulmonary symptoms are most common in children. We describe herein two unusual severe forms of M. pneumoniae infection without initial pulmonary symptoms. The first case is an 8-month-old boy who was hospitalized in the pediatric intensive care unit with severe sepsis. There were no initial pulmonary symptoms, nor obvious clinical infection. Initial blood tests and x-ray did not aid the diagnosis. The blood tests came back positive for M. pneumonia. Pulmonary symptoms eventually appeared 24h later, and there was a pneumonia outbreak on the chest radiograph. The boy was given josamycin and improved quickly. The second case concerns an 8-year-old child who was hospitalized in the pediatric intensive care unit with toxic shock. No clinical infectious origin was found. A broad-spectrum antibiotic therapy was started with ceftriaxone and josamycin. The M. pneumoniae blood test came back positive, which confirmed the diagnosis of septic shock in M. pneumoniae, requiring adjustment of the antibiotic therapy. Current guidelines for the choice of probabilistic antibiotic therapy in case of severe sepsis do not include the case of M. pneumoniae. The early initiation of antibiotic therapy plays a major role in the prognosis of these patients. It seems useful to search for M. pneumoniae in cases of severe atypical infections, particularly in the absence of pulmonary symptoms.


Expressive genomic hybridisation: gene expression profiling at the cytogenetic level.
To describe a cytogenetic technique suitable for the rapid assessment of global gene expression that is based on comparative genomic hybridisation (CGH), and to use it to understand the relation between genetic amplifications and gene expression. Whereas traditional CGH uses DNA as test and reference in hybridisations, expressive genomic hybridisation (EGH) uses globally amplified mRNA as test and normal DNA as reference. EGH is a rapid and powerful tool for localising and studying global gene expression profiles and correlating them with loci of genetic amplifications using traditional CGH. EGH was used to correlate genetic amplifications detected by CGH with the expression profile of two independent cell lines-Colo320 and T47D. Although many amplifications resulted in overexpression, other amplifications were partially or completely silenced at the cytogenetic level. This technique will assist in the analysis of overexpressed genes within amplicons and could resolve a controversial issue in cancer cytogenetics; namely, the relation between genetic amplifications and overexpression.


Cell therapy of cerebral palsy.
The paper presents the results of a controlled study of cell therapy in 30 patients with severe forms of cerebral palsy. A cell suspension from immature nervous and hemopoietic tissues was injected into the subarachnoidal space of a recipient through a spinal puncture. Immune sensitization to donor antigens (detected by suppression of lymphocyte migration) was noted in few patients. In none patients laboratory and clinical signs of tissue-destructive autoimmune reactions were observed. One year after treatment activity of the major psychomotor functions in treated patients considerably surpassed the normal. No delayed complications of cell therapy were noted. These findings suggest that cell therapy is an effective, safe, and immunologically justified method of therapy for patients with cerebral palsy.


Further investigations on the maturation and degeneration of human ovarian follicles and their oocytes.
Human ovarian follicles showed activity of alkaline phosphatase, acid phosphatase (AcP) leucine aminopeptidase, and lactate dehydrogenase (LDH) in the theca interna. The granulosa of nonovulatory tertiary follicles showed moderate activity, whereas that of preovulatory Graafian follicles showed strong activity of LDH and AcP. The activity of these enzymes in the follicular fluid was measured. In nonovulatory tertiary follicles, activity of 3beta-ol-steroid dehydrogenase (3beta-OHSD) was found only in the theca interna; their fluid contained 290 ng/ml of progesterone and 502 ng/ml of 17beta-estradiol (average). Preovulatory Graafian follicles showed activity of 3beta-OHSD in the theca as well as in the granulosa. The progesterone concentration of the fluid was 7037 ng/ml and the 17beta-estradiol concentration was 2800 ng/ml (average). More oocytes could be aspirated from ovaries of younger women than from those of older women. In both age groups one out of three oocytes was degenerated. Oocytes with preovulatory changes were found only in follicles with preovulatory changes in their walls. Degenerated oocytes were found in some nonovulatory follicles as well as in some follicles with preovulatory changes in their walls.


Quality of life in patients undergoing ileal pouch-anal anastomosis at the University of Cincinnati.
In 1978, Drs. Fischer and Martin were among the first to preserve anorectal continence and create a pelvic reservoir in adult patients, in what has become the ileal pouch-anal anastomosis (IPAA). Here we review our institutions' experience with 379 of these procedures from 1978 to present. To assess the specific health concerns of patients with ulcerative colitis (UC) and familial adenomatous polyposis (FAP) and determine the effects of IPAA on health-related quality of life, we evaluated patients with the SF-36, the Rating Form of Inflammatory Bowel Disease Patient Concerns (RFIPC), time trade-off questions, and a gamble question. IPAA patients did not differ from the general population in seven of eight general health categories assessed by the SF-36. When compared with the UC population as a whole using the RFIPC they had reduced concerns in almost all areas. In addition, time trade-off and gamble questions indicated that these patients, as a group, are willing to accept a significant risk of dying in order to achieve their results of the IPAA. This high level of satisfaction has led to the referral of patients who would not have otherwise considered a procedure requiring permanent ileostomy.


Phase I trial of cisplatin and razoxane (ICRF-159) (NSC #119875) in advanced squamous cell carcinoma of the cervix. A Gynecologic Oncology Group Pilot Study.
Both cisplatin and razoxane have shown activity in squamous carcinoma of the cervix. Cisplatin at a dose of 50 mg/m2 intravenously every 21 days was combined with razoxane at two dose levels--750 mg/m2 weekly and 1150 mg/m2 weekly. Three patients were treated at the first dose level and six patients were treated at the second dose level of razoxane. No objective regressions were observed, and three patients refused to continue therapy at the higher dose of razoxane because of nausea and vomiting. Further study of this regimen in cervical cancer is not recommended.


Case files of the medical toxicology fellowship of the California poison control system-San Francisco: calcium plus digoxin-more taboo than toxic?
A 68 year old woman with a history of diabetes mellitus presented to the emergency department after experiencing several days of nausea and vomiting. Recorded vital signs included: blood pressure 98/32 mm Hg, heart rate 69 bpm, "normal" respirations, and no fever. Her initial ECG revealed peaked T waves and regular rhythm with no visible P waves. Initial serum electrolytes were notable for a potassium level of 7.7 mmol/L, and a creatinine level of 9.6 mg/dL (849 micromol/L). She was unable to provide a list of her current medications or other medical history. There was no old chart immediately available for comparison.


[Clinical characteristics and the surgical treatment of patients with perforated gastroduodenal ulcers].
The peculiarities of diagnosis and treatment of perforated gastric and duodenal ulcer in 367 patients with regard to age, history of the disease were studied. The expediency of early surgical treatment of the ulcer disease is noted.


Lorlatinib Tolerability and Association With Clinical Outcomes in Patients With Advanced ALK- or ROS1-Rearranged NSCLC: A Brief Report.
Treatment with lorlatinib for patients with advanced ALK- and ROS1-rearranged NSCLC (ALK+ and ROS1+ NSCLC) is associated with a unique set of adverse events (AEs) often requiring dose reduction. However, the impact of dose reductions on outcomes remains unclear and is mainly limited to analyses from prospective studies of lorlatinib in the first-line setting. We reviewed the course of 144 patients with advanced ALK- or ROS1-rearranged NSCLC treated with lorlatinib in the second-line or later setting to assess the frequency of dose reductions resulting from treatment-related AEs (TRAEs) and the association between dose reductions and progression-free survival (PFS) and overall survival (OS). A total of 58 patients (40%) had TRAE-related dose reductions, most (59%) owing to neurocognitive AEs or neuropathy. Among all patients, the median PFS was 8.1 months (95% confidence interval [CI]: 6.4-11.8); the median OS was 20.7 months (95% CI: 16.3-30.5). Among patients who were started on lorlatinib 100 mg/d (n = 122), a Cox regression model with the occurrence of a dose reduction as a time-dependent covariate indicated no association between dose reduction and PFS (hazard ratio = 0.86, 95% CI: 0.54-1.39) or OS (hazard ratio = 0.78, 95% CI: 0.47-1.30). Lorlatinib dose reductions were not associated with inferior clinical outcomes in this multicenter analysis. Prompt identification of lorlatinib TRAEs and implementation of dose reductions may help maximize tolerability without compromising outcomes.


Gene frequency and haplotype distribution of hemoglobin E among seven minority groups of Yunnan, China.
The aim of this study was to determine the distribution and origin of hemoglobin E (HbE) in seven minority groups from various geographical regions of the malaria-endemic Yunnan province, southwestern China, which have similar ethnic origins and geographic relationships with HbE-prevalent populations of Southeast Asian countries. By using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) methods, the prevalence of HbE was examined in 1488 individuals from seven native minority groups of Yunnan, and β-globin gene cluster haplotypes were determined on 1420 chromosomes. The prevalence of HbE in the study populations ranged from 1.5 to 39.1%. Higher HbE prevalence was correlated with the minority groups of Tibeto-Burman origin and groups from the Dehong district. The βE -globin genes in Yunnan were mostly associated with three haplotypes [-+++++-], [+----+-], and [-+-+++-] on chromosomes with gene framework 2. Interestingly, the predominant βE associated haplotype in Yunnan minorities was remarkably different from that in other previously reported populations. This study, for the first time, reports population-based data on the heterogeneity of HbE gene frequencies and haplotype distribution in native minorities from southwestern China. Natural selection based on the presence of malaria, ethnic origin, and epistatic interactions may be factors of varying importance for the remarkable variation in HbE frequency among these minority groups. In addition, there appears to be a common origin of the βE -globin gene in populations from Yunnan and Southeast Asia. Am. J. Hum. Biol. 28:927-931, 2016. © 2016Wiley Periodicals, Inc.


Thiazolo[4,5-d]pyrimidines as a privileged scaffold in drug discovery.
Thiazolo[4,5-d]pyrimidines are fused heterocyclic ring-systems that can be viewed at the first glance as purine isosteres. They are the 7 thia-analogs of purines via the replacement of the nitrogen at position 7 of the purine ring by a sulfur atom. Because of the structural resemblance to adenine and guanine and their related derivatives as adenosine, guanosine, cAMP, cGMP and similar biomolecules, many thiazolo[4,5-d]pyrimidines scaffold were developed and utilized by medicinal chemists to design novel therapeutics. Many were found to have a broad range of pharmacological activities. The outstanding development of thiazolo[4,5-d]pyrimidines within a short time span shows its magnitude of usefulness for medicinal chemistry research. Despite their importance from pharmacological and synthetic point of views, hardly there is a comprehensive review of thiazolo[4,5-d]pyrimidines applications in medicinal research to date. Thus, this review article describes the structures and medicinal significance of all classes of thiazolo[4,5-d]pyrimidines reported in literature to date. It describe the development of thiazolo[4,5-d]pyrimidines as immune-modulators, Corticotropin Releasing Factor (CRF) receptor antagonists, anti-Parkinson's, antiviral, anticancer, antibacterial, antifungal, analgesic, anti-inflammatory agents including COX inhibitors, chemokines antagonists and Fractlkine receptor antagonists.