Pulmonary arterial hypertension in the southern hemisphere: results from a registry of incident Brazilian cases.
Pulmonary arterial hypertension (PAH) is a rare and ultimately fatal disorder of the pulmonary vasculature. There is increasing interest in the worldwide characteristics of patients with PAH, although data coming from the Southern Hemisphere remain scarce. The objective of this study was to describe a cohort of incident patients with PAH from a large reference center in Brazil. All consecutive patients who received a diagnosis of PAH by right-sided heart catheterization between 2008 and 2013 were included in the study. A total of 178 patients with newly diagnosed PAH were enrolled in the study (mean age, 46 years; female/male ratio, 3.3:1; 45.5% in New York Heart Association functional class III or IV). Idiopathic PAH (IPAH), connective tissue disease (CTD), and schistosomiasis-associated PAH (Sch-PAH) accounted for 28.7%, 25.8%, and 19.7% of all cases, respectively. The patients were treated with phosphodiesterase type 5 inhibitors (66%), endothelin receptor antagonists (27%), or a combination of both (5%). For the PAH group as a whole, the estimated survival rate 3 years after diagnosis was 73.9%. The prognosis for the patients with CTD was worse than that for the patients with IPAH and Sch-PAH (P = .03). The distribution of PAH causes and the baseline characteristics in our registry clearly differ from the previously published European and US-based registries. These differences highlight the importance of regional registries and also raise questions regarding the need to better account for such differences in future clinical trials.


Multiple genetic variant association testing by collapsing and kernel methods with pedigree or population structured data.
Searching for rare genetic variants associated with complex diseases can be facilitated by enriching for diseased carriers of rare variants by sampling cases from pedigrees enriched for disease, possibly with related or unrelated controls. This strategy, however, complicates analyses because of shared genetic ancestry, as well as linkage disequilibrium among genetic markers. To overcome these problems, we developed broad classes of "burden" statistics and kernel statistics, extending commonly used methods for unrelated case-control data to allow for known pedigree relationships, for autosomes and the X chromosome. Furthermore, by replacing pedigree-based genetic correlation matrices with estimates of genetic relationships based on large-scale genomic data, our methods can be used to account for population-structured data. By simulations, we show that the type I error rates of our developed methods are near the asymptotic nominal levels, allowing rapid computation of P-values. Our simulations also show that a linear weighted kernel statistic is generally more powerful than a weighted "burden" statistic. Because the proposed statistics are rapid to compute, they can be readily used for large-scale screening of the association of genomic sequence data with disease status.


Chrysophyllum cainito stem bark extract induces apoptosis in Human hepatocarcinoma HepG2 cells through ROS-mediated mitochondrial pathway.
Hepatocellular carcinoma is the most common type of primary liver cancer in humans. This study aimed to demonstrate anticancer properties of an aqueous extract from Chrysophyllum cainito stem bark (CE) and its underlying mechanisms. Our MTT assay results showed that CE significantly reduced human hepatocellular carcinoma (HepG2) cell viability with the IC50of 100 µg/mL, while human dermal primary fibroblast (HDFa) cells showed less susceptibility in every concentration tested. Determined by Annexin V staining, the proportion of apoptotic HepG2 cells increased in a dose-dependent fashion after 24 hour-exposure of CE. The results from Western blot analysis confirmed that CE reduced procaspase-3, suggesting apoptosis by activating caspase-3 cleavage. Using the DCFH-DA and DiOC6 fluorescent probes, it was found that CE significantly stimulated the generation of reactive oxygen species (ROS) and reduced mitochondrial membrane potential (Δψ m), respectively. According to cell cycle analysis, CE (100 µg/mL) profoundly increased the percentage of cells in the sub-G1 phase, indicating cell apoptosis. These data suggest that CE induces apoptosis and cell death in human hepatocellular carcinoma via generation of intracellular ROS and disruption of Δψm. This is the first demonstration of the anticancer activity with proposed underlying mechanism of CE in liver cancer cells.


Asymptotic properties of the inhomogeneous Lotka-Von Foerster system.
"In this paper, we investigate an extension of the multistate stable population model, which makes allowances for migration. The model is formulated as an inhomogeneous system of first order partial differential equations with integral boundary conditions. First, we construct its classical solution. Next, we reformulate the system as an abstract inhomogeneous Cauchy problem on a Banach space, and give its mild solution by using the population semigroup. Our main purpose is to investigate the asymptotic behavior of the mild solution." (SUMMARY IN FRE)


The complete mitochondrial genome of Sillago aeolus (Perciformes: Sillaginidae).
The long PCR and primer walking methods are used for determining the complete mitochondrial genome sequence of Sillago aeolus. The entire sequence is 16,499 bp in length including 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and 2 non-coding regions (L-strand replication origin and control region). Within the control region, we identified the termination-associated sequence domain (TAS), central conserved domains (CSB-F, CSB-E, CSB-D, CSB-C, CSB-B and CSB-A), and three conserved sequence blocks (CSB-1, CSB-2 and CSB-3).


Biological activity of two new pyrrole derivatives against stored-product species: influence of temperature and relative humidity.
Members of the pyrrole group are likely to have interesting properties that merit additional investigation as insecticides at the post-harvest stages of agricultural commodities. In the present work, the insecticidal effect of two new pyrrole derivatives, ethyl 3-(benzylthio)-4,6-dioxo-5-phenyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-carboxylate (3i) and isopropyl 3-(benzylthio)-4,6-dioxo-5-phenyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-carboxylate (3k) were studied as stored-wheat protectants against two major stored-product insect species, the confused flour beetle, Tribolium confusum Jaquelin du Val adults and larvae and the Mediterranean flour moth, Ephestia kuehniella Zeller larvae at different doses (0.1, 1 and 10 ppm), exposure intervals (7, 14 and 21 days), temperatures (20, 25 and 30°C) and relative humidity (55 and 75%) levels. For T. confusum adults, in the case of the pyrrole derivative 3i, mortality was low and it did not exceed 32.2% in wheat treated with 10 ppm 3i at 30°C and 55% relative humidity. Progeny production was very low (<1 individual/vial) in all combinations of 55% relative humidity, including control. In the case of the pyrrole derivative 3k, mortality reached 67.8% at 30°C and 55% relative humidity in wheat treated with 10 ppm after 21 days of exposure. Progeny production was low in all tested combinations (≤0.7 individuals/vial) of 55% relative humidity, including control. For T. confusum larvae, in the case of the pyrrole derivative 3i, at the highest dose, mortality was 82.2% at 25°C and 55% relative humidity whereas in the case of 3k it reached 77.8% at the same combination. In contrast, mortality at 75% relative humidity remained very low and did not exceed 13.3%. For E. kuehniella larvae, the highest mortalities, 44.4 and 63.3%, were observed in 10 ppm at 25°C and 55% relative humidity for both pyrrole derivatives. The compounds tested here have a certain insecticidal effect, but this effect is moderated by the exposure, the target species, the temperature and the relative humidity.


Is there a role for 'modified VAD' in the treatment of multiple myeloma?
VAD, (Vincristine, Doxorubicin and Dexamethasone) was initially proposed as a salvage therapy for myeloma patients in whom prior alkylating agent therapy failed, although in recent years VAD has been surpassed by novel combination therapies with new biological agents such as thalidomide (and its derivative, lenalidomide) and bortezomib. After the excellent results obtained by the novel agents, VAD can no longer be proposed in preparation to autologous transplantation, although there are still indications that VAD remains useful and clinically relevant in the initial treatment of symptomatic multiple myeloma.


Endoluminal grafting for aneurysmal and occlusive disease in the superficial femoral artery: early experience.
To examine whether endoluminal grafts (ELGs) of radically expandable polytetrafluoroethylene (PTFE) can successfully form durable internal conduits to revascularize lengthy occlusive disease and exclude aneurysms in the femoropopliteal (FP) arteries. Under protocol, implantation of an unpredilated PTFE tube ELG anchored with Palmaz stents using a low-profile percutaneous delivery system was attempted in 50 symptomatic patients for a variety of pathologies: (1) restenosis; (2) complex lesions unlikely to be treated successfully with other endoluminal therapies; (3) acute angioplasty failure; and (4) aneurysms. There were 37 occlusions, 14 stenoses, and 2 long, combined stenotic-aneurysmal lesions in 47 native arteries, 5 FP grafts, and 1 femorotibial (FT) vein graft. Thirty-two percent of the patients had < or = vessel runoff. The average lesion length was 20.4 +/- 11.4 cm (range 1.5 to 40), and the mean preoperative ankle-brachial index (ABI) at rest was 0.53 +/- 0.14. In a 20-month period through April 1995, 50 patients (34 males, 16 females; mean age 69.5 years, range 45 to 87) underwent 54 procedures in 53 limbs; 55 ELGs were successfully deployed in 51 limbs; 2 patients were converted to FP bypass owing to technical problems (96% procedural success). There were 18 inhospital complications: 1 distal wire dissection repaired with an additional ELG; 2 hematomas requiring surgical repair; 1 graft collapse; 1 pseudoaneurysm at the site of a mid-ELG leak; 7 minor access sequelae; and 6 acute ELG thromboses, 4 treated with lytic therapy and balloon dilation, 1 with open thrombectomy, and 1 with bypass grafting. The mean postoperative ABI was 1.01 +/- 0.10. During the 30-day postprocedure period, 2 ELGs rethrombosed and 2 other limbs (3 ELGs) occluded; 1 thrombosis and 1 rethrombosis were lysed successfully, but the other 2 patients had an FP bypass. Over the mean 8.3 +/- 5.5 month follow-up, 6 additional ELGs occluded and 1 reoccluded; 4 were successfully treated by endovascular techniques for a primary patency of 72% and secondary patency of 84% by life-table analysis. Endovascular grafting is a conceptually attractive technique that has the potential to expand the current boundaries of interventional treatment. This preliminary experience attests to the feasibility and safety of ELG deployment in the superficial femoral arteries. Whether such a device can match the durability of classical revascularization techniques remains to be determined in clinical trials when device configurations and deployment techniques have been standardized.


Commentary: Emerging technologies oversight: research, regulation, and commercialization.
This paper reviews the paper by Kuzma, Najmaie, and Larson that looks at what can be learned from the experience with genetically engineered organisms for oversight of emerging technologies more generally. That paper identifies key attributes of a good oversight system: promoting innovation, ensuring safety, identifying benefits, assessing costs, and doing so all while building public confidence. In commenting on that analysis, this paper suggests that looking at "oversight" in three phases - research and development, regulatory review, and market acceptance - can help to determine when certain of these attributes should take precedence over others and how to structure remedies when an error occurs. The result is an approach that is precautionary with respect to research and development, prudent and open to public input in the regulatory review stage, and purposefully persuasive once market acceptability is at stake, with remedies that are risk-containing in the first phase, risk-managing in the second, and risk-assuaging in the third. Combining the key attributes with the idea of three phases can help attune oversight to society's needs.


Preparation and characterisation of graphene oxide containing block copolymer worm gels.
This paper reports a generic method for preparing reinforced nanocomposite worm-gels. Aqueous poly(glycerol monomethacrylate)-b-poly(2-hydroxypropyl methacrylate) (PGMA-PHPMA) and methanolic poly(glycerol monomethacrylate)-b-poly(benzyl methacrylate) (PGMA-PBzMA) worm gels were prepared by RAFT-mediated polymerisation-induced self-assembly (PISA). The former system undergoes a reversible worm-to-sphere degelation transition upon cooling to 5 °C whilst the latter system undergoes the same transition on heating to 56 °C. This transition allows these copolymer dispersions to be readily mixed with graphene oxide (GO) whilst in a low viscosity state and form nanocomposite gels on returning to room temperature via a sphere-to-worm transition. Various quantities of GO were added to the studied copolymer dispersions at a fixed copolymer content of 15% w/w. A general trend was observed whereby relatively small quantities of GO caused the gel strength of the nanocomposite gel to be higher than that of the pristine worm-gel, as determined by oscillatory rheology. Additional quantities of GO resulted in gel weakening or prevented gel-reformation altogether. For instance, 15% w/w PGMA52-PHPMA130 worm gels had a storage modulus (G') of approximately 1.5 kPa. The addition of 1.5% w/w GO based on the copolymer caused G' to increase to approximately 4.0 kPa but >1.5% w/w GO resulted in gel strengths <1.0 kPa. A combination of aqueous electrophoresis and transmission electron microscopy measurements were used to investigate the mechanism of nanocomposite gel formation. It was observed that the PGMA-based copolymers readily absorb onto the surface of GO. Thus, the role of GO is both to strengthen the worm-gels when an optimal concentration of GO is used, but also prevent worm-reformation if too much copolymer becomes absorbed on the surface of the sheets.


Estimating exposed pulp lengths of tusks in the African elephant (Loxodonta africana africana).
Captive and wild African elephants frequently suffer tusk fractures. Several institutions shorten the tusks of captive elephants to reduce fractures and injury as a result of behaviour within enclosures. Fracturing or coronal amputations that expose pulp lead to pain for the elephant. Estimating coronal pulp lengths may thus help to minimise the risk of pulp exposure during amputations. We aimed to determine the length of the pulp beyond the lip margin from an external tusk characteristic. Tusks collected from elephants in Namibia and the Kruger National Park had similar morphological relationships. This statistical property allowed us to correct for missing data in our data sets. Pulp volume and pulp length correlated with tusk circumference at the lip. Even so, the circumference at the lip could not predict the length of the pulp in the crown external to the lip. Our findings suggest that tusks, irrespective of sex or age, amputated further than 300 mm from the lip should not expose pulp.


Halogen versus high-intensity light-curing of uncoated and pre-coated brackets: a shear bond strength study.
To evaluate the shear bond strengths of adhesive pre-coated brackets (APC) and conventional uncoated brackets (Victory) cured with two different light-curing units: a conventional halogen light (Visilux 2) and a micro-xenon light (Aurys). Ex vivo study Sixty freshly extracted bovine permanent mandibular incisors were randomly assigned to one of four groups, each group consisting of 15 specimens. Two groups (one for each type of bracket) were exposed to the halogen light for 20 seconds and used as controls. The remaining two groups were cured with the micro-xenon light for 2 seconds. After 24 hours, all samples were tested in a shear mode on an Instron Machine. Analysis was by two-way ANOVA with Scheffé's test for comparisons, Kaplan-Meier survival estimates, and Cox model. The Chi-square (chi(2)) test was used to determine significant differences in the ARI scores. The mean shear bond strength of the uncoated brackets cured with Visilux 2 was significantly higher than those of all the other groups tested. Both groups cured with Visilux 2 produced significantly higher mean shear bond strengths than those of the corresponding groups cured with Aurys. No statistically significant differences were found between the two groups cured with Aurys. Compared to halogen light-curing, the micro-xenon light enables the clinician to reduce significantly the curing time of both APC and uncoated brackets, and although significantly lower, their shear bond strengths may be clinically acceptable.


Lesions of the basolateral amygdala disrupt conditioning based on the retrieved representations of motivationally significant events.
One recent perspective (Blundell et al., 2001; 2003; Killcross and Blundell, 2002; Balleine et al. 2003) on the function of the basolateral region of the amygdala (BLA) suggests that it plays an important role in the representation of the sensory features of motivationally significant events. This predicts that lesions of the BLA will not produce a decrement in performance in conditioning procedures based on the formation of associations between the sensory aspects of neutral events but will interfere with conditioning based on associations between neutral cues and motivationally significant events. This prediction is supported by the evidence that BLA lesions were without effect on a sensory preconditioning procedure (experiment 1A) that used neutral cues but that BLA lesions did significantly impair representation-mediated conditioning (experiment 1B) when the target cues were motivationally significant at the time of training. These results demonstrate that animals with lesions of the BLA can represent the sensory aspects of neutral events but not the sensory aspects of motivationally significant events.


Phylogeography of two European Reticulitermes (Isoptera) species: the Iberian refugium.
Chemical, i.e. cuticular hydrocarbons, and molecular data were used to probe the phylogeography of Reticulitermes termites collected from various parts of France, Spain and Portugal. Phylogenetic relationships were inferred from sequences of the internal transcribed spacer (ITS2) of nuclear ribosomal RNA genes as well as from two partial mitochondrial DNA segments, the cytochrome oxidase II gene and a sequence combining the tRNA-Leu gene and fragments of the NADH dehydrogenase I and ribosomal 16S genes. Two species, namely, R. grassei and R. banyulensis, were identified based on an analysis of cuticular hydrocarbons and the identification was confirmed by ITS2 haplotyping. However, phylogeny based on the analysis of mitochondrial DNA was not completely in agreement with the conclusions drawn from the chemical and nuclear data. An analysis of 56 R. grassei colonies revealed intraspecific differentiation into two major lineages with distinct geographical ranges. Whereas analysis of cuticular hydrocarbons showed that R. banyulensis was chemically distinct from R. grassei, analysis of mitochondrial DNA showed its close kinship with the R. grassei lineage occurring in southern Spain. This kinship could be explained by their evolution from a common polymorphic ancestor species in this ice age refugium.


A CT-guided stereotactic system with direct verification of the centrage of the target before introducing the needle into the brain.
The author reports his experience with a stereotactic apparatus which gives more protection to patients during stereotactic operations. This apparatus allows the verification of the centrage of the target before puncturing the brain with the needle. In the most common stereotactic systems there is a headframe with three or four pins for the fixation to the skull. This apparatus is more simple: there is no frame and there is only one point of fixation to the skull: a small, L-shaped, aluminum bar, fixed in the right prefrontal region with two small screws. Because this system has only one point of fixation to the skull, it is movable like a bell. Besides the possibility of a direct verification of the centrage of the target, that has induced the author to study a new stereotactic apparatus for CT-guided stereotaxy, the small size of the apparatus means that it does not interfere with surgery in microneurosurgical stereotactic directed operations. This stereotactic apparatus can be used with CT-scan because it is aluminium with some parts in brass. The system, moreover, can be used whether scanner-dependent or scanner-independent, using two different softwares for calculation of the centrage of the target.


Acute gastroenteritis in a pediatric hospital in rio de janeiro in pre- and post-rotavirus vaccination settings.
A 4(1/2)-year hospital-based survey was conducted in Rio de Janeiro to determine baseline rates of gastroenteritis-related cases, hospitalizations, and deaths; to examine the prevalence of rotavirus strains causing admissions; and to assess the immediate impact of the nationwide rotavirus immunization program launched in March 2006. From August 2002 to May 2007, 14,473 (10.4%) of the 139,747 consultations had AGE as primary diagnosis, 491 (3.4%) children required hospitalization and two (0.4%) dehydrated children died. Gastroenteritis contribution to hospitalizations varied from ~2.3% in 2004 and 2006 to 6.4% in 2005, being roughly half of them rotavirus-related cases. A gradual decrease in rotavirus strain diversity was observed from 2002 to 2005 when a single G9P[8] prevailed until April 2006. Then only short profile G9P[4] and G2P[4] strains were detected. Gastroenteritis cases were distributed year-round in a trimodal pattern with major winter peaks. Local climate apparently affected the incidence of gastroenteritis: reduction in dry years (2004 and 2007) and explosive outbreaks caused by multiple agents during the heavy rainfalls and recurrent floods of the 2005-early 2006 period. Besides rotavirus, adenovirus and calicivirus were major gastroenteritis agents of these seemingly waterborne outbreaks. In conclusion, rotavirus vaccination impacted marginally, if at all, on the incidence of childhood gastroenteritis, as favorable results obtained by comparing data from the post-vaccine period to the preceding unusual 2005 year all but disappeared when comparing to previous pre-vaccination periods, and the shift towards G2P[4] rotavirus strains may be a global trend unrelated to vaccination.


Myocardial ischemia and infarction: growth of ideas.
This report reviews the author's involvement in the growth of ideas and basic concepts in myocardial ischemia resulting in the histological changes of myocardial infarction. Concepts arising from the study of myocardial substrate utilization, activation of the inducible form of nitric oxide synthase and production of prostacyclin and thromboxane in the infarcted heart are presented. New approaches are discussed dealing with the effects of nonsteroidal anti-inflammatory drugs on myocardial production of nitric oxide and prostanoids, and with the relevance of the inducible form of cyclooxygenase. The review also records a number of significant similarities between angiogenesis in the ischemic heart and some cancers. Angiogenesis in both instances originates from inflammatory reactions, illustrating how different tissues and organs such as ischemic heart muscle and cancer react to similar pathological stimuli in an identical manner. This multifocal approach opens new concepts on myocardial ischemia and cancer.


In vivo bioluminescent monitoring of chemical toxicity using heme oxygenase-luciferase transgenic mice.
Transgenic mice expressing the luciferase (luc) gene under the control of the heme oxygenase-1 promoter (Ho1) were used to measure the induction of heme oxygenase in response to known toxicants. Transgenic Ho1-luc expression was visualized in vivo using a low-light imaging system (IVIS). Ho1-luc activation was compared to Ho1-luc expression, HO1 protein levels, standard markers of toxicity, and histology. Male and female Ho1-luc transgenic mice were exposed to acute doses of cadmium chloride (CdCl2, 3.7 mg/kg), doxorubicin (15 mg/kg), and thioacetamide (300 mg/kg). These agents induced the expression of Ho1-luc in the liver and other tissues to varying degrees. The greatest increase in Ho1-luc activity was observed in the liver in response to CdCl2; intermediate responses were observed for doxorubicin and thioacetamide. Induction of the Ho1-luc transgene by these agents was similar to endogenous protein levels of heme oxygenase as assessed by Western blotting, and generally correlated with plasma levels of circulating enzymes reflecting hepatic or general tissue damage. Histopathology confirmed the toxic effects of CdCl2 on liver and kidney; doxorubicin on kidney, liver, and intestine; and thioacetamide on the liver. Tissue damage was much more pronounced than the luciferase expression following thioacetamide treatment when compared with tissue damage and bioluminescence of the other toxicants. Nevertheless, the induction of Ho1-luc expression following exposure to these agents suggests that the Ho1-luc transgenic mouse may prove useful as a model for in vivo screening of compounds that induce luciferase expression as a marker of toxicity.


[Pediatric liver transplantation--experience at Schneider Children's Medical Center].
Introduction of segmental graft transplantation from living donors and split livers from cadaver donors has led to major advances in liver transplantation (LTx) in children. To report our initial experience with pediatric LTx performed at our center. Data collection on all children undergoing LTx between the years 1996-2003 including the analysis of the graft and patient survivals and reports of complications. Forty LTx were performed in 38 children at the mean age of 6.2 years, including two retransplants. There were 15 whole liver allografts and 25 segmental grafts including: 12 living donor grafts, 5 splits and 8 reduced grafts from cadaver donors. At 40 months mean follow-up period, patient and graft survival were 81% and 72.5%, respectively. There was post-transplant mortality in seven cases--5 children died during the first month and two children passed away after 6 months (recurrent disease) and 14 months (metastatic tumor). Vascular complications included: one early and one late portal vein thrombosis (5%) and six cases of hepatic artery thrombosis (15%). In the latest group, 3 grafts were salvaged by thrombectomy and another 3 children underwent re-transplantation. There were two bile leaks (5%) and 6 bile duct strictures (15%). The bile-duct strictures were successfully corrected by surgery in one child and by transhepatic dilatation in another 4 children. One child remained with intrahepatic strictures in one of the two hepatic segments. The use of segmental liver allografts enables the performance of pediatric liver transplantation in Israel. Gathered experience and enhanced skills will ensure improved results over time.


Specific detection of Trichophyton rubrum and Trichophyton interdigitale based on loop-mediated isothermal amplification (LAMP) from onychomycosis specimens.
In diagnosing onychomycosis, diseases with similar features must be excluded by demonstrating the presence of fungal infection and identifying the fungal species. However, fungal culture of onychomycosis-derived samples usually takes many weeks to yield species identification results, and is associated with a low successful culture rate. Loop-mediated isothermal amplification (LAMP) is a highly sensitive and specific molecular biological method that can amplify DNA at a constant temperature, allowing for a simpler testing procedure, shorter detection time and less cost than conventional techniques including quantitative polymerase chain reaction. We have developed a new LAMP method specifically to detect Trichophyton rubrum (T. rubrum) and Trichophyton interdigitale (T. interdigitale), major causative dermatophytes for onychomycosis, and analyzed the correlation between LAMP results and those of the existing fungal culture method for the detection and identification of Trichophyton species from onychomycosis-derived samples. The results showed that all 59 specimens in which T. rubrum or T. interdigitale was identified by fungal culture also tested positive by LAMP, giving a 100% positivity concordance rate between the two methods. Moreover, all 55 and four specimens in which T. rubrum and T. interdigitale were identified by fungal culture, respectively, also tested positive for each species by LAMP, again giving a 100% species-identification concordance rate. The high correlation demonstrated between LAMP and fungal culture results in detection and identification of Trichophyton species from onychomycosis-derived samples suggests high reliability of LAMP as a promising, alternative mycological detection and identification technique which can serve as an alternative to the fungal culture method.


Animal models got you puzzled?: think pig.
Swine are an excellent large animal model for human health and disease because their size and physiology are similar to humans, in particular, with respect to the skin, heart, gastrointestinal tract, and kidneys. In addition, the pig has many emerging technologies that will only enhance the development of the pig as the nonrodent biomedical model of choice.


Platelet-Neutrophil Interactions Are Lower in Cord Blood of Premature Newborns.
To quantify platelet-neutrophil interaction by flow cytometry, in newborn cord blood, as a function of gestational age. Little is known about platelet function markers in the newborn, and developmental variations in these markers are not well described. Cord blood samples were obtained from 64 newborns between 23 and 40 weeks' gestation. The neonates were grouped into three categories: preterm (< 34 weeks' gestation, n = 21), late preterm (34 to < 37 weeks' gestation, n = 22), and term (≥37 weeks' gestation, n = 21). We monitored the expression of P-selectin and the formation of platelet-neutrophil aggregates (PNAs) by flow cytometry while using adenosine 5'-diphosphate (ADP) or thrombin receptor-activating peptide (TRAP) as agonists. PNAs were significantly lower in preterm compared to term neonates after TRAP or ADP stimulations (11.5 ± 5.2% vs. 19.9 ± 9.1%, p < 0.001, or 24.0 ± 10.1% vs. 39.1 ± 18.2%, p = 0.008, respectively). The expression of P-selectin also tended to be lower in preterm neonates, with significant positive correlations between P-selectin expression and PNA formation. The potential formation of PNAs correlates with gestational age. This suggests that the development of functional competencies of platelets and neutrophils continues throughout gestation, progressively enabling interactions between them.


The fatty acid composition of colostrum in three geographic regions of China.
To describe the fatty acid composition of human colostrum in three different regions of China. Women were recruited from river/lake, coastal and inland regions of China during late pregnancy. Their diet frequency was assessed during the 34th week of pregnancy. Colostrum samples were collected between postpartum days 3 and 5. Thirty-one different fatty acids were separated and identified in colostrum. There were significant differences among the women in the three regions with respect to food and culinary oil consumption patterns. The mean±SD fat content (g/100 g) of colostrum from river/lake, coastal and inland regions was 2.17±1.26, 2.50±1.39 and 2.68±1.26, respectively (p=0.265, adjusted by pregnant women's age). The main saturated, monounsaturated, n-6 polyunsaturated and n-3 polyunsaturated fatty acids in colostrum were 16:0, 18:1n9, 18:2n6 and 18:3n3, respectively. Colostrum from river/lake region had the lowest composition of total SFA and of total n-6 PUFA and the highest composition of total MUFA and of total n-3 PUFA. The amount of DHA (22:6n3) in colostrum (mean±SD, % wt/wt) was 0.51±0.18, 0.52±0.20 and 0.35±0.13 in the river/lake, coastal and inland regions, respectively (p=0.0002, adjusted by pregnant women's age). Frequency of maternal marine fish intake during the late pregnancy was positively correlated with DHA in colostrum (r=0.203, p=0.040). The fatty acid composition of colostrum differs across geographical regions in China, which may be related to differences in maternal dietary pattern.


FITC-conjugated cyclic RGD peptides as fluorescent probes for staining integrin αvβ3/αvβ5 in tumor tissues.
This study sought to evaluate FITC-conjugated cyclic RGD peptides (FITC-RGD2, FITC-3P-RGD2, and FITC-Galacto-RGD2) as fluorescent probes for in vitro assays of integrin αvβ3/αvβ5 expression in tumor tissues. FITC-RGD2, FITC-3P-RGD2, and FITC-Galacto-RGD2 were prepared, and their integrin αvβ3/αvβ5 binding affinity was determined using the displacement assay against (125)I-echistatin bound to U87MG glioma cells. IC50 values of FITC-Galacto-RGD2, FITC-3P-RGD2, and FITC-RGD2 were calculated to be 28 ± 8, 32 ± 7, and 89 ± 17 nM, respectively. The integrin αvβ3/αvβ5 binding affinity followed a general trend: FITC-Galacto-RGD2 ∼ FITC-3P-RGD2 > FITC-RGD2. The xenografted tumor-bearing models were established by subcutaneous injection of 5 × 10(6) tumor cells into shoulder flank (U87MG, A549, HT29, and PC-3) or mammary fat pad (MDA-MB-435) of each athymic nude mouse. Three to six weeks after inoculation, the tumor size was 0.1-0.3 g. Tumors were harvested for integrin αvβ3/αvβ5 staining, as well as hematoxylin and eosin (H&E) staining. Six human carcinoma tissues (colon cancer, pancreatic cancer, lung adenocarcinoma, squamous cell lung cancer, gastric cancer, and esophageal cancer) were obtained from recently diagnosed cancer patients. Human carcinoma slides were deparaffinized in xylene, rehydrated with ethanol, and then used for integrin αvβ3/αvβ5 staining, as well as H&E staining. It was found that the tumor staining procedures with FITC-conjugated cyclic RGD peptides were much simpler than those with the fluorescence-labeled integrin αvβ3 antibodies. Since FITC-RGD2, FITC-3P-RGD2, and FITC-Galacto-RGD2 were able to co-localize with the fluorescence-labeled integrin β3 antibody, their tumor localization and tumor cell binding are integrin αvβ3-specific. Quantification of the fluorescent intensity in five xenografted tumors (U87MG, MDA-MB-435, A549, HT29, and PC-3) and six human carcinoma tissues revealed an excellent linear relationship between the relative integrin αvβ3/αvβ5 expression levels determined with FITC-Galacto-RGD2 and those obtained with the fluorescence-labeled anti-human integrin β3 antibody. There was also an excellent linear relationship between the tumor uptake (%ID/g) of (99m)Tc-3P-RGD2 (an integrin αvβ3/αvβ5-targeted radiotracer) and the relative integrin αvβ3/αvβ5 expression levels from the quantification of fluorescent intensity in the tumor tissues stained with FITC-Galacto-RGD2. These results suggest that FITC-conjugated cyclic RGD peptides might be useful to correlate the in vitro findings with the in vivo imaging data from an integrin αvβ3/αvβ5-targeted radiotracer. The results from this study clearly showed that the FITC-conjugated cyclic RGD peptides (particularly FITC-3P-RGD2 and FITC-Galacto-RGD2) are useful fluorescent probes for assaying relative integrin αvβ3/αvβ5 expression levels in tumor tissues.


The S locus glycoprotein and the S receptor kinase are sufficient for self-pollen rejection in Brassica.
Self-incompatibility (SI) is one of several mechanisms that have evolved to prevent inbreeding in plants. SI in Brassica is controlled by the polymorphic S locus complex. Two S locus-encoded proteins are coordinately expressed in the stigma epidermis: the cell wall-localized S locus glycoprotein (SLG) and the plasma membrane-anchored S receptor kinase (SRK). These proteins are thought to recognize a pollen factor that leads to the rejection of self-pollen. Evidence has accumulated that indicates that both proteins are necessary for the ability of the stigma to inhibit self-pollen. However, it has not been possible to prove this necessity definitively or to demonstrate that these genes are sufficient for this phenotype, because previous attempts to transfer this phenotype via transformation have not been successful. In this study, two overlapping S locus genomic clones, which cover approximately 55 kilobases of DNA and contain the SLG, SRK, and an anther-expressed gene in the region common to the two, were introduced into a self-compatible Brassica napus line. The resulting transgenic plants were shown to carry the female part of the SI phenotype, rejecting pollen in a haplotype-specific manner. However, the pollen SI phenotype was not found in any of the transgenic plants. These data show that the SLG and SRK are sufficient for the female side but not the male side of the SI phenotype in Brassica and that there must be an independent pollen S factor encoded outside the cloned region.


Structural Fluctuations of Aromatic Residues in an Apo-Form Reveal Cryptic Binding Sites: Implications for Fragment-Based Drug Design.
Cryptic sites are binding pockets that are transiently formed in an apo form or that are induced by ligand binding. The investigation of cryptic sites is crucial for drug discovery, since these sites are ubiquitous in disease-related human proteins, and targeting them expands the number of drug targets greatly. However, although many computational studies have attempted to identify cryptic sites, the detection remains challenging. Here, we aimed to characterize and detect cryptic sites in terms of structural fluctuations in an apo form, investigating proteins each of which possesses a cryptic site. From their X-ray structures, we saw that aromatic residues tended to be found in cryptic sites. To examine structural fluctuations of the apo forms, we performed molecular dynamics (MD) simulations, producing probability distributions of the solvent-accessible surface area per aromatic residue. To detect aromatic residues in cryptic sites, we have proposed a "cryptic-site index" based on the distribution, demonstrating the performance via several measures, such as recall and specificity. Besides, we found that high-ranking aromatic residues were likely to probe concaves in a cryptic site. This implies that such fluctuations provide a profile of scaffolds of compounds with the potential to bind to a particular cryptic site.


Blockade of maitotoxin-induced endothelial cell lysis by glycine and L-alanine.
The maitotoxin (MTX)-induced cell death cascade in bovine aortic endothelial cells (BAECs) is a model for oncotic/necrotic cell death. The cascade is initiated by an increase in cytosolic free Ca(2+) concentration ([Ca(2+)](i)), which is followed by the biphasic uptake of vital dyes. The initial phase of dye entry reflects activation of large pores and correlates with surface membrane bleb formation; the second phase reflects cell lysis. In the present study, the effect of the cytoprotective amino acid glycine was examined. Glycine had no effect on MTX-induced change in [Ca(2+)](i) or on the first phase of vital dye uptake but produced a concentration-dependent (EC(50) approximately 1 mM) inhibition of the second phase of dye uptake. No cytoprotective effect was observed with l-valine, l-proline, or d-alanine, whereas l-alanine was equieffective to glycine. Furthermore, glycine had no effect on MTX-induced bleb formation. To test the hypothesis that glycine specifically blocks formation of a lytic "pore," the loss of fluorescence from BAECs transiently expressing GFP and concatemers of GFP ranging in size from 27 to 162 kDa was examined using time-lapse videomicroscopy. MTX-induced loss of GFP was rapid, correlated with the second phase of dye uptake, and was relatively independent of molecular size. The MTX-induced loss of GFP from BAECs was completely blocked by glycine. The data suggest that the second "lytic" phase of MTX-induced endothelial cell death reflects formation of a novel permeability pathway that allows macromolecules such as GFP or LDH to escape, yet can be prevented by the cytoprotective agents glycine and l-alanine.


Relating self-disorders to neurocognitive and psychopathological measures in first-episode schizophrenia.
The notion of a disturbed self as the core feature of schizophrenia dates back to the founding texts on the illness. Since the development of the psychometric tool for examination of anomalous self-experience (EASE), self-disorders have become accessible to empirical research. Empirical studies have shown that EASE measured self-disorders predict schizophrenia spectrum in prospective studies and consistently show a selective hyper aggregation of self-disorder in schizophrenia and schizotypal disorders. The aim of this study is to investigate the relationship between self-disorders cognitive deficits and symptoms in schizophrenia. Thirty-five non-acute first-episode patients with schizophrenia and 35 matched healthy controls were evaluated with EASE, cognitive deficits, and symptoms (PANSS positive, negative and general). [Correction added on 28 January 2022, after first online publication: the words, 'evaluated with' were missing and have now been added to the preceding sentence.] RESULTS: The results show that self-disorders and symptoms are correlated among patients with schizophrenia, but not with cognitive deficits. Moreover, with the exception of attentional deficits, neurocognitive impairment was not significantly higher among patients with schizophrenia compared to healthy controls. We argue that this adds support to a view of schizophrenia as being characterized by specific traits of pre-reflective self-disturbance, which are related to the severity of symptoms, whereas neurocognitive impairment reflects a separate or distinct aspect of schizophrenia.


The Prevalence and Etiology of Surgical Site Infections Following Gastrointestinal Tract Surgery: A Cross-Sectional Study From a Tertiary Care Hospital.
Background Surgical site infection (SSI) is the most commonly occurring infection in postoperative patients. This study is conducted to evaluate the prevalence of SSI in patients following gastrointestinal tract surgery and identify the risk factors. Method A cross-sectional study was conducted at the Jinnah Postgraduate Medical Centre (JPMC), Karachi, Pakistan, between December 2021 and May 2022. A total of 132 patients participated in the study who were at least 18 years older and had undergone the gastrointestinal surgical procedure. Patients who refused to give consent, died after the procedure, and were diagnosed with SSI after they were discharged were excluded from the study. We performed a chi-squared test. Result A total of 132 patients were included in the study, of which 63 (47.7%) were males, while 69 (52.3%) were females. SSI was more commonly found among the age group of 41-70 years with 29 (38.7%) patients. Presurgical features including hemoglobin of >11 mg/dL, albumin of >3.5 g/dL, blood glucose of <80 mg/dL, and emergency surgery were found to be associated with the SSI having a significant p-value. Similarly, the surgical and postsurgical features significantly associated with the SSI having a significant p-value were the presence of surgical trauma, wound irrigation with normal saline, malignancy, bowel preparation, longer duration of the surgery, intraoperative hypotension, operative site, drain insertion, and the absence of a second antibiotic. Conclusion The early identification and management of the demographical, presurgical, surgical, and postsurgical risk factors can help reduce the incidence of SSIs. Bowel preparation should be encouraged, and unnecessary delays during the surgical process leading to increased procedure time should be avoided. Extra precaution needs to be provided for the patients highly susceptible to SSIs.


Copper (α)/cis-platinum-loaded nanogels as an adjuvant potentiate disulfiram's antitumor efficacy.
Disulfiram (DSF) is nontoxic and exerts anticancer activity by forming highly toxic chelates via its metabolite diethyldithiocarbamate with transition metal ions. However, there are not enough such ions in the human body to maintain the therapeutic effect. Herein, we report nanogels that complex copper ions or cis-platinum (CDDP) for tumor metal delivery to potentiate DSF's antitumor efficacy. We synthesized zwitterionic poly[N-(3-(methacryloyloxy-2-hydroxy)propyl)]-N-methyl glycine (PGMA-SAR) capable of chelating copper ions or CDDP and formed nanogels with suitable size and zeta potential. The intravenously injected nanogels circulated long in the blood compartment and delivered a high concentration of metal ions to the tumor. Separately administered DSF could sequester the metal ions from the nanogels and form highly cytotoxic complexes with potent in vitro and in vivo anticancer activity. This study provides a new strategy to potentiate DSF in anticancer treatment.


Reinforcement of Castor Oil-Based Polyurethane with Surface Modification of Attapulgite.
Polyurethane/attapulgite (PU/ATT) nanocomposites derived from castor oil were prepared by incorporation of 8 wt % ATT, acid-treated ATT, and KH560-treated ATT. The effects of three ATTs (ATT, acid-ATT, and KH560-ATT) on the comprehensive properties of PU/ATT nanocomposites were systematically investigated. The results showed that the incorporation of 8 wt % of three ATTs could produce an obvious reinforcement on the castor oil-based PU and that the silane modification treatment, rather than the acid treatment, has the more effective reinforcement effect. SEM images revealed the uniform dispersion of ATT in the PU matrix. DMA confirmed that the storage modulus and glass transition temperature (Tg) of PU/ATT nanocomposites were significantly increased after blending with different ATTs. For PU/KH560-ATT8 nanocomposites, the thermal stability of the PU was obviously enhanced by the addition of KH560-ATT. In particular, 8 wt % KH560-ATT loaded castor oil-based PU nanocomposites exhibit an obvious improvement in tensile strength (255%), Young's modulus (200%), Tg (5.1 °C), the storage modulus at 25 °C (104%), and the initial decomposition temperature (7.7 °C). The prepared bio-based PU materials could be a potential candidate to replace petroleum-based PU products in practical applications.


Spatiotemporal variation in vegetation spring phenology and its response to climate change in freshwater marshes of Northeast China.
Understanding wetland vegetation phenology and its response to climate change is important to predict the changes of wetland vegetation in wetland regions. Using the NDVI and climate data, this work studied the spatiotemporal change of start date of vegetation growing season (SOS) and explored the possible effects of climate change on the SOS over freshwater marshes of Northeast China. The results showed that the SOS significantly advanced by 0.52 day per year throughout the freshwater marshes of Northeast China during 2001 to 2016. The significant advancing of SOS was mainly concentrated in freshwater marshes of the Khingan Mountains (the Greater Khingan Mountains and the Lesser Khingan Mountains) and central arid or semi-arid regions (Songnen plain and Liaohe plain) in Northeast China. By contrast, there were weak delay trends of SOS in freshwater marshes of Eastern Inner Mongolia region, and Sanjiang plain. We found that precipitation was a dominant factor determining the SOS in arid or semi-arid regions (Songnen plain and Liaohe plain), while temperature played a bigger role in determining the SOS in Sanjiang plain and three cold mountains of the Northeast China. During the study period, increasing precipitation in the winter and spring contributed to advancing SOS in Songnen plain and Liaohe plain; the decrease of temperature from December to April explain the delaying SOS in freshwater marshes of Sanjiang Plain; the weak warming of temperature between November and May account for the advancing SOS of freshwater marshes in three cold mountains. In freshwater marshes of cold and the most arid region of Northeast China (Eastern Inner Mongolia), the SOS was influenced by both precipitation and temperature. Decreasing precipitation between January and April, as well as temperature decreases in March and April explain the delay of SOS in freshwater marshes of Eastern Inner Mongolia region.


Long-term persistence of translocations in stable lymphocytes from victims of a radiological accident.
To investigate whether translocations in 'stable' lymphocytes, i.e. those not containing unstable aberrations in any chromosome including counterstained ones, would have a longer persistence with time compared with those measured in all cells. The time-course of chromosomal aberrations in the three most highly exposed radiation victims of an Estonian accident in November 1994 was followed for 7 post-accident years encompassing 15 samples. Chromosome painting was performed using probes for chromosomes 1, 2 and 4 with a pan-centromeric probe, and chromosomal aberrations involving the painted chromosomes were scored using a developed version of the Protocol for Aberration Identification and Nomenclature Terminology (PAINT) nomenclature. Metaphases containing aberrations were captured with an image analyser and stored on a computer. An earlier analysis of aberrations in the painted portion of the genome was performed in all cells, irrespective of the possible aberrations in the unpainted part of the genome. The present analysis has taken into account the 'stable/unstable' nature of the complete cell. Evaluation was performed on images, counting all chromosomes and checking the counterstained chromosomes for unstable aberrations, i.e. dicentrics, acentrics or ring chromosomes. In the original analyses of all cells, a decrease in translocation frequency in the early samples was observed. In the present study of stable cells, the results showed that the yield of translocations is constant with time. The results show that translocations observed in stable cells are persistent with time. This implies that retrospective dosimetry and calibration should be performed using stable cells. To obtain more information on this issue, the stability status of all cells in any future fluorescence in situ hybridization follow-up of a radiation accident should be noted.


The clinical features and pathology of vasculitis associated with anti-myeloperoxidase autoantibodies.
Autoantibodies to myeloperoxidase (MPO) are associated with the autoimmune disease, systemic vasculitis, in humans. This results in severe inflammation and microscopic necrosis of multiple organs, especially the kidneys, leading to renal failure and death. The discovery of MPO autoantibodies has permitted the development of new diagnostic tests allowing earlier diagnosis and more effective therapy. Furthermore these antibodies are directly implicated in tissue injury by binding to MPO on the neutrophil cell membrane and stimulating neutrophil activation and degranulation. The causes for the breakdown in tolerance to MPO are not known although rare cases are drug-induced and remit on drug withdrawal. An understanding of the biology of MPO and its involvement in the pathogenesis of vasculitis is of importance in understanding the pathogenesis of vasculitis and the development of newer therapies.


[A PCR method for examining diarrhea--Escherichia coli].
To identify diarrhea--Escherichia coli(E. coli), especially Enteroinvasive E. coli(EIEC), Verocytotoxin-producing E. coli(VTEC), and Enterotoxigenic E. coli(ETEC), confirmation of pathogenic factors is necessary. We developed a system using the polymerase chain reaction(PCR) that simultaneously confirms the pathogenic genes of these E. coli, and have used it in routine examination. This method is simple and does not require examination for each pathogenic factor. Multiple pathogenic genes could be rapidly detected.


Growth and dissemination of Lewis lung carcinoma in plasminogen-deficient mice.
Plasminogen activation has been proposed to play a critical role in cancer invasion and metastasis. The effects of complete ablation of plasminogen activation in cancer was studied by inoculation of a metastatic Lewis lung carcinoma expressing high levels of plasminogen activator into plasminogen-deficient (Plg-/-) mice and matched control mice. Primary tumors developed in all mice with no difference in the rate of appearance between Plg-/- and control mice. However, the primary tumors in Plg-/- mice were smaller and less hemorrhagic and displayed reduced skin ulceration. In addition, dissemination of the tumor to regional lymph nodes was delayed in Plg-/- mice. Surprisingly, no quantitative differences were observed in lung metastasis between Plg-/- and control mice. In addition, Plg deficiency was compatible with metastasis of the primary tumor to a variety of other organs. Nevertheless, Plg-/- mice displayed a moderately increased survival after primary tumor resection. These findings suggest that plasmin-mediated proteolysis contributes to the morbidity and mortality of Lewis lung carcinoma in mice, but sufficient proteolytic activity is generated in Plg-/- mice for efficient tumor development and metastasis.


Convergent and Predictive Associations of Three Measures of Pedophilic Interest.
Pedophilic interest is a central risk factor for sexual offending against children. Multiple measures exist to assess pedophilic interest, and the present study aims to provide validity evidence for three of these measures in a sample of men convicted of sexual offenses. The association between a phallometric test for pedophilic interest, the Screening Scale for Pedophilic Interest (SSPI), and the sexual deviance factor of the Violence Risk Scale-Sexual Offense (VRS-SO) version was examined in a sample of 261 men who participated in sexual violence reduction services. The association between these measures and sexual recidivism, both as sole predictors and while controlling for static risk, was also assessed. The second aim of the study was to examine the validity of different methods for modeling the distribution of pedophilic interests, using phallometric test scores, based on the findings in recent taxometric research. The measures generally showed a positive and moderate relationship with each other and predicted sexual recidivism. However, the SSPI did not significantly predict sexual recidivism, and when controlling for static risk, only the VRS-SO Sexual Deviance factor significantly predicted this outcome. Modeling phallometric test scores continuously and trichotomously produced significant associations with sexual recidivism; however, only a trichotomous model with two levels remained predictive after controlling for static risk. The results are broadly supportive of measures of pedophilic interest and underscore the importance of appropriately modeling the latent structure of pedophilic interest.


Fatal outcome of jugular vein cannulation.
We describe the case of a woman who had cannulation of the internal jugular vein for the placement of a hemodialysis catheter. The procedure was unsuccessful and resulted in a retropharyngeal hematoma, which in turn caused severe compression on a carotid artery that was already severely narrowed by atherosclerosis. This resulted in massive cerebral hemispheric infarction and death. During the course of hospitalization, ultrasonography, computed tomography, and angiographic studies were done. Cannulation of neck vessels in patients with bleeding diatheses must be done with caution, especially in the presence of severe stenotic disease of the carotid arteries.


A dynamic DFT approach to generalized diffusion equations in a system with long-ranged and hydrodynamic interactions.
We build on an existing approximation scheme to the Smoluchowski equation in order to derive a dynamic density functional theory (DDFT) including two-body hydrodynamic interactions. A generalized diffusion equation and a wavenumber-dependent diffusion coefficient D(k) are derived by linearization in the density fluctuations. The result is applied to a colloidal monolayer at a fluid interface, having bulk-like hydrodynamic interactions and/or interacting via long-ranged capillary forces. In these cases, D(k) shows characteristic singularities as [Formula: see text]. The consequences of these singularities are studied by means of analytical perturbation theory, numerical solution of DDFT and simulations for an explicit example: the capillary collapse of a finite, disk-like distribution of particles. There is in general a good agreement between DDFT and simulations if the initial density distributions for the theoretical prediction correspond to the actual initial configurations of simulations, rather than to an average over them. Otherwise, discrepancies arise that are discussed in detail.


Estimation of the protein content of thyroid hormone receptor alpha 1 and beta 1 in rat tissues by western blotting.
Recent studies of the expression of c-erbA/thyroid hormone receptor (TR) mRNAs have revealed a dissociation between T3-binding activity and the behavior of the mRNAs that code the functional TRs in some tissues. Compared with T3-binding activity, TR(alpha 1 + beta 1) mRNA is disproportionally high in the brain and low in the liver. Using anti-TR antiserum, 4BII, which recognizes TR alpha 1 and beta 1, but not the alpha 2-variant, we measured TR protein content in rat tissues by Western blotting. Two protein bands of 47 and 55 kilodaltons (kDa) were specifically identified as TR proteins. The positions of the in vitro transcription/translation products of c-erbA/TR alpha 1 and beta 1 cDNA on the gel were consistent with those of the 47- and 55-kDa bands, respectively. The 47- and 55-kDa proteins in nuclear proteins extracted with 0.4 M KCl from rat tissues were analyzed by Western blotting, and the intensity of TR protein bands in each tissue was measured by a densitometer. The relative TR protein concentration was highest in liver, followed by brain, kidney, and testis. We compared the TR protein level measured by Western blotting with the maximal T3-binding capacity (Cmax) in the same aliquot of samples from liver and brain. Both the TR protein level and the Cmax in the brain were about 40% of those in the liver, suggesting that the Cmax per receptor molecule is constant in these two tissues, and an abundant amount of functional TR proteins exists in the liver, corresponding to the high level of T3-binding activity.


Expert Panel Recommendations for Use of Standardized Glucose Reporting System Based on Standardized Glucometrics Plus Visual Ambulatory Glucose Profile (AGP) Data in Clinical Practice.
This expert panel of diabetes specialists aimed to provide guidance to healthcare providers on the best practice in the use of innovative continuous glucose monitoring (CGM) techniques through a practical and implementable document that specifically addresses the rationale for and also analysis and interpretation of the new standardized glucose reporting system based on standardized CGM metrics and visual ambulatory glucose profile (AGP) data. This guidance document presents recommendations and a useful algorithm for the use of a standardized glucose reporting system in the routine diabetes care setting.


Micro-computed tomographic assessment of the shaping ability of the One Curve, One Shape, and ProTaper Next nickel-titanium rotary systems.
This micro-computed tomographic (CT) study aimed to compare the shaping abilities of ProTaper Next (PTN), One Shape (OS), and One Curve (OC) files in 3-dimensionally (3D)-printed mandibular molars. In order to ensure standardization, 3D-printed mandibular molars with a consistent mesiobuccal canal curvature (45°) were used in the present study (n = 18). Specimens were instrumented with the OC, OS, or PTN files. The teeth were scanned pre- and post-instrumentation using micro-CT to detect changes of the canal volume and surface area, as well as to quantify transportation of the canals after instrumentation. Two-way analysis of variance was used for statistical comparisons. No statistically significant differences were found between the OC and OS groups in the changes of the canal volume and surface area before and after instrumentation (p > 0.05). The OC files showed significantly less transportation than the OS or PTN systems for the apical section (p < 0.05). In a comparison of the systems, similar values were found at the coronal and middle levels, without any significant differences (p > 0.05). These 3 instrumentation systems showed similar shaping abilities, although the OC file achieved a lesser extent of transportation in the apical zone than the OS and PTN files. All 3 file systems were confirmed to be safe for use in mandibular mesial canals.


Intradermal pre-exposure rabies immunisation in New Zealand.
Rabies is a fatal infection and immunisation is important to consider in those travellers going to rabies endemic areas. In those at high risk, a course of three immunisations may be given by the intramuscular (IM) or intradermal (ID) route, both of which are approved by the World Health Organization (WHO) and the Centers for Disease Control (CDC). Little is known in the New Zealand context regarding the effectiveness of pre-exposure ID rabies immunisation. The data was collected prospectively on all travellers requiring the immunisation from July 2001 to September 2003 in Auckland. The standard WHO rabies immunisation protocol was used with three ID injections of 0.1 ml, given on days 0, 7, and 21 or 28 with a booster after 12 months. The vaccine used was the Pasteur Merieux human diploid cell vaccine (HDCV) or the Rabipur Purified chick embryo cell (PCEC) vaccine. Both vaccines are approved by the WHO and the CDC, and are interchangeable. Serology was performed approximately 2 weeks after completion of the primary immunisation course or after a booster, wherever possible. Antibody levels were measured using EIA, and levels of >0.5 IU/ml were considered protective. Of the 263 travellers assessed in this study, 125 were males and 138 were females. The mean age of the cohort was 34.8 years (SD=11.7). There were not found to be any statistically significant correlations between age and antibody levels neither was there any significant association between gender and antibody levels. In addition to the sample group, a further 12 travellers had rabies serology performed but were excluded from the study because they had IM vaccines as part of their primary course. Whilst rabies serology ranged from 0.2 to 27.9 IU/ml in the study cohort, the mean antibody level for the group was 4.7 IU/ml (SD=4.1 IU/ml). The mean antibody level for males was 4.3 IU/ml (SD=3.3), and for females, 5.2 IU/ml (SD=4.6). Of the 263 travellers, all had some level of detectable antibodies. The overall seroconversion rate was 95.1%. ID rabies immunisation appears effective, when given according to the standard WHO protocol, in New Zealand. ID rabies immunisation is also more affordable for travellers, especially those on a restrictive budget. ID rabies immunisation can continue to be recommended, particularly where follow-up serology can be done before travel and where there are staff who are experienced in ID immunisation.


Molecular characterization of Iranian wheat stripe virus shows its taxonomic position as a distinct species in the genus Tenuivirus.
The full lengths of three genome segments of Iranian wheat stripe virus (IWSV) were amplified by reverse transcription (RT) followed by polymerase chain reaction (PCR) using a primer complementary to tenuivirus conserved terminal sequences. The segments were sequenced and found to comprise 3469, 2337, and 1831 nt, respectively. The gene organization of these segments is similar to that of other known tenuiviruses, each displaying an ambisense coding strategy. IWSV segments, however, are different from those of other viruses with respect to the number of nucleotides and deduced amino acid sequence for each ORF. Depending on the segment, the first 16-22 nt at the 5' end and the first 16 nt at the 3' end are highly conserved among IWSV and rice hoja blanca virus (RHBV), rice stripe virus (RSV) and maize stripe virus (MStV). In addition, the first 15-18 nt at the 5' end are complementary to the first 16-18 nt at the 3' end. Phylogenetic analyses showed close similarity and a common ancestor for IWSV, RHBV, and Echinochloa hoja blanca virus (EHBV). These findings confirm the position of IWSV as a distinct species in the genus Tenuivirus.


A DESCRIPTION OF ARTERIAL BLOOD PRESSURE MEASUREMENT IN TWO SPECIES OF FLYING FOXES (PTEROPUS VAMPYRUS AND PTEROPUS HYPOMELANUS).
Blood pressure assessment is valuable during management of chronic conditions with increased risk of developing hypertension and as a standard practice for anesthetic monitoring. Normal arterial blood pressure values have not been well described in megachiropteran species. Following anesthetic induction and maintenance with isoflurane in oxygen, arterial blood pressure was obtained from the posterior tibial artery of eight large flying foxes (Pteropus vampyrus) and six variable flying foxes (Pteropus hypomelanus), two with structural cardiac disease and four in good clinically health. Normal values reported as a median with interquartile range for systolic, diastolic, and mean (MAP) arterial pressures for P. vampyrus were 101 (94, 107), 69 (57, 80), and 86 (75, 93), respectively. Normal MAP for clinically healthy P. hypomelanus was 86 (67, 93). Placement of P. hypomelanus in a vertical head-down position did not alter blood pressure in clinically healthy bats, but significantly increased MAP in two bats with structural cardiac disease. Arterial catheterization of both the posterior tibial and median arteries in these species was easily performed without major complication.


The effect of 2 weeks vitamin C supplementation on immunoendocrine responses to 2.5 h cycling exercise in man.
An increased systemic concentration of stress hormones (of the hypothalamic-pituitary adrenal axis) and some cytokines may contribute to the depression of immune cell function typically observed after prolonged exercise. The aim of the present study was to determine the effect of 2 weeks of supplementation with vitamin C (VC) on cortisol, adrenocorticotrophic hormone, interleukin-6, oxidative stress and neutrophil responses to a single bout of endurance exercise. Nine healthy endurance-trained males exercised for 2.5 h at 60% VO2max after 2 weeks of placebo (PLA) or VC (1,000 mg day(-1)) supplementation. All participants completed both trials utilising a randomised crossover design with a minimum 14 day washout period between trials. There was a significant trial x time interaction effect for plasma cortisol concentration (P = 0.039) which tended to be lower in the VC trial but post hoc analysis found no specific between trial differences. There was a significantly lower post-exercise neutrophilia (P < 0.014) in the VC trial, compared with the PLA trial. There was no trial x time interaction for measures of neutrophil function (bacteria-stimulated elastase release, fMLP or PMA-stimulated oxidative burst). However, there was a trend for higher fMLP-stimulated neutrophil oxidative burst in the VC compared with PLA trial (trial x time interaction, P = 0.075). These results suggest that supplementation with VC for a period of up to 2 weeks provides little to no protection against the depression of neutrophil function which typically occurs after endurance exercise.


Detection of Helicobacter pylori infections in patients with gastric syndrome.
Results of microbiological and serological studies, as well as QUT (quick urease test) test done for 204 patients with upper gastrointestinal tract's complaints are presented in this paper. Microbiological studies and QUT test were done with mucous membrane biopsies, taken during gastroscopy or during surgery. Serological studies were performed triple on commercial latex tests and on ELISA-test. Results obtained by QUT test, microbiological and serological studies were comparable. ELISA-test can be recommended as reference test for detection of H. pylori infections.


p28 Bacterial Peptide, as an Anticancer Agent.
Cancer remains a major cause of morbidity and mortality irrespective of the type of conventional chemotherapy. Therefore, there is an urgent need for new and effective anticancer therapeutic agents. Bacterial proteins and their derivative peptides appear as a promising approach for cancer treatment. Several, including an amphipathic, α-helical, 28-amino acid peptide derived from azurin, a 128-amino acid copper-containing redox protein secreted from Pseudomonas aeruginosa, show clinical promise in the treatment of adult and pediatric solid tumors. The peptide, p28, is a post-translational, multi-target anticancer agent that preferentially enters a wide variety of solid tumor cells. Mechanistically, after entry, p28 has two major avenues of action. It binds to both wild-type and mutant p53 protein, inhibiting constitutional morphogenic protein 1 (Cop1)-mediated ubiquitination and proteasomal degradation of p53. This results in increased levels of p53, which induce cell-cycle arrest at G2/M and an eventual apoptosis that results in tumor cell shrinkage and death. In addition, p28 also preferentially enters nascent endothelial cells and decreases the phosphorylation of FAK and Akt inhibiting endothelial cell motility and migration. Here, we review the current basic and clinical evidence suggesting the potential of p28 as a cancer therapeutic peptide.


Voluntary muscle function after creatine supplementation in acute hypobaric hypoxia.
To determine whether creatine (Cr) supplementation improves muscle performance during exposure to acute hypobaric hypoxia. Seven healthy men (28 +/- 6 yr, mean +/- SD) performed submaximal intermittent static knee contractions interspersed with maximal voluntary contractions (MVCs) every minute to exhaustion (approximately 50% of rested MVC force) in normoxia and hypobaric hypoxia (separated by 3 d) after supplementation with Cr (20 g.d(-1) for 7 d then 5 g.d(-1) for 4-7 d) or placebo (Pla) in a double-blind, randomized crossover study. A 5-wk period without supplementation separated treatments. Each test day, subjects performed two bouts (separated by 2 min) at their preset submaximal force, 32 +/- 4% rested MVC). Rested MVC force (860 +/- 66 N) and MVC force at exhaustion (396 +/- 27 N; 47 +/- 3% rested MVC) did not differ among treatments or environments (P > 0.05). For bout 1, endurance time was shorter in hypobaria (26 +/- 3 min) than normoxia (34 +/- 2 min) (P < 0.01), but did not differ between Cr (27 +/- 3 min) and Pla (33 +/- 3 min) (P > 0.05). MVC force returned to similar levels (P >0.05) in bout 2 after recovery in all four sessions (to approximately 615 N). For bout 2, endurance time also was shorter in hypobaria (7 +/- 1 min) than normoxia (9 +/- 1 min) (P < 0.03) but did not differ between Cr and Pla (P > 0.05). This study, which used an exercise model designed to impose the same target contraction force under all experimental conditions, found no effect of Cr on maximal force, muscle endurance, or recovery in normoxia or hypobaric hypoxia.


Dissecting the suicide phenotype: the role of impulsive-aggressive behaviours.
Suicide ranks among the main causes of death in most Western countries and is, thus, an important public health problem. Over the last few years, it has been increasingly recognized that people who attempt or commit suicide have a certain individual predisposition, partly resulting from certain personality traits, in particular, impulsive-aggressive behaviors. The evidence suggests that impulsive-aggressive traits are part of a developmental cascade that increases suicide risk among a subset of suicides. These personality traits may also mediate familial transmission of suicidal behavior, defining a behavioral endophenotype that could be useful in molecular genetic studies of suicide. On the other hand, not all suicides are associated with impulsive-aggressive behaviors. There is a need to better understand the mechanisms mediating suicide among individuals with no history of impulsive-aggressive behaviors.


Pediatric dosing--the pharmacist's dilemma.
Several pediatric dosing rules commonly are published in medical and pharmaceutical texts. However, most authorities do not regard these published dosing rules (e.g., Clark's weight rule) to be responsible approaches to pediatric dosing. This study compares the actual doses administered to patients in a 310-bed, university-affiliated, pediatric hospital with the dose calculated for each patient using Clark's weight rule, Clark's surface area rule, Young's age rule, and Shirkey's dosing recommendations. A t test analysis for paired comparisons revealed that the actual dose administered differed significantly from doses calculated, except for doses calculated using Clark's surface area rule. The dosage deviation range was greatest for Clark's surface area rule. This suggests that Clark's surface area rule is also unreliable. Responsible dosing must involve more than the application of commonly published, pediatric dosing calculation rules. Shirkey's recommendations provided the most reasonable approach to pediatric dosing. However, pharmacists should monitor these patients carefully and recommend dosage adjustments whenever necessary.


Hemolysis and Plasma Free Hemoglobin During Extracorporeal Membrane Oxygenation Support: From Clinical Implications to Laboratory Details.
Venovenous and venoarterial extracorporeal membrane oxygenation (ECMO) are lifesaving supports that are more and more frequently used in critically ill patients. Despite of major technological improvements observed during the last 20 years, ECMO-associated hemolysis is still a complication that may arise during such therapy. Hemolysis severity, directly appreciated by plasma free hemoglobin concentration, may be present with various intensity, from a nonalarming and tolerable hemolysis to a highly toxic one. Here, we propose a review dedicated to extracorporeal membrane oxygenation (ECMO)-associated hemolysis, with a particular emphasis on pathophysiology, prevalence, and clinical consequences of such complication. We also focus on laboratory assessment of hemolysis and on the limits that have to be known by clinicians to prevent and manage hemolytic events.


The effect of D2343 in transmurally stimulated rabbit isthmus muscle.
In in vitro experiments spirally cut isthmic preparations from oestrogenized rabbits have been contracted by transmural stimulation. The beta 2-agonistic and alpha 1-antagonistic drug D2343 (for chemical structure, see fig. 1 in Olsson et al. 1984) inhibits the evoked contractions in a concentration related fashion with 50% effect at 9.6 X 10(-6) mol/l. The alpha 1-antagonist prazosin has an inhibiting action in the same concentration range (EC50 = 14.0 X 10(-6) mol/l). To inhibit the contractile response to the same degree by the beta 2-agonist terbutaline the concentration has to be greater than 10(-3) mol/l. From received results the beta (beta 2)-adrenoceptor mediated effects seem to be of minor importance compared to the alpha (alpha 1)-receptor dominance in the isthmus muscle during oestrus.


Growth arrest-specific protein 2 (GAS2) interacts with CXCR4 to promote T-cell leukemogenesis partially via c-MYC.
Although growth arrest-specific protein 2 (GAS2) promotes the growth of T-cell acute lymphoblastic leukemia (T-ALL) cells in culture, the effect of GAS2 on T-cell leukemogenesis has not been studied, and the mechanism remains unclear. In the present study, xenograft studies showed that GAS2 silencing impaired T-cell leukemogenesis and decreased leukemic cell infiltration. Mechanistically, GAS2 regulated the protein expression of C-X-C chemokine receptor type 4 (CXCR4) rather than its transcript expression. Immunoprecipitation revealed that GAS2 interacted with CXCR4, and confocal analysis showed that GAS2 was partially co-expressed with CXCR4, which provided a strong molecular basis for GAS2 to regulate CXCR4 expression. Importantly, CXCR4 overexpression alleviated the inhibitory effect of GAS2 silencing on the growth and migration of T-ALL cells. Moreover, GAS2 or CXCR4 silencing inhibited the expression of NOTCH1 and c-MYC. Forced expression of c-MYC rescued the growth suppression induced by GAS2 or CXCR4 silencing. Meanwhile, GAS2 deficiency, specifically in blood cells, had a mild effect on normal hematopoiesis, including T-cell development, and GAS2 silencing did not affect the growth of normal human CD3+ or CD34+ cells. Overall, our data indicate that GAS2 promotes T-cell leukemogenesis through its interaction with CXCR4 to activate NOTCH1/c-MYC, whereas impaired GAS2 expression has a mild effect on normal hematopoiesis. Therefore, our study suggests that targeting the GAS2/CXCR4 axis is a potential therapeutic strategy for T-ALL.


In vivo tissue regeneration with robotic implants.
Robots that reside inside the body to restore or enhance biological function have long been a staple of science fiction. Creating such robotic implants poses challenges both in signaling between the implant and the biological host, as well as in implant design. To investigate these challenges, we created a robotic implant to perform in vivo tissue regeneration via mechanostimulation. The robot is designed to induce lengthening of tubular organs, such as the esophagus and intestines, by computer-controlled application of traction forces. Esophageal testing in swine demonstrates that the applied forces can induce cell proliferation and lengthening of the organ without a reduction in diameter, while the animal is awake, mobile, and able to eat normally. Such robots can serve as research tools for studying mechanotransduction-based signaling and can also be used clinically for conditions such as long-gap esophageal atresia and short bowel syndrome.


Xanthine oxidase activation in mild gestational hypertension.
We hypothesized that activation of the xanthine oxidase (XO) enzyme system is a potential source of free radicals in pregnancy-induced hypertension (PIH). A prospective observational study was carried out on 16 pregnant women who met the criteria of gestational hypertension [rise in blood pressure (BP) of 30 mm Hg systolic or 15 mm Hg diastolic after 20 weeks gestation or BP>140/90 mm Hg if earlier pressure is unknown] without proteinuria or any signs of renal impairment. Fourteen women with a clinically normal pregnancy matched for maternal age, parity, and gestational age acted as pregnant controls. Nonpregnant control women were members of the laboratory staff ( n=15). Concentrations of free sulfhydryl (SH) groups, purine catabolites, lipid peroxidation products in plasma, and blood carboxyhemoglobin levels were used to follow oxidative stress and potential hemolysis. A noninvasive measurement of functional XO activity was carried out (i.e., the urinary ratio of the two metabolites of caffeine was estimated). A pronounced oxidative stress was demonstrated in plasma samples of patients with hypertension by the elevated concentrations of uric acid and lipid peroxidation products. A reduced level of free sulfhydryl groups and an increased concentration of hypoxanthine (HX) were shown in normotensive pregnant individuals. The XO activity index was substantially higher in overweight pregnant subjects with mild hypertension [0.849+/-0.096 ( p<0.01)] than in normotensive pregnant women or in age-matched nonpregnant subjects [0.596+/-0.105, 0.542+/-0.049 (means+/-SD), respectively]. Our study of mildly hypertensive pregnant subjects provides additional evidence of the putative role of XO activation as a source of free radicals in the early stage of endothelial dysfunction.


Modifying an Active Compound's Release Kinetic Using a Supercritical Impregnation Process to Incorporate an Active Agent into PLA Electrospun Mats.
The main objective of this work was to study the release of cinnamaldehyde (CIN) from electrospun poly lactic acid (e-PLA) mats obtained through two techniques: (i) direct incorporation of active compound during the electrospinning process (e-PLA-CIN); and (ii) supercritical carbon dioxide (scCO₂) impregnation of CIN within electrospun PLA mats (e-PLA/CINimp). The development and characterization of both of these active electrospun mats were investigated with the main purpose of modifying the release kinetic of this active compound. Morphological, structural, and thermal properties of these materials were also studied, and control mats e-PLA and e- PLA CO 2 were developed in order to understand the effect of electrospinning and scCO₂ impregnation, respectively, on PLA properties. Both strategies of incorporation of this active compound into PLA matrix resulted in different morphologies that influenced chemical and physical properties of these composites and in different release kinetics of CIN. The electrospinning and scCO₂ impregnation processes and the presence of CIN altered PLA thermal and structural properties when compared to an extruded PLA material. The incorporation of CIN through scCO₂ impregnation resulted in higher release rate and lower diffusion coefficients when compared to active electrospun mats with CIN incorporated during the electrospinning process.


Codon bias analyses on thyroid carcinoma genes.
Thyroid carcinoma is one of the most common cancers in the world. Although the genetics of thyroid carcinoma was intensively studied, new mechanisms could be involved in its development as the codon bias. In this paper, we studied the codon bias of thyroid-cancer genes, considering not only the sequences but also the synonymous mutations. Different measures and statistical analyses were employed to characterize the thyroid-cancer genes. We considered classical measures as RSCU and ENC, the compositional and protein characteristics, but also the codon bias landscape via the %MinMax algorithm. The compositional analyses highlighted two groups of thyroid cancer genes according to the GC% and GC3% content. The ENC did not show a clear codon bias in the genes. Differently, the RSCU analyses showed interesting codons that could play an important role in the development of thyroid cancer as the codon Ser-tcG. Furthermore, interesting synonymous mutations were detected that could affect the codon bias. The codon bias landscape detected genes enriched in rare codons as AKAP9 and KTN1. A cluster analysis based on %MinMax classified the thyroid cancer genes in four different groups according to the distribution of rare/frequent codons in the sequence. This is the first study that analyzed the codon bias in thyroid cancer genes based also on synonymous mutations. This study provided different hints that should be further investigated by wet-lab validation and that it could open new scenarios in the understanding the molecular mechanisms involved in thyroid cancer development based on codon bias.


Polygenic risk scores across the extended psychosis spectrum.
As early detection of symptoms in the subclinical to clinical psychosis spectrum may improve health outcomes, knowing the probabilistic susceptibility of developing a disorder could guide mitigation measures and clinical intervention. In this context, polygenic risk scores (PRSs) quantifying the additive effects of multiple common genetic variants hold the potential to predict complex diseases and index severity gradients. PRSs for schizophrenia (SZ) and bipolar disorder (BD) were computed using Bayesian regression and continuous shrinkage priors based on the latest SZ and BD genome-wide association studies (Psychiatric Genomics Consortium, third release). Eight well-phenotyped groups (n = 1580; 56% males) were assessed: control (n = 305), lower (n = 117) and higher (n = 113) schizotypy (both groups of healthy individuals), at-risk for psychosis (n = 120), BD type-I (n = 359), BD type-II (n = 96), schizoaffective disorder (n = 86), and SZ groups (n = 384). PRS differences were investigated for binary traits and the quantitative Positive and Negative Syndrome Scale. Both BD-PRS and SZ-PRS significantly differentiated controls from at-risk and clinical groups (Nagelkerke's pseudo-R2: 1.3-7.7%), except for BD type-II for SZ-PRS. Out of 28 pairwise comparisons for SZ-PRS and BD-PRS, 9 and 12, respectively, reached the Bonferroni-corrected significance. BD-PRS differed between control and at-risk groups, but not between at-risk and BD type-I groups. There was no difference between controls and schizotypy. SZ-PRSs, but not BD-PRSs, were positively associated with transdiagnostic symptomology. Overall, PRSs support the continuum model across the psychosis spectrum at the genomic level with possible irregularities for schizotypy. The at-risk state demands heightened clinical attention and research addressing symptom course specifiers. Continued efforts are needed to refine the diagnostic and prognostic accuracy of PRSs in mental healthcare.


Sphingosine-1-phosphate prevents permeability increases via activation of endothelial sphingosine-1-phosphate receptor 1 in rat venules.
Sphingosine-1-phosphate (S1P) has been demonstrated to enhance endothelial barrier function in vivo and in vitro. However, different S1P receptor subtypes have been indicated to play different or even opposing roles in the regulation of vascular barrier function. This study aims to differentiate the roles of endogenous endothelial S1P subtype receptors in the regulation of permeability in intact microvessels using specific receptor agonist and antagonists. Microvessel permeability was measured with hydraulic conductivity (L(p)) in individually perfused rat mesenteric venules. S1P-mediated changes in endothelial intracellular Ca(2+) concentration ([Ca(2+)](i)) was measured in fura-2-loaded venules. Confocal images of fluorescent immunostaining illustrated the spatial expressions of three S1P subtype receptors (S1P(R1-3)) in rat venules. The application of S1P (1 μM) in the presence of S1P(R1-3) inhibited platelet-activating factor- or bradykinin-induced permeability increase. This S1P effect was reversed only with a selective S1P(R1) antagonist, W-146, and was not affected by S1P(R2) or S1P(R3) antagonists JTE-013 and CAY-10444, respectively. S1P(R1) was also identified as the sole receptor responsible for S1P-mediated increases in endothelial [Ca(2+)](i). S1P(R2) or S1P(R3) antagonist alone affected neither basal L(p) nor platelet-activating factor-induced permeability increase. The selective S1P(R1) agonist, SEW-2871, showed similar [Ca(2+)](i) and permeability effect to that of S1P. These results indicate that, despite the presence of S1P(R1-3) in the intact venules, only the activation of endothelial S1P(R1) is responsible for the protective action of S1P on microvessel permeability and that endogenous S1P(R2) or S1P(R3) did not exhibit functional roles in the regulation of permeability under basal or acutely stimulated conditions.


Exhibition of anti-platelet effect of isocarbacyclin methyl ester and its free acid via an increase in cAMP in platelets.
Isocarbacyclin methyl ester (CAS 88931-51-5, TEI-9090) and its free acid (CAS 88911-35-7, TEI-7165) are chemically stable analogues of prostaglandin (PG) I2. The cAMP increasing activities of TEI-9090 and TEI-7165 were investigated. TEI-9090, TEI-7165, PGI2, and PGE1 caused the accumulation of cAMP in rabbit and human platelets. The effects of TEI-9090, TEI-7165, and PGE1 were longer lasting than the effect of PGI2. Each drug concentration-dependently increased the cAMP level in rabbit and human platelets. The order of cAMP-increasing activity was PGI2 > PGE1 = TEI-9090 = TEI-7165 in rabbit platelets and PGI2 > TEI-9090 = TEI-7165 > PGE1 in human platelets. The antiplatelet aggregation activity of TEI-9090 in rabbit and human platelets was remarkably decreased by the addition of an esterase inhibitor, diisopropyl fluorophosphate (DFP). However, this activity of TEI-7165 was not affected by the inhibitor. In the presence of DFP, the cAMP accumulation by TEI-9090 in rabbit platelets was almost completely inhibited but that by TEI-7165 was not inhibited. These results suggest that TEI-9090 is deesterified to TEI-7165 and that this free acid then inhibits platelet aggregation via an increase in the cAMP level in the platelets.


Anatomic and physiologic approach for trans-conjunctival needle decompression of orbital emphysema.
A 19-year-old with blunt trauma and repeated nose blowing presented with orbital emphysema and orbital compartment syndrome. Orbital emphysema is the abnormal presence of air within the orbit, typically secondary to trauma. Most cases will resolve with observation alone, however orbital compartment syndrome is a feared complication that necessitates urgent decompression. A superior fornix, trans-conjunctival approach was safely utilized to decompress the orbit while avoiding unwanted complications. Various decompression techniques have previously been described, most of which describe trans-palpebral approaches. The key safety benefit to the trans-conjunctival approach is direct visualization of the needle tip adjacent to the superior fornix, therefore posterior to the equator of the globe. With the needle positioned parallel to the curvature of the globe at the equator, the posterior sclera surface curves away from the tip, rendering it difficult to pierce the globe. In addition, the needle needs to be advanced only 3-4 millimeters and traverses only the conjunctiva and Tenon's to enter the central surgical space - the most direct route to the air pocket. The trans-conjunctival approach lowers the risk of damage to eyelid neurovascular structures and is less painful.


The Stockholm Neonatal Family Centered Care Study: effects on length of stay and infant morbidity.
Parental involvement in the care of preterm infants in NICUs is becoming increasingly common, but little is known about its effect on infants' length of hospital stay and infant morbidity. Our goal was to evaluate the effect of a new model of family care (FC) in a level 2 NICU, where parents could stay 24 hours/day from admission to discharge. A randomized, controlled trial was conducted in 2 NICUs (both level 2), including a standard care (SC) ward and an FC ward, where parents could stay from infant admission to discharge. In total, 366 infants born before 37$$\raisebox{1ex}{$0$}\!\left/ \!\raisebox{-1ex}{$7$}\right.$$ weeks of gestation were randomly assigned to FC or SC on admission. The primary outcome was total length of hospital stay, and the secondary outcome was short-term infant morbidity. The analyses were adjusted for maternal ethnic background, gestational age, and hospital site. Total length of hospital stay was reduced by 5.3 days: from a mean of 32.8 days (95% confidence interval [CI]: 29.6-35.9) in SC to 27.4 days (95% CI: 23.2-31.7) in FC (P = .05). This difference was mainly related to the period of intensive care. No statistical differences were observed in infant morbidity, except for a reduced risk of moderate-to-severe bronchopulmonary dysplasia: 1.6% in the FC group compared with 6.0% in the SC group (adjusted odds ratio: 0.18 [95% CI: 0.04-0.8]). Providing facilities for parents to stay in the neonatal unit from admission to discharge may reduce the total length of stay for infants born prematurely. The reduced risk of moderate-to-severe bronchopulmonary dysplasia needs additional investigation.


How physical forces drive the process of helical membrane protein folding.
Protein folding is a fundamental process of life with important implications throughout biology. Indeed, tens of thousands of mutations have been associated with diseases, and most of these mutations are believed to affect protein folding rather than function. Correct folding is also a key element of design. These factors have motivated decades of research on protein folding. Unfortunately, knowledge of membrane protein folding lags that of soluble proteins. This gap is partly caused by the greater technical challenges associated with membrane protein studies, but also because of additional complexities. While soluble proteins fold in a homogenous water environment, membrane proteins fold in a setting that ranges from bulk water to highly charged to apolar. Thus, the forces that drive folding vary in different regions of the protein, and this complexity needs to be incorporated into our understanding of the folding process. Here, we review our understanding of membrane protein folding biophysics. Despite the greater challenge, better model systems and new experimental techniques are starting to unravel the forces and pathways in membrane protein folding.


Participation of chlorophyll b reductase in the initial step of the degradation of light-harvesting chlorophyll a/b-protein complexes in Arabidopsis.
The light-harvesting chlorophyll a/b-protein complex of photosystem II (LHCII) is the most abundant membrane protein in green plants, and its degradation is a crucial process for the acclimation to high light conditions and for the recovery of nitrogen (N) and carbon (C) during senescence. However, the molecular mechanism of LHCII degradation is largely unknown. Here, we report that chlorophyll b reductase, which catalyzes the first step of chlorophyll b degradation, plays a central role in LHCII degradation. When the genes for chlorophyll b reductases NOL and NYC1 were disrupted in Arabidopsis thaliana, chlorophyll b and LHCII were not degraded during senescence, whereas other pigment complexes completely disappeared. When purified trimeric LHCII was incubated with recombinant chlorophyll b reductase (NOL), expressed in Escherichia coli, the chlorophyll b in LHCII was converted to 7-hydroxymethyl chlorophyll a. Accompanying this conversion, chlorophylls were released from LHCII apoproteins until all the chlorophyll molecules in LHCII dissociated from the complexes. Chlorophyll-depleted LHCII apoproteins did not dissociate into monomeric forms but remained in the trimeric form. Based on these results, we propose the novel hypothesis that chlorophyll b reductase catalyzes the initial step of LHCII degradation, and that trimeric LHCII is a substrate of LHCII degradation.


[Optic neuropathy of dental origin. Apropos of 4 cases].
The optic neuropathy due to dental affection is rare but functionally serious. Authors report four cases of patients who have an acute dental infection with unilateral inflammation of the optic nerve. The first symptom was in all cases a sudden fall of visual acuity. The visual field is always altered. Papilloedema is observed in three eyes. The treatment is of the dental infection at first, secondly, corticosteroïds are associated. An improvement is noticed in three cases. The authors discuss the physiopathology of the affection, its symptoms and treatment.


Typical curve with G1 constraints for curve completion.
This paper presents a novel algorithm for planar G1 interpolation using typical curves with monotonic curvature. The G1 interpolation problem is converted into a system of nonlinear equations and sufficient conditions are provided to check whether there is a solution. The proposed algorithm was applied to a curve completion task. The main advantages of the proposed method are its simple construction, compatibility with NURBS, and monotonic curvature.


[T cell factor 3 (TCF3) is overexpressed in hepatocellular carcinoma and promotes their invasion and metastasis].
Objective To investigate the expression of T cell factor 3 (TCF3) in hepatocellular carcinoma (HCC), its correlation with the prognosis of HCC patients, and its effect on the invasion, migration, and metastasis of HCC cells. Methods The expression of TCF3 mRNA in HCC tissues was detected with tumor public databases and the expression of TCF3 protein in HCC specimens was detected by immunohistochemical staining. Correlation between TCF3 expression and HCC patients' prognosis was analyzed. Western blot analysis was used to detect the expression of TCF3 in different human HCC cell lines, and lentivirus infection was conducted to construct TCF3-upregulated and TCF3-downregulated HCC cell lines. The effect of TCF3 on the invasion and migration of HCC cells was assessed by in vitro TranswellTM assay, and in vivo intrahepatic tumor implantation models were established to evaluate the effect of TCF3 on the metastatic capacity of HCC cells. Results The expression of TCF3 mRNA was significantly higher in HCC tissues than that in normal liver tissues, and high expression of TCF3 mRNA was closely correlated with decreased overall survival rates of HCC patients. In 120 cases of HCC tissues, the protein level of TCF3 was significantly higher than that in adjacent nontumor tissues, and patients with positive TCF3 expression had a markedly decreased overall survival rate and a higher recurrence rate compared with patients with negative TCF3 expression. In vitro TranswellTM assay indicated that TCF3 upregulation promoted the invasion and migration of PLC/PRF/5 cells, whereas knockdown of TCF3 inhibited the invasion and migration abilities of HCCLM3 cells. Intrahepatic tumor implantation models showed that TCF3 upregulation promoted the metastasis of PLC/PRF/5 cells, while TCF3 knockdown weakened the metastatic capacity of HCCLM3 cells. Conclusion TCF3 expression is significantly upregulated in human HCC tissues, and high TCF3 expression predicts a poor prognosis of HCC patients. TCF3 markedly promotes the invasion, migration, and metastasis of HCC cells.