Pathology of the ear in murine mucopolysaccharidosis type VII. Morphologic correlates of hearing loss.
Patients with mucopolysaccharidosis commonly have hearing impairment but the morphologic alterations in the ear caused by these lysosomal storage diseases are incompletely defined. We studied a murine model of mucopolysaccharidosis VII with clinical features, including conductive hearing loss and biochemical, and pathologic features similar to those seen in human mucopolysaccharidoses. Gross morphology, radiography, light and electron microscopy were used to define the pathologic alterations in the ear that correlate with auditory dysfunction in mucopolysaccharidosis VII. Cerumen occluded the external auditory canal and there was a severe otitis media. The bone encasing the middle and inner ear was sclerotic and opaque and the temporal bone and the ossicles and their joints contained cells distended by enlarged lysosomes. Hair cell damage and multifocal lysosomal distention in endoneural fibroblasts and spiral ganglion neurons characterized the mucopolysaccharidosis VII cochlea. The external auditory canal obstruction, otitis media, and ossicle articular alterations in mucopolysaccharidosis VII mice cause a conductive hearing loss. The hair cell damage and neuronal storage may contribute to sensorineural deafness. This model allows investigation of the pathophysiology of auditory dysfunction in mucopolysaccharidosis and the effects of therapies on hearing loss.


Real-Time Cameraless Measurement System Based on Bioelectrical Ventilatory Signals to Evaluate Fear and Anxiety.
Fear and anxiety in fish are generally evaluated by video-based behavioral analysis. However, it is difficult to distinguish the psychological state of fish exclusively through video analysis, particularly whether the fish are freezing, which represents typical fear behavior, or merely resting. We propose a system that can measure bioelectrical signals called ventilatory signals and simultaneously analyze swimming behavior in real time. Experimental results comparing the behavioral analysis of the proposed system and the camera system showed a low error level with an average absolute position error of 9.75 ± 3.12 mm (about one-third of the body length) and a correlation between swimming speeds of r = 0.93 ± 0.07 (p < 0.01). We also exposed the fish to zebrafish skin extracts containing alarm substances that induce fear and anxiety responses to evaluate their emotional changes. The results confirmed that this solution significantly changed all behavioral and ventilatory signal indices obtained by the proposed system (p < 0.01). By combining the behavioral and ventilatory signal indices, we could detect fear and anxiety with a discrimination rate of 83.3% ± 16.7%. Furthermore, we found that the decreasing fear and anxiety over time could be detected according to the peak frequency of the ventilatory signals, which cannot be measured through video analysis.


Treatment of charge singularities in implicit solvent models.
This paper presents a novel method for solving the Poisson-Boltzmann (PB) equation based on a rigorous treatment of geometric singularities of the dielectric interface and a Green's function formulation of charge singularities. Geometric singularities, such as cusps and self-intersecting surfaces, in the dielectric interfaces are bottleneck in developing highly accurate PB solvers. Based on an advanced mathematical technique, the matched interface and boundary (MIB) method, we have recently developed a PB solver by rigorously enforcing the flux continuity conditions at the solvent-molecule interface where geometric singularities may occur. The resulting PB solver, denoted as MIBPB-II, is able to deliver second order accuracy for the molecular surfaces of proteins. However, when the mesh size approaches half of the van der Waals radius, the MIBPB-II cannot maintain its accuracy because the grid points that carry the interface information overlap with those that carry distributed singular charges. In the present Green's function formalism, the charge singularities are transformed into interface flux jump conditions, which are treated on an equal footing as the geometric singularities in our MIB framework. The resulting method, denoted as MIBPB-III, is able to provide highly accurate electrostatic potentials at a mesh as coarse as 1.2 A for proteins. Consequently, at a given level of accuracy, the MIBPB-III is about three times faster than the APBS, a recent multigrid PB solver. The MIBPB-III has been extensively validated by using analytically solvable problems, molecular surfaces of polyatomic systems, and 24 proteins. It provides reliable benchmark numerical solutions for the PB equation.


Arp2/3 complex is required for actin polymerization during platelet shape change.
Platelets undergo a series of actin-dependent morphologic changes when activated by thrombin receptor activating peptide (TRAP) or when spreading on glass. Polymerization of actin results in the sequential formation of filopodia, lamellipodia, and stress fibers, but the molecular mechanisms regulating this polymerization are unknown. The Arp2/3 complex nucleates actin polymerization in vitro and could perform this function inside cells as well. To test whether Arp2/3 regulated platelet actin polymerization, we used recombinant Arp2 protein (rArp2) to generate Arp2-specific antibodies (alpha Arp2). Intact and Fab fragments of alpha Arp2 inhibited TRAP-stimulated actin-polymerizing activity in platelet extracts as measured by the pyrene assay. Inhibition was reversed by the addition of rArp2 protein. To test the effect of Arp2/3 inhibition on the formation of specific actin structures, we designed a new method to permeabilize resting platelets while preserving their ability to adhere and to form filopodia and lamellipodia on exposure to glass. Inhibition of Arp2/3 froze platelets at the rounded, early stage of activation, before the formation of filopodia and lamellipodia. By morphometric analysis, the proportion of platelets in the rounded stage rose from 2.85% in untreated to 63% after treatment with alpha Arp2. This effect was also seen with Fab fragments and was reversed by the addition of rArp2 protein. By immunofluorescence of platelets at various stages of spreading, the Arp2/3 complex was found in filopodia and lamellipodia. These results suggest that activation of the Arp2/3 complex at the cortex by TRAP stimulation initiates an explosive polymerization of actin filaments that is required for all subsequent actin-dependent events.


The thrombotic potential of circulating tumor microemboli: computational modeling of circulating tumor cell-induced coagulation.
Thrombotic events can herald the diagnosis of cancer, preceding any cancer-related clinical symptoms. Patients with cancer are at a 4- to 7-fold increased risk of suffering from venous thromboembolism (VTE), with ∼7,000 patients with lung cancer presenting from VTEs. However, the physical biology underlying cancer-associated VTE remains poorly understood. Several lines of evidence suggest that the shedding of tissue factor (TF)-positive circulating tumor cells (CTCs) and microparticles from primary tumors may serve as a trigger for cancer-associated thrombosis. To investigate the potential direct and indirect roles of CTCs in VTE, we characterized thrombin generation by CTCs in an interactive numerical model coupling blood flow with advection-diffusion kinetics. Geometric measurements of CTCs isolated from the peripheral blood of a lung cancer patient prior to undergoing lobectomy formed the basis of the simulations. Singlet, doublet, and aggregate circulating tumor microemboli (CTM) were investigated in the model. Our numerical model demonstrated that CTM could potentiate occlusive events that drastically reduce blood flow and serve as a platform for the promotion of thrombin generation in flowing blood. These results provide a characterization of CTM dynamics in the vasculature and demonstrate an integrative framework combining clinical, biophysical, and mathematical approaches to enhance our understanding of CTCs and their potential direct and indirect roles in VTE formation.


The smoking estrogens - a potential synergy between estradiol and benzo(a)pyrene.
According to recent statistics, Lung Cancer (LC) is one of the most frequently diagnosed tumor types, representing nearly 12% of all global cancer cases. Moreover, in recent years, an increased mortality rate and incidence of this cancer were observed, especially among nonsmokers. Lung cancer patients are often characterized by poor prognosis and low survival rates, which encourages the scientific community to investigate the biochemical and molecular processes leading to the development of this malignancy. Furthermore, the mechanisms of LC formation and progression are not yet fully elucidated due to their high complexity, as well as a multitude of environmental, genetic, and molecular factors involved. Even though LC's association with exposure to cigarette smoke is indisputable, current research provides evidence that the development of this cancer can also be affected by the presence of estrogens and their interaction with several tobacco smoke components. Hence, the main goal of this brief review was to investigate reports of a possible synergy between 17β estradiol (E2), the most biologically active estrogen, and benzo(a)pyrene (BaP), a strongly carcinogenic compound produced as a result of incomplete tobacco combustion. The literature sources demonstrate a possible carcinogenic synergy between estrogens, especially E2, and BaP, a toxic tobacco smoke component. Therefeore, the combined effect of disturbed estrogen production in cancer cells, as well as the molecular influence exerted by BaP, could explain the increased aggressiveness and rate of LC development. Summarizing, the synergistic effect of these risk factors is an interesting area of further research.


High resolution IR spectroscopy of acetylene-furan in ultracold helium nanodroplets.
We have measured the IR spectrum of the acetylene-furan complex in ultracold helium nanodroplets in the region of the nu(3) CH(asym)-stretch vibration of the acetylene (between 3240 and 3300 cm(-1)). We have observed eight bands that can be attributed to acetylene-furan complexes. Two of these bands are assigned to two different isomers of the 1:1 acetylene-furan complex. The vibrational band at 3267.4 cm(-1) is assigned to the CH(asym)-stretch vibration of the dimer structure with the C-H of the acetylene being attached to the pi-system of the furan. The peak at 3272.1 cm(-1) is assigned to the CH(asym)-stretch vibration of the dimer structure with the C-H of the acetylene being attached to the oxygen atom of the furan. These assignments are confirmed by additional measurements of the spectrum of the (13)C-acetylene-furan complex.


Animal models of biliary injury and altered bile acid metabolism.
In the last 25years, a number of animal models, mainly rodents, have been generated with the goal to mimic cholestatic liver injuries and, thus, to provide in vivo tools to investigate the mechanisms of biliary repair and, eventually, to test the efficacy of innovative treatments. Despite fundamental limitations applying to these models, such as the distinct immune system and the different metabolism regulating liver homeostasis in rodents when compared to humans, multiple approaches, such as surgery (bile duct ligation), chemical-induced (3,5-diethoxycarbonyl-1,4-dihydrocollidine, DDC, α-naphthylisothiocyanate, ANIT), viral infections (Rhesus rotavirustype A, RRV-A), and genetic manipulation (Mdr2, Cftr, Pkd1, Pkd2, Prkcsh, Sec63, Pkhd1) have been developed. Overall, they have led to a range of liver phenotypes recapitulating the main features of biliary injury and altered bile acid metabolisms, such as ductular reaction, peribiliary inflammation and fibrosis, obstructive cholestasis and biliary dysgenesis. Although with a limited translability to the human setting, these mouse models have provided us with the ability to probe over time the fundamental mechanisms promoting cholestatic disease progression. Moreover, recent studies from genetically engineered mice have unveiled 'core' pathways that make the cholangiocyte a pivotal player in liver repair. In this review, we will highlight the main phenotypic features, the more interesting peculiarities and the different drawbacks of these mouse models. This article is part of a Special Issue entitled: Cholangiocytes in Health and Disease edited by Jesus Banales, Marco Marzioni, Nicholas LaRusso and Peter Jansen.


Testicular cancer and spermatogenesis.
We retrospectively analysed the characteristics of 54 men with testicular cancer. The group comprised 32 men with pure seminoma and 22 with non-seminoma germ cell tumours (NSGCT). This group was further compared to 190 healthy sperm donor candidates. Sperm quality was found to be higher in the seminoma versus the NSGCT patients in: sperm concentration [50 (0-230) versus 17 (0-288) x 10(6)/ml, P < 0.001], total motile sperm counts (TMC) [57 (0-508) versus 12 (0-854) x 10(6)/ejaculate, P = 0.002], post-thaw forward motile concentration [3 (0-28) versus 1.7 (0-17) x 10(6)/ml, P = 0.003] and motility percentage [20 (0-57) versus 12.5 (0-42) %, P = 0.002]. Serum hormone concentrations did not differ between these two sub-groups, although the follicle stimulating hormone concentrations were higher than normal in both (14.6 +/- 2.5 versus 10.4 +/- 1.4 mIU/ml, P > 0.05). As is well documented, cancer patients were found to have lower sperm quality compared to healthy candidates. The existence of these differences, and the fact that testicular cancer affects spermatogenesis, indicated that the mechanisms involved in the deterioration of sperm quality can, at least partially, be attributed to the type and origin of the malignant cancer. The higher sperm counts in the seminoma group may be related to the fact that the resemblance of the seminoma cells to normal germ cells is greater than that of the NSGCT cells, and therefore they retain a better capacity to function. Due to modern assisted reproductive technologies and micromanipulation achievements, the lower yield of spermatozoa in severe cases is no longer a major obstacle to offering cryopreservation to these patients.


Nobody Is Perfect: Comparison of the Accuracy of PCR-RFLP and KASP™ Method for Genotyping. ADH1B and FTO Polymorphisms as Examples.
DNA genotyping is among the most common analyses currently performed in scientific research. Two high-throughput genotyping techniques are widely used - the "classic" PCR-RFLP and probe-based methods such as TaqMan® PCR assay or KASP™ genotyping. The probe-based techniques are claimed to be more accurate than PCR-RFLP; however, the evidence for this claim is sparse. We have directly compared results of genotyping of two SNPs (rs1229984 and rs17817449) obtained by the PCR-RFLP and KASP™ in 1,502 adult Caucasians. The results were identical in 97.3 % and 95.9 % cases, respectively. Discrepancies (either different results or result obtained with one but not with the other method) were addressed by confirmatory analysis using direct sequencing. The sequencing revealed that both methods can give incorrect results, but the frequency of incorrect genotyping of rs1229984 and rs17817449 was very low for both methods - 0.1 % and 0.5 %, respectively, for PCR-RFLP and 0.1 % and 0.3 %, respectively, for KASP™. These results confirm that the KASP™ technique is slightly more accurate, but it achieves slightly lower call rates than PCR-RFLP. When carefully set up, both PCR-RFLP and KASP™ could have accuracy of 99.5 % or higher.


Polychlorinated dibenzo-p-dioxin and dibenzofuran concentrations in serum samples of workers at intermittently burning municipal waste incinerators in Japan.
To find whether or not incinerator workers employed at intermittently burning municipal incineration plants are exposed to high concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs). 20 Workers employed at three municipal waste incineration plants (incinerator workers) and 20 controls were studied. The previous job, dietary, smoking, and body weight and height were obtained from a questionnaire survey. Concentrations of PCDDs and PCDFs were measured in serum samples of the workers and the deposited dust of the plants. The influence of occupational exposure on concentrations of PCDDs and PCDFs in serum samples was examined by multiple regression analysis. Dust analysis showed that dominant constituents were octachlorodibenzo-p-dioxin (OCDD) and 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD) among the PCDDs, and 1,2,3,4,6,7,8-heptachlorodibenzofuran (HpCDF) and octachlorodibenzofuran (OCDF) among the PCDFs. The toxicity equivalents (TEQs) of summed PCDDs and PCDFs in the deposited dust were 0.91, 33, and 11 ng TEQ/g, respectively, for plants I, II, and III. The means of TEQ in serum samples of summed PCDDs and PCDFs in the incinerator workers and controls were 22.8 and 16.4 pg TEQ/g lipid for area I, 29.4 and 19.3 pg TEQ/g lipid for area II, and 22.8 and 24.9 pg TEQ/g lipid for area III, which were almost the same as for the general population of Japan. No significant differences in the TEQ of PCDDs and TEQ of PCDDs and PCDDs were found between the incinerator workers and the controls. However, the TEQ of PCDFs was significantly higher among the incinerator workers in areas I and II, and the 1,2,3,4,6,7,8-HpCDF concentration was also significantly higher for all three areas. When the occupational exposure index for each constituent of PCDDs and PCDFs was defined as the product of the duration of employment at the incineration plant and the concentration of the constituent in the deposited dust, multiple regression analysis showed that the concentrations of HxCDF, HpCDF, and TEQ of PCDFs in serum samples increased with the occupational exposure index. The multiple regression analysis also suggested that significant factors affecting the concentrations in serum samples were area for HxCDD, age for TCDD, PeCDD, PeCDF, TEQ of PCDDs, TEQ of PCDFs, and TEQ of summed PCDDs and PCDFs, and BMI for HxCDD, HpCDD, and OCDD. This study showed that incinerator workers employed at intermittently burning incineration plants were not necessarily exposed to high concentrations of PCDDs and PCDFs. However, the increases in the concentrations in serum of HxCDF, HpCDF and TEQ of PCDFs with the occupational exposure index suggest that the incinerator workers had inhaled dust containing PCDDs and PCDFs during their work.


[Inhibitory effect of Isorhamnetin and Hesperidin on LDL oxidation induced by Cu2+].
To evaluate the inhibitory effect of Isorhamnetin and Hesperidin on the LDL oxidation induced by Cu2+. The serum LDL was isolated by the one step density gradient ultracentrifugation. The LDL oxidation was induced by Cu2+ in vitro for different time periods. Isorhamnetin and Hesperidin at 5 micromol/L were added respectively, as the experimental groups, 3 hours before oxidation. The oxidation of LDL in experimental and control groups was identified by measuring A234 , REM, TBARS and protein carbonyls content. The values of A234, REM, TBARS and protein carbonyls formation increased gradually during LDL oxidation induced by Cu2+ in vitro. During LDL oxidation induced by Cu2+ in vitro and incubation with each of Isorhamnetin and Hesperidin, the kinetic changes of A234 , REM, TBARS and protein carbonyls formation showed lag phases of 2-4 h and 2 h respectively, and the corresponding values for each of the agents treated group were reduced by 23.5%-40.4%, 20.5%-37.7%, 18.6-30.3% and 20.1%-52.4% (P < 0.001); and 11.1%-21.2%, 9.2%-28.3%, 13.7%-21.3% and 5.0%-43.8% respectively (P < 0.001, P < 0.01, P < 0.001 and P < 0.05). It suggests that Isorhamnetin and Hesperidin can substantially inhibit LDL oxidation, and Isorhamnetin has antioxidation ability stronger than Hesperidin.


A protein component of Drosophila polar granules is encoded by vasa and has extensive sequence similarity to ATP-dependent helicases.
Determinants of pole cells, which are precursors of the germ line, are provided maternally and are localized to the posterior pole of the Drosophila egg, as are polar granules. It has been hypothesized that certain RNA molecules associated with polar granules may be necessary for pole cell determination. Using a monoclonal antibody (Mab46F11) against polar granules, we have cloned the gene for one of their components. This gene turns out to be vasa, which is required maternally for the formation of polar granules and germ cells. This polar granule component shows significant sequence similarity to eIF-4A, a translation initiation factor that binds to mRNA, and to other helicases.


Survey of antibodies against various infectious disease agents in racing camels in Abu Dhabi, United Arab Emirates.
Prevalence of antibodies against some important disease agents in sera from racing camels in Abu Dhabi (United Arab Emirates) is reported. Antibodies against Brucella abortus were detected in 1.5% of racing camels, but only 0.76% had titres sufficient for the animals to be considered infected. The complement fixation test revealed antibodies against Coxiella burnetii (causative agent of Q fever) in 7.9% of camels (with a geometrical mean titre of 13) and against parainfluenza virus type 3 in 5.6% of camels (with a geometrical mean titre of 20). Serological evidence did not show the presence of Aspergillus spp., while antibodies against Leptospira interrogans were seen in 4.1% of camels. Antibodies against Echinococcus polymorphus were observed in 2.6% of the animals, while a large number of racing camels (30.9% and 36.4%) possessed antibodies against Toxoplasma gondii, as determined by direct agglutination and indirect haemagglutination, respectively.


Salt fluoridation in France since 1986.
In 1985, the production and use of fluoridated salt was authorized in France. Domestic salt both with added fluoride (at 250 ppm) and without fluoride has been on the market since 1986. It was recommended to avoid the consumption of fluoridated salt if the local drinking water contained more than 0.5 milligrams of fluoride per litre. The legislation has never been modified since, except for a few developments such as the permission given to school canteens in 1993 to use fluoridated salt, provided canteen managers made sure that the drinking water contained no more than 0.5 milligrams of fluoride per litre. Epidemiological surveys about fluoridated salt in France are few, but they point in the same direction: decrease of DMFT and DMFS values, evident for the period 1986-1993 but minimal from 1993 to 1998. In 1999 and 2002, epidemiological comparative surveys were carried out in Montpellier (France) and Heidelberg (Germany) among 12-year-old schoolchildren. A decrease in caries prevalence was found in both towns, particulary in Heidelberg. On the other hand, the two cities showed some slight differences resulting from public health policy, from individual preventive habits, and from an earlier introduction of fluoridated salt in France.


[Changes of mortality and causes of death from 1970 to 2005 and decomposition analysis in Shandong province].
To describe the trend of overall mortality and major causes of death in Shandong population from 1970 to 2005, and to quantitatively estimate the influential factors. Trends of overall mortality and major causes of death were described using indicators such as mortality rates and age-adjusted death rates by comparing three large-scale mortality surveys in Shandong province. Difference decomposing method was applied to estimate the contribution of demographic and non-demographic factors for the change of mortality. The total mortality had had a slight change since 1970s, but had increased since 1990s. However, both the mortality rates of age-adjusted and age-specific decreased significantly. The mortality of Group I diseases including infectious diseases as well maternal and perinatal diseases decreased drastically. By contrast, the mortality of non-communicable chronic diseases (NCDs) including cardiovascular diseases (CVDs), cancer and injuries increased. The sustentation of recent overall mortality was caused by the interaction of demographic and non-demographic factors which worked oppositely. Non-demographic factors were responsible for the decrease of Group I disease and the increase of injuries. With respect to the increase of NCDs as a whole, demographic factors might take the full responsibility and the non-demographic factors were the opposite force to reduce the mortality. Nevertheless, for the increase of some leading NCD diseases as CVDs and cancer, the increase was mainly due to non-demographic rather than demographic factors. Through the interaction of the aggravation of ageing population and the enhancement of non-demographic effect, the overall mortality in Shandong would maintain a balance or slightly rise in the coming years. Group I diseases in Shandong had been effectively under control. Strategies focusing on disease control and prevention should be transferred to chronic diseases, especially leading NCDs, such as CVDs and cancer.


Advantages and limitations of density gradient ultracentrifugation in the fractionation of human serum lipoproteins: role of salts and sucrose.
Two density gradient ultracentrifugation methods, Redgrave et al. (1975. Anal. Biochem. 65: 42-49) and Nilsson et al. (1981. Anal. Biochem. 110: 342-348), currently used for the separation and analysis of plasma lipoproteins were compared with respect to their resolving power and capacity to obtain pure products as a function of time of ultracentrifugation using the same rotor (Beckman SW-40), speed (150,000 g), and temperature (14 degrees C). The effects of sucrose and salts were also investigated. The Redgrave gradient insured the separation of the major classes of plasma lipoproteins after 24 hr of centrifugation; however, equilibrium conditions were only reached after 48 hr, at which time the lipoproteins were contaminated by albumin. When the effluents from each rotor tube were continuously monitored at 280 nm, each lipoprotein band gave values that were higher than those from mass analyses. This was due to a light scattering effect, the extent of which was dependent on the concentration of lipoproteins and salts. Sucrose prevented the scattering effect and was found to bind irreversibly to the apolipoproteins. In contrast, after 66 hr centrifugation, the lipoproteins obtained from the Nilsson gradient exhibited a close correspondence between protein mass and absorbance values at 280 nm, had no scattering effect, and were uncontaminated by albumin. The difference in spectroscopic behavior between the Redgrave and the Nilsson procedures was attributed to three factors: 1) the presence of sucrose in the latter gradient and incorporation of this sugar into lipoproteins as assessed by mass and radioactivity measurements; 2), the salt density to which the serum samples were exposed to at the beginning of the ultracentrifugation; and 3) the final lipoprotein concentration.(ABSTRACT TRUNCATED AT 250 WORDS)


[Progress in the research of Chinese medical history in recent five years].
In recent five years, the research of Chinese medical history has made new progress on the basis of what has already been achieved. National conferences on history of Chinese medicine, including minority nationalities, modern and contemporary, medical exchanges with foreign countries and its comparative study on medical history have been convened. After those, research on those domains has made marked progress. The relevant theses and monographs has increased distinctly. A general history of Chinese medicine, organized by the Ministry of Public Health, will propel the research on general history of medicine profoundly. In addition, research on history of medical subjects and diseases, medical personages and works, theory of Chinese medical history, history of foreign medicine have also made new progress. There are frequent academic exchanges internationally day by day. But, there are also questions that exist; the drain of talented people, the historical course of Chinese medicine has not been included in the lust of continuous education in the Chinese Medical Association, weak points in research on history of diseases and foreign medicine, the historical theory of Chinese medicine and so on. These questions remain to be solved and strengthened. New domain on the medical history in Hongkong, Macao, and Taiwan should also be speeded up.


Effects of Oral Commensal Streptococci on Porphyromonas gingivalis Invasion into Oral Epithelial Cells.
The objective of this study was to determine if the interaction between common oral commensal bacteria and oral epithelial cells would provide protective effects against the invasion of periodontopathogen Porphyromonas gingivalis. Oral epithelial OKF6/Tert cells were used in co-cultures with Streptococcus gordonii, Streptococcus oralis, Streptococcus mitis, and Streptococcus intermedius. The viability of OKF6/Tert cells following a bacterial challenge was evaluated by trypan blue exclusion. The adherence of commensal species was determined by CFU counts. P. gingivalis invasion in OKF6/Tert cells was assessed before and after exposure to commensal species according to CFU counts. Viability assays show that only S. gordonii and S. intermedius display low toxicity toward OKF6/Tert cells. Both commensals adhere to OKF6/Tert cells at an average ratio of 1 CFU to 10 cells. P. gingivalis invasion into host cells is significantly reduced by 25% or 60% after exposure to S. gordonii or S. intermedius, respectively. The results suggest that these commensal species bind to host cells and diminish P. gingivalis invasion. This is important in the context of periodontal disease since P. gingivalis primarily acts on the host by invading it. Therefore, efforts to decrease invasion will eventually lead to future therapies harnessing the mechanisms employed by oral commensal bacteria.


Highly Efficient Organic Blue Electroluminescent Materials and Devices with Mesoscopic Structures.
Due to the difficulty in achieving high efficiency and high color purity simultaneously, blue emission is the limiting factor for the performance and stability of OLEDs. Since 2003, we have been working on organic light-emitting diodes (OLEDs), especially on blue light. After a series of molecular designs, novel strategies have been proposed from different aspects. At first, highly efficient deep blue emission could be achieved through molecular design with highly twisted structure to suppress fluorescence quenching and redshift. Deep blue emitters with high efficiency in solid state, a twisted structure with aggregation induced emission (AIE) characteristics was incorporated to inhibit molecular aggregation, and triplet-triplet fusion (TTF) and hybridized localized charge transfer (HLCT) were adopted to increase the ratio of triplet exciton used. Secondly, a highly efficient blue OLED could be achieved through improving charge transport. New electron transport materials (ETMs) with wide band gap were developed to control charge transport balance in devices. Thirdly, a highly efficient deep blue emission could be achieved through a mesoscopic structure of out-coupling layer. A mesoscopic photonic structured organic thin film was fabricated on the top of metal electrode by self-aggregation in order to improve the light out-coupling efficiency.


Lens transglutaminase selects specific beta-crystallin sequences as substrate.
A Ca2+-dependent transglutaminase (EC 188.8.131.52) has been demonstrated in the eye lenses of several mammalian species [Lorand, L., Hsu, L. K. M., Siefring, G. E., Jr., & Rafferty, N. S. (1981) Proc. Natl. Acad. Sci. USA 78, 1356-1360]. Using [3H]methylamine as a convenient probe for transglutaminase activity, we have explored the action of this enzyme in the bovine eye lens. We could characterize the glutamine residues acting as acyl-donor sites in three beta-crystallin chains, which are the only substrates for lens transglutaminase among the various lens-specific structural proteins, the crystallins. A single glutamine was found to bind [3H]methylamine in each of these three chains: glutamine -9 in beta Bp (beta B2), glutamine -21 in beta B3, and glutamine -23 or -24 in beta A3. The four glutamines are all located in the NH2-terminal regions, which presumably extend from the compact two-domain structure of the beta-crystallin chains. It was, moreover, established that several components of the lens cytoskeleton are substrates for transglutaminase.


Transcription factor interplay in T helper cell differentiation.
The differentiation of CD4 helper T cells into specialized effector lineages has provided a powerful model for understanding immune cell differentiation. Distinct lineages have been defined by differential expression of signature cytokines and the lineage-specifying transcription factors necessary and sufficient for their production. The traditional paradigm of differentiation towards Th1 and Th2 subtypes driven by T-bet and GATA3, respectively, has been extended to incorporate additional T cell lineages and transcriptional regulators. Technological advances have expanded our view of these lineage-specifying transcription factors to the whole genome and revealed unexpected interplay between them. From these data, it is becoming clear that lineage specification is more complex and plastic than previous models might have suggested. Here, we present an overview of the different forms of transcription factor interplay that have been identified and how T cell phenotypes arise as a product of this interplay within complex regulatory networks. We also suggest experimental strategies that will provide further insight into the mechanisms that underlie T cell lineage specification and plasticity.


Fetal testosterone and sex differences in typical social development and in autism.
Experiments in animals leave no doubt that androgens, including testosterone, produced by the testes in fetal and/or neonatal life act on the brain to induce sex differences in neural structure and function. In human beings, there is evidence supporting a female superiority in the ability to read nonverbal signals, specific language-related skills, and theory of mind. Even more striking than the sex differences seen in the typical population is the elevated occurrence of social and communicative difficulties in human males. One such condition, autism, occurs four times more frequently in boys than in girls. Recently, a novel theory known as the "extreme male brain" has been proposed. It suggests that the behaviors seen in autism are an exaggeration of typical sex differences and that exposure to high levels of prenatal testosterone might be a risk factor. In this article, we argue that prenatal and neonatal testosterone exposures are strong candidates for having a causal role in sexual dimorphism in human behavior, including social development, and as risk factors for conditions characterized by social impairments, particularly autism spectrum conditions.


Transcriptome programs involved in the development and structure of the cerebellum.
In the past two decades, mounting evidence has modified the classical view of the cerebellum as a brain region specifically involved in the modulation of motor functions. Indeed, clinical studies and engineered mouse models have highlighted cerebellar circuits implicated in cognitive functions and behavior. Furthermore, it is now clear that insults occurring in specific time windows of cerebellar development can affect cognitive performance later in life and are associated with neurological syndromes, such as Autism Spectrum Disorder. Despite its almost homogenous cytoarchitecture, how cerebellar circuits form and function is not completely elucidated yet. Notably, the apparently simple neuronal organization of the cerebellum, in which Purkinje cells represent the only output, hides an elevated functional diversity even within the same neuronal population. Such complexity is the result of the integration of intrinsic morphogenetic programs and extracellular cues from the surrounding environment, which impact on the regulation of the transcriptome of cerebellar neurons. In this review, we briefly summarize key features of the development and structure of the cerebellum before focusing on the pathways involved in the acquisition of the cerebellar neuron identity. We focus on gene expression and mRNA processing programs, including mRNA methylation, trafficking and splicing, that are set in motion during cerebellar development and participate to its physiology. These programs are likely to add new layers of complexity and versatility that are fundamental for the adaptability of cerebellar neurons.


Drug addiction. Part II. Neurobiology of addiction.
The drug addiction may be regarded as the disease of the brain reward system. This system, closely related to the system of emotional arousal, is located predominantly in the limbic structures of the brain. Its existence was proved by demonstration of the "pleasure centers," that were discovered as location from which electrical self-stimulation is readily evoked. The main neurotransmitter involved in the reward is dopamine, but other monoamines and acetylcholine may also participate. The anatomical core of the reward system are dopaminergic neurons of the ventral tegmentum that project to the nucleus accumbens, amygdala, prefrontal cortex and other forebrain structures. Several of those structures may be specifically involved in the reward produced by different substances, when anticipating the reward. The recent discovery of CART peptides may importantly expand our knowledge about the neurochemistry of reward. Natural rewarding activities and artificial chemical rewarding stimuli act at the same locations, but while natural activities are controlled by feedback mechanisms that activate aversive centers, no such restrictions bind the responses to artificial stimuli. There are several groups of substances that activate the reward system and they may produce addiction, which in humans is a chronic, recurrent disease, characterized by absolute dominance of drug-seeking behavior. The craving induced by substances of addiction inhibits other behaviors. The adaptation of an organism to a chronic intake of drugs involves development of adaptive changes, sensitization or tolerance. It is thought that the gap between sensitization developing for the incentive value of the drug and tolerance to the reward induced by its consumption underlies the vicious circle of events leading to drug dependence. The vulnerability to addiction is dependent not only on the environment, but also on genetic factors.


The microscopic anatomy of epidermal glands in two species of gekkonine lizards, with some observations on testicular activity.
The gross and microscopic anatomy of epidermal glands has been studied in laboratory maintained tokays (Gekko gecko), and house geckos (Hemidactylus bowringii) captured from the wild throughout the year. Annual testicular activity in the house gecko has also been studied. While no significant differences in glandular development at various times have been observed in G. gecko, there are clear-cut annual cycles in H. bowringii. The evolution of epidermal glands in gekkonid lizards is reviewed; the cellular dynamics of beta-glands are compared with those of unspecialized epidermis; the possibility that gekkonine epidermal glands respond to quantitative variation in circulating testosterone titers is discussed.


Anticardiolipin antibodies and giant cell arteritis: a prospective, multicenter case-control study. Groupe de Recherche sur l'Artérite à Cellules Géantes.
To assess the prevalence and thrombogenic role of anticardiolipin antibodies (aCL) in giant cell arteritis. Two hundred eighty-four patients with newly diagnosed temporal arteritis or polymyalgia rheumatica and 210 age- and sex-matched controls randomly selected from the general population were included in a multicenter, prospective case-control study. Blood samples were obtained at the time of diagnosis, data on initial clinical features were collected in a questionnaire, and temporal artery biopsy findings were reviewed by an experienced pathologist. Anticardiolipin antibodies were present in 20.7% of patients compared with 2.9% of controls (P = 1.45 x 10(-9)). The prevalence of aCL was higher in samples found positive for temporal arteritis than in those found negative on biopsy (31.2% versus 16.7%; P = 0.04), and was similarly higher in the biopsy-positive temporal arteritis group compared with the polymyalgia rheumatica and control groups. Although aCL were associated with thrombotic complications in univariate analysis, the positivity of the biopsy findings remained the only predictive variable in stratified analysis. In giant cell arteritis, aCL seem to function as reactive antibodies in relation to endothelial lesions.


Hemoglobin variants in Koreans: hemoglobin G Taegu.
Hemoglobin G Taegu, an electrophoretically slow hemoglobin with a structural anomaly believed to be in the beta-T-3 section of the beta chain, was the only variant found among 6700 normal Koreans. Four subjects, 0.06 percent, had the G-hemoglobin variant in addition to normal hemoglobin A. Hemoglobin E, known in numerous groups from Southeast Asia and the variant most frequently seen in Chinese subjects, was not found among the Koreans we tested.


Reactive Oxygen Species in Cardiovascular Calcification: Role of Medicinal Plants.
Cardiovascular calcification, including vascular calcification and calcific aortic valve disease (CAVD), is a serious worldwide health problem, especially in older adults. The mechanisms underlying cardiovascular calcifications are complex and multifactorial. An increase in reactive oxygen species (ROS) and oxidative stress play important roles in the initiation and development of cardiovascular calcification. This mini-review summarizes the recent evidence that supports the association of ROS with vascular calcification and CAVD and discusses the role of medicinal plants for the prevention and treatment of cardiovascular calcification.


Uptake of an Incentive-Based mHealth App: Process Evaluation of the Carrot Rewards App.
Behavioral economics has stimulated renewed interest in financial health incentives worldwide. The Carrot Rewards app was developed as part of a public-private partnership to reward Canadians with loyalty points (eg, movies and groceries) for downloading the app, referring friends, and completing an average of 1 to 2 educational health quizzes per week ("micro-learning"), with long-term objectives of increasing health knowledge and encouraging healthy behaviors. The main objective of this study was to evaluate uptake of a loyalty points-based mHealth app during the exclusive 3-month launch period in British Columbia (BC), Canada. The secondary aims were to describe the health and sociodemographic characteristics of users, as well as participation levels (eg, proportion of quizzes completed and friends referred). The app was promoted via loyalty program email campaigns (1.64 million emails). Number of downloads and registrations (users enter age, gender, and valid BC postal code to register) were collected. Additional sociodemographics were inferred by linking postal codes with census data at the local health area (LHA) level. Health risk assessments were also deployed. Participation levels were collected over 3 months and descriptive data were presented. In 3 months, 67,464 individuals downloaded the app; in its first week, Carrot Rewards was the most downloaded health app in Canada. Among valid users (n=57,885; at least one quiz completed), the majority were female (62.96%; 36,446/57,885) and aged 18 to 34 years (54.34%; 31,459/57,885). More than half of the users (52.40%; 30,332/57,885) resided in LHAs where the median personal income was below the provincial average (Can $28,765). Furthermore, 64.42% (37,291/57,885) of users lived in metropolitan (ie, urban) LHAs, compared with 56.17% of the general BC population. The most prevalent risk factors were "not" meeting physical activity guidelines (72.70%; 31,765/43,692) and "not" getting the flu shot last year (67.69%; 30,286/44,739). Regarding participation, 60.05% (34,761/57,885) of users were classified as "very high" engagers (>75% quiz completion rate). Early results suggest that loyalty points may promote mHealth app uptake. The app was downloaded by younger females especially, and BC residents from higher and lower income regions were equally represented. Loyalty points appear to have driven participation throughout the inaugural 3-month period (ie, quiz completion).


Time-Resolved Fluorescence Anisotropy of a Molecular Rotor Resolves Microscopic Viscosity Parameters in Complex Environments.
Understanding viscosity in complex environments remains a largely unanswered question despite its importance in determining reaction rates in vivo. Here, time-resolved fluorescence anisotropy imaging (TR-FAIM) is combined with fluorescent molecular rotors (FMRs) to simultaneously determine two non-equivalent viscosity-related parameters in complex heterogeneous environments. The parameters, FMR rotational correlation time and lifetime, are extracted from fluorescence anisotropy decays, which in heterogeneous environments show dip-and-rise behavior due to multiple dye populations. Decays of this kind are found both in artificially constructed adiposomes and in live cell lipid droplet organelles. Molecular dynamics simulations are used to assign each population to nano-environments within the lipid systems. The less viscous population corresponds to the state showing an average 25° tilt to the lipid membrane normal, and the more viscous population to the state showing an average 55° tilt. This combined experimental and simulation approach enables a comprehensive description of the FMR probe behavior within viscous nano-environments in complex, biological systems.


The Effectiveness of Varroa destructor Infestation Classification Using an E-Nose Depending on the Time of Day.
Honey bees are subject to a number of stressors. In recent years, there has been a worldwide decline in the population of these insects. Losses raise a serious concern, because bees have an indispensable role in the food supply of humankind. This work is focused on the method of assessment of honey bee colony infestation by Varroa destructor. The approach allows to detect several categories of infestation: "Low", "Medium" and "High". The method of detection consists of two components: (1) the measurements of beehive air using a gas sensor array and (2) classification, which is based on the measurement data. In this work, we indicate the sensitivity of the bee colony infestation assessment to the timing of measurement data collection. It was observed that the semiconductor gas sensor responses to the atmosphere of a defined beehive, collected during 24 h, displayed temporal variation. We demonstrated that the success rate of the bee colony infestation assessment also altered depending on the time of day when the gas sensor array measurement was done. Moreover, it was found that different times of day were the most favorable to detect the particular infestation category. This result could indicate that the representation of the disease in the beehive air may be confounded during the day, due to some interferences. More studies are needed to explain this fact and determine the best measurement periods. The problem addressed in this work is very important for scheduling the beekeeping practices aimed at Varroa destructor infestation assessment, using the proposed method.


Genetic influences on joint contractures secondary to immobilization.
The primary research question of this study queries whether, beyond environmental conditions, genetic factors affect the development of joint contractures. We hypothesized that intrinsic genetic factors influence the severity of joint contractures developing secondary to joint immobilization. Forty rats from four inbred rat strains had one leg immobilized in knee flexion for 4 weeks. The contracture was measured mechanically as the lack of range of motion to a standardized torque. Using the contralateral leg as a control, the average severity of the contracture could be calculated and compared between strains. All immobilized legs presented knee contractures after 4 weeks of immobilization. Two strains (Dark Agouti and Fisher 344) showed a larger mean knee contracture than those of the two other rat strains (Augustus Copenhagen Irish and Brown Norway). Environmental factors, such as immobility, are usually identified as a cause of a joint contracture. These results demonstrate that, in addition to mechanical factors in the environment of a joint, intrinsic genetic factors participate in the process leading to joint contracture. This demonstration has important consequences for directing future research and may lead to interventions to help patients at risk of developing joint contractures.


Rectangular lattice Boltzmann method.
A set of rectangular lattice Boltzmann methods for fluid flows is developed. It is shown that reformulating local equilibrium distribution functions can result in the rectangular lattice Boltzmann models without the aid of an interpolation for shallow water equations, Navier-Stokes equations, and axisymmetric flow equations. In addition, schemes for correct incorporation of force terms into the models are proposed for simulations of flows involving forces in practice. The methods completely retain the innate kinetic features and the simple procedure of the standard lattice Boltzmann method with an additional benefit of being suitable for rectangular lattices at little extra computational cost. The methodology is illustrated and validated through its application to different flow problems, demonstrating the potential power of the models.


The associations of alcoholic liver disease and nonalcoholic fatty liver disease with bone mineral density and the mediation of serum 25-Hydroxyvitamin D: A bidirectional and two-step Mendelian randomization.
Reduced bone mineral density (BMD) and osteoporosis are common in chronic liver diseases. However, the causal effect of alcoholic liver disease (ALD) and non-alcoholic fatty liver disease (NAFLD) on BMD remains uncertain. This study uses a two-sample Mendelian randomization (MR) design to evaluate the genetically predicted effect of ALD and NAFLD on BMDs using summary data from publically available genome-wide association studies (GWASs). The GWAS summary statistics of ALD (1416 cases and 213,592 controls) and NAFLD (894 cases and 217,898 controls) were obtained from the FinnGen consortium. BMDs of four sites (total body, n = 56,284; femoral neck, n = 32,735; lumbar spine, n = 28,498; forearm, n = 8143) were from the GEnetic Factors for OSteoporosis Consortium. Data for alcohol consumption (n = 112,117) and smoking (n = 33,299) and serum 25-Hydroxyvitamin D (25-OHD) level (n = 417,580) were from UK-biobank. We first performed univariate MR analysis with the Inverse Variance Weighted (IVW) method as the primary analysis to investigate the genetically predicted effect of ALD or NAFLD on BMD. Then, multivariate MR and mediation analysis were performed to identify whether the effect was mediated by alcohol consumption, smoking, or serum 25-OHD level. The MR results suggested a robust genetically predicted effect of ALD on reduced BMD in the femoral neck (FN-BMD) (IVW beta = -0.0288; 95% CI: -0.0488, -0.00871; P = 0.00494) but not the other three sites. Serum 25-OHD level exhibited a significant mediating effect on the association between ALD and reduced FN-BMD albeit the proportion of mediation was mild (2.21%). No significant effects of NAFLD, alcohol consumption, or smoking on BMD in four sites, or reverse effect of BMD on ALD or NAFLD were detected. Our findings confirm the genetically predicted effect of ALD on reduced FN-BMD, and highlight the importance of periodic BMD and serum 25-OHD monitoring and vitamin D supplementation as needed in patients with ALD. Future research is required to validate our results and investigate the probable underlying mechanisms.


Plant bioactives for ruminant health and productivity.
Plants have been used throughout history for their medicinal properties. This use has often focused on human health but plants have also been, and still are, applied in ethnoveterinary practice and animal health management. In recent times, the use of synthetic chemicals has become prevalent. Public awareness of the potential environmental and health risks associated with heavy chemical use has also increased. This has put pressure on regulatory bodies to reduce the use of chemicals in agriculture. The most striking example is the 2006 banning of antibiotics in animal feed by the European Union. Moves such as this have increased the drive to find alternatives to synthetic chemicals and research has again turned to the use of plant bioactives as a means of improving animal health. Current scientific evidence suggests there is significant potential to use plants to enhance animal health in general and that of ruminants (cattle, deer, sheep, etc.) in particular. Active areas of research for plant bioactives (particularly saponin and tannin containing plants) include reproductive efficiency, milk and meat quality improvement, foam production/bloat control and methane production. Nematode control is also a significant area of research and the evidence suggests a much broader range of phytochemicals may be effective. This review presents a summary of the literature and examines international research efforts towards the development of plant bioactives for animal health.


Structurally similar TCRs differ in their efficiency of positive selection.
Studies of TCR transgenic mice have demonstrated that in these systems positive selection is not an efficient process. The capacity of the thymus to produce mature T cells is limited, even when all immature thymocytes express appropriate Ag receptors. Analysis of TCR transgenic mice expressing reduced levels of MHC molecules have shown that MHC surface density can be a limiting factor during development. Whether peptide availability in the thymus also limits the efficiency of positive selection remains controversial. Here, we examine the efficiency of positive selection in three similar lines of TCR transgenic mice, all of which express V alpha11/beta3+ TCRs specific for cytochrome c peptides bound to I-Ek. We demonstrate that thymocytes expressing these similar TCRs mature with very different efficiencies in H-2k mice. Furthermore, efficient positive selection of thymocytes expressing these three TCRs varies in its dependence on MHC density. These data suggest that similar TCRs can differ in their degree of specificity for peptide/MHC complexes during positive selection; some TCRs may require specific complexes for selection while other TCRs are more promiscuous in their thymic interactions. Alternatively, these three TCRs may differ in their affinities for positively selecting ligands.


Instruments for the assessment of suicide risk: A systematic review evaluating the certainty of the evidence.
Instruments have been developed to facilitate suicide risk assessment. We aimed to evaluate the evidence for these instruments including assessment of risk of bias and diagnostic accuracy for suicide and suicide attempt. PubMed (NLM), PsycInfo, Embase, Cinahl and the Cochrane Library databases were searched until December 2014. We assessed risk of bias with QUADAS-2. The average sensitivity and specificity of each instrument was estimated and the certainty of the evidence was assessed with GRADE. We considered instruments with a sensitivity > 80% and a specificity > 50% to have sufficient diagnostic accuracy. Thirty-five relevant studies were identified but 14 were considered to have high risk of bias, leaving 21 studies evaluating altogether 15 risk assessment instruments. We could carry out meta-analyses for five instruments. For the outcome suicide attempt SAD PERSONS Scale had a sensitivity of 15% (95% CI 8-24) and specificity of 97% (96-98), and the Manchester Self-Harm Rule (MSHR) a sensitivity of 97% (97-97) and a specificity of 20% (20-21). ReACT, which is a modification of MSHR, had a similar low specificity, as did the Sodersjukhuset Self Harm Rule. For the outcome suicide, the Beck Hopelessness Scale had a sensitivity of 89% (78-95) and specificity of 42% (40-43). Most suicide risk assessment instruments were supported by too few studies to allow for evaluation of accuracy. Among those that could be evaluated, none fulfilled requirements for sufficient diagnostic accuracy.


Seasonal variations, metal distribution and water quality in the Todos os Santos River, Southeastern Brazil: a multivariate analysis.
In aquatic habitats, metal contamination from natural and anthropogenic sources continues to pose a concern for human and environmental health. Thus, it is important to complete monitoring studies to assess patterns and the extent of metal contamination in these ecosystems. The purpose of this work was to determine the concentrations of 31 chemical elements and water quality parameters of the Todos os Santos River located in the Mucuri Valley, Minas Gerais, Brazil. A multivariate statistical analysis was used to determine any seasonal and spatial patterns from the data. Results demonstrated that metals including Al, Fe, and Ni exceeded Brazilian and international guidelines nutrients as P also exceed water quality standards. Principal components analysis indicated distinct geographical and seasonal patterns for multiple elements with hierarchical cluster analysis confirming the observed spatial patterns of contamination in the Todos os Santos River.


Comparison between the effect of 6 weeks of morning or evening aerobic exercise on appetite and anthropometric indices: a randomized controlled trial.
Several studies have shown that exercise is directly related to creating negative energy balance and changes in appetite. However, few studies have examined the effect of exercise time during the day on these factors. The aim of the present study was to demonstrate the effect of 6 weeks of morning and evening aerobic exercise on appetite and anthropometric indices. A total of 48 overweight females were recruited to this clinical trial. By the time of exercise, they were divided into two groups (morning or evening) and performed 6 weeks of exercise with a target heart rate on the ventilatory threshold. Appetite change, calorie intake and anthropometric indices were assessed. Consistent changes in appetite scores were not found during the 6 weeks (P > 0.05). Calorie consumption of the morning group decreased significantly more than that of the evening group (P = 0.02) during the 6 weeks. In addition, significant changes in body weight, body mass index, abdominal skin fold thickness and abdominal circumference were seen in the morning group. It appears that moderate- to high-intensity aerobic exercise in the morning could be considered a more effective programme than evening exercise on appetite control, calorie intake and weight loss in inactive overweight women. However, the limitations of the study, such as short-term duration, should be noticed.


Enhancer-like stimulation of yeast tRNA gene expression by a defined region of the Ty element micro-injected into Xenopus oocytes.
In the yeast Saccharomyces cerevisiae, the majority of the tRNA genes are found associated with transposable elements one of which is the Ty element. We noticed that the transcriptional activity of several of these genes was rather different depending on the type of the accompanying element(s) [Nelböck et al. (1985) Biol. Chem. Hoppe-Seyler 366, 1041-1051]. In order to study the influence of a Ty element on tRNA gene expression, we used the method of micro-injection into Xenopus oocytes taking advantage of a Ty+/Ty- allelic pair of a tRNALys1 gene recently isolated in our laboratory. A direct comparison showed that the expression of the plus allele was c. sixfold higher than that of the minus allele. In order to determine sequences of the Ty responsible for this effect, we constructed a number of variants in which distinct segments of the Ty were deleted or rearranged. The activating ability was localized to a 590-bp segment of the Ty containing the 'promoter delta', independent of its orientation or location towards the tRNA gene thus resembling the effects described for transcriptional enhancers. This is the first time that a long-range effect has been observed on the expression of a gene transcribed by RNA polymerase III.


Classification and technology of byzantine mosaic glass.
Glass-making is a very sophisticated skill and the contribution given by the chemical analyses of glass materials is fundamental for the classification of glass types and for identifying compositional groups according to consistent characteristics that can be associated with chronological and geographical differentiations. The chemical composition of glasses is particularly complex: to a few basic constituents many components were added, either derived from impurities in the raw materials or intentionally incorporated into the glass mix. The field of study concerning the chemical composition and the technology of Byzantine mosaic production has not been dealt with in a systematic manner and certainly not exhaustively from the view point of classification according to the reconstruction of chronological and geographical development. Nevertheless, it is of great interest because it is probable that during the Byzantine period the production of mosaic glass was greater than for any other type of glass. We propose a methodology for classifying Byzantine mosaic glasses on the basis of simple statistical treatment of the chemical composition data. Compositional data relative to basic and accessory constituents together with colorants were analysed and elaborated through binary diagrams. Byzantine glasses are also compared to glasses of different epoch and provenance.


Mass spectrometry of prostaglandins, leukotrienes and steroids as their allyldimethylsilyl ether derivatives.
Allyldimethylsilyl ethers of prostaglandin E2 and F2 alpha, leukotriene B4 and 5 beta-pregnane-3 alpha, 20 alpha-diol were prepared at room temperature in good yields with the novel reagent N,O-bis(allyldimethylsilyl)-trifluoroacetamide. The gas chromatographic properties of the derivatives of prostaglandins and the steroid were found to be excellent whereas those of leukotriene B4 were found to be less than satisfactory. The mass spectra of the allyldimethylsilyl ether derivatives of the compounds studied show intense ions in their upper mass region derived from the elimination of an allyl radical. In the mass spectrum of the derivative of leukotriene B4, the formation of a conjugated carbonium ion by alpha-cleavage is a major process. The detection limits for a quantitative assay were established by performing selected ion monitoring on the most intense ion in the upper mass region of respective mass spectrum. Detection limits in the low picogram range were obtained for the prostaglandins and the steroid but the allyldimethylsilyl ether derivative of leukotriene B4 exhibited a far higher detection limit. It is concluded for the latter compound that the detection limit depends primarily on the gas chromatographic properties.


Effect of biochar addition on hydrogen and methane production in two-phase anaerobic digestion of aqueous carbohydrates food waste.
Effect of biochar addition on hydrogen and methane production in two-phase anaerobic digestion of aqueous carbohydrates was studied using bench-scale bioreactors. The cultures with biochar additions were placed in 100ml reactors and incubated at 35°C and pH 5 for hydrogen production. The residual cultures were then used for methane production, incubated at 35°C and pH 7. Daily yields of hydrogen and methane and weekly yield of volatile fatty acids (VFA) were measured. The hydrogen and methane production potentials, rate and lag phases of the two phases were analysed using the Gompertz model. The results showed that biochar addition increased the maximum production rates of hydrogen by 32.5% and methane 41.6%, improved hydrogen yield by 31.0% and methane 10.0%, and shortened the lag phases in the two phases by 36.0% and 41.0%, respectively. Biochar addition also enhanced VFA generation during hydrogen production and VFA degradation in methane production.


Developing a reflective standard of care.
Developing and monitoring effective practice is essential in health care organisations. Reflective practice offers practitioners and organisations a means to understand and learn through experience in complex care situations and develop expertise. This paper describes standard-setting in a hospice to construct a standard for managing patients who exhibit aggressive behaviour as part of their terminal illness.


The influence of cross unconditional stimulus reinstatement on electrodermal responding and conditional stimulus valence in differential fear conditioning.
We examined whether the inhibitory Conditional Stimulus (CS)-no Unconditional Stimulus (US) association formed during extinction can be triggered by a novel US during the reinstatement of conditional electrodermal responding and self-reported CS valence in human differential fear conditioning. Participants were trained with either a shock or an aversive scream US before undergoing extinction. Participants then received either the same (i.e., shock_shock or scream_scream) or a different US during reinstatement (i.e., shock_scream, scream_shock). Differential conditioning across all indices was stronger when a shock US was used during acquisition. After reinstatement, electrodermal responding to both the CS+ and the CS- increased regardless of the type of US used during reinstatement (non-differential reinstatement). Differential CS valence evaluations were larger after reinstatement in the groups that received the same US during acquisition and reinstatement (differential reinstatement), but differential evaluations did not increase in the groups receiving a different US at reinstatement. This dissociation suggests that the reinstatement of negative stimulus valence and the reinstatement of expectancy learning may differ.


The effect of the COVID-19 pandemic on routine childhood immunization coverage and timeliness in India: retrospective analysis of the National Family Health Survey of 2019-2021 data.
The COVID-19 pandemic has disrupted health systems globally. We estimated the effect of the pandemic on the coverage and timeliness of routine childhood immunization in India through April 2021. We used data from India's National Family Health Survey 2019-2021 (NFHS-5), a cross-sectional survey which collected immunization information of under-five children from a nationally representative sample of households between June 2019 and April 2021. We used a mother fixed-effects regression model - accounting for secular trends and confounding factors - to compare COVID-affected children with their COVID-unaffected siblings (n = 59,144). Children who were eligible for a vaccine after January 30, 2020 (date of the first COVID case in India) were considered as the COVID-affected group and those eligible for a vaccine before this date were included in the COVID-unaffected group. Coverage of the following vaccine doses was considered-Bacillus Calmette-Guérin (BCG), hepatitis B birth dose (hepB0), DPT1 (diphtheria, pertussis, and tetanus, first dose), DPT2, DPT3, polio1, polio2, polio3, and measles first dose (MCV1). Indicators of vaccine coverage and vaccine timeliness (defined as receiving a dose within 45 days of minimum eligibility age) were separately examined. Immunization coverage was lower in COVID-affected children as compared with unaffected children, ranging from 2% lower for BCG and hepB0 to 9% for DPT3 and 10% for polio3. There was no significant difference in MCV1 coverage. Coverage reduction was greater for vaccines doses given in later age groups. The rate of timely receipt of polio and DPT vaccine doses was 3%-5% lower among COVID-affected children relative to unaffected children. Among population subgroups, COVID-affected male children and those from rural areas experienced the highest reduction in vaccine coverage. Children in India experienced lower routine immunization coverage and greater delays in immunization during the COVID-19 pandemic. The Bill & Melinda Gates Foundation.


Diabetes and obesity during pregnancy are associated with oxidative stress genotoxicity in newborns.
Objective To compare the level of oxidative deoxyribonucleic acid (DNA) damage (genotoxicity) between the offspring of mothers with and without diabetes diagnosed during pregnancy and its association with maternal body mass index (BMI). Methods We measured 8-hydroxy-deoxyguanosine (8-OH-dG), a marker of DNA oxidative damage, in venous umbilical cord plasma from newborns of mothers with (n=34) and without (n=56) diabetes diagnoses obtained during pregnancy. Two markers of oxidative stress - namely, nitric oxide degradation products (NOx) and total glutathione (GSH) - were quantified in both mothers and newborns. The effects of BMI, glycated hemoglobin (HbA1c), age and delivery mode were also analyzed. Results Newborns of mothers with diabetes during pregnancy exhibited higher levels of 8-OH-dG than those of mothers without diabetes (P<0.001). The other markers of oxidative stress were also higher in both mothers with diabetes and their newborns, with the exception of NOx in the mothers. The association of diabetes with 8-OH-dG was independent of other analyzed factors. Conclusion The offspring of mothers with diabetes during pregnancy are born with increased genotoxicity than the offspring of mothers without diabetes. BMI and HbA1c display an independent association with 8-OH-dG, particularly in the offspring of mothers not diagnosed with diabetes.


Detection of antibody to murine cytomegalovirus by enzyme-linked immunosorbent and indirect immunofluorescence assays.
We have compared murine cytomegalovirus (MCMV) antibody determination by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay. A comparison of antibody detection with 146 serum samples at a 1:20 dilution showed 100% agreement (60 negatives and 86 positives) between the assays. There was close agreement of endpoint determinations of sera by both methods. After experimental MCMV infection, antibody to MCMV was detected by both assays as early as day 7, and high titers persisted as late as 6 months. In contrast to immunocompetent littermates, athymic nude mice did not develop antibody after infection. Mice lacking antibody detectable by ELISA were susceptible to lethal MCMV challenge. In a survey of animals from five commercial sources, MCMV antibody was not detected unless mice were experimentally infected. MCMV antibody determination by ELISA is a convenient method, comparable to the indirect immunofluorescence assay in sensitivity and specificity.


[Drug allergy among patients suffering from nasal polyps].
The study was performed in order to evaluate quantity and morphology of nasal polyps in patients with diagnosed drug allergy to aspirin, antibiotics, polocain, procain and other drugs. The study included 60 patients (27 females, 33 males aged between 18-69 years). Nasal polyp morphology was examined under electron microscope and light microscope. Morphological evaluation of epithelial and subepithelial structures was done under light microscope, taking not of inflammatory infiltration type, basal membrane, as well as degree of intestinal edema. Evaluation of epithelial cell type, their relative proportion, as well as area percentage covered by cilium, microvilli and squamous epithelium was done under electron microscope. Polyps in patients with idiosyncrasy to polish aspirin and sodium metamizole (12 patients) featured increased eosinophilic infiltration, thickened basal membrane, with elevated eosinophil percentage differential count. The epithelial percentage area was covered by squamous cells without villi. Polyps associated with idiosyncrasy to polish aspirin differed morphology compared to polyps originality from patients allergic to other medications. On the other hand polyps from patients allergic to other drugs, such as: penicillin, procainamide were morphologically (electron light microscope, light microscope) similar to those polyps caused by inflammatory factors.


[Expression of the VP2 gene of chicken infectious anemia virus in E. coli and analysis of immunogenicity].
The coding region of VP2 gene from Chicken Infectious Anemia was amplified from genome extracted from chicken liver tissue by PCR. PCR product was double digested with restriction enzymes BamH I and Sal I and cloned into pET-28a digested with BamH I and Sal I. Subsequently, the recombinant plasmid pET-28-VP2 was extracted and double digested with restriction enzymes BamH I and Sal I. After confirming its rightness by PCR and analysis of restriction endonucleases, the recombinant plasmid pET-28-VP2 was transformed into E. coli BL21 (DE3) strain. The culture was induced by 1 mmol/L IPTG at 37 degrees C for three hours and analyzed with SDS-PAGE. The result shows that gene encoding VP2 of CIAV was expressed successfully in E. coli and the fusion protein existed in supernatant, which was about 31kDa and showed specific immunoreactivity with anti-CIAV sera in Western blot. The fusion protein was purified by Ni2+ -affinity chromatography and quantitated by Bradford method. Then BALb/c mice were immunized with purified protein emulsified with Freund's complete adjuvant on day 0 and boosted twice on day 14 and 28 with the same dose of antigens emulsified with Freund's incomplete adjuvant, respectively. The serum isolated were examined by an enzyme-linked immunosorbant assay (ELISA) using the purified VP2 and CIAV as coating antigens and the serum could react with target protein and CIAV in ELISA detection test.


Racing the clock: The role of circadian rhythmicity in addiction across the lifespan.
Although potent effects of psychoactive drugs on circadian rhythms were first described over 30 years ago, research into the reciprocal relationship between the reward system and the circadian system - and the impact of this relationship on addiction - has only become a focus in the last decade. Nonetheless, great progress has been made in that short time toward understanding how drugs of abuse impact the molecular and physiological circadian clocks, as well as how disruption of normal circadian rhythm biology may contribute to addiction and ameliorate the efficacy of treatments for addiction. In particular, data have emerged demonstrating that disrupted circadian rhythms, such as those observed in shift workers and adolescents, increase susceptibility to addiction. Furthermore, circadian rhythms and addiction impact one another longitudinally - specifically from adolescence to the elderly. In this review, the current understanding of how the circadian clock interacts with substances of abuse within the context of age-dependent changes in rhythmicity, including the potential existence of a drug-sensitive clock, the correlation between chronotype and addiction vulnerability, and the importance of rhythmicity in the mesocorticolimbic dopamine system, is discussed. The primary focus is on alcohol addiction, as the preponderance of research is in this area, with references to other addictions as warranted. The implications of clock-drug interactions for the treatment of addiction will also be reviewed, and the potential of therapeutics that reset the circadian rhythm will be highlighted.


[Endometrial hyperplasia: A review].
Endometrial hyperplasias can be divided into two categories based on the presence or absence of cytological atypia and further classified as simple or complex according to the extent of architectural abnormalities. They are usually diagnosed because of irregular bleeding in perimenopause. Hysteroscopy with a biopsy gives a more accurate diagnosis than transvaginal ultrasonography, sonohysterography, or blind curettage. Endometrial hyperplasias with no cytological atypia, regarded as a response to unopposed endogenous estrogenic stimulation, are normally treated with progestins. The intra-uterine route (levonorgestrel intra-uterine system) is more effective and better tolerated than the oral route. Either conservative surgery (endometrial resection, thermal ablation) or radical surgery (hysterectomy) in the case of other genital diseases is performed on women who did not respond to medical treatment. Endometrial hyperplasias with cytological atypia, considered as intra-epithelial neoplasias, are traditionally treated by hysterectomy. The absence of management protocols in the literature offers various treatment options and indications. Gonadotropin-releasing hormone agonists, danazol, or aromatase inhibitor are effective, but have adverse effects and are expensive. Endometrial ablation can be performed as a first line therapy in women suffering from bleeding related to hyperplasia without cytological atypia. Medical treatment may be offered to young women suffered from hyperplasias with cytological atypia and desiring pregnancy.


Other Iatrogenic Immunodeficiency-Associated Lymphoproliferative Disorders with a T- or NK-cell phenotype.
Other iatrogenic immunodeficiency-associated lymphoproliferative disorders (OIIA-LPDs) with a T- or NK-cell phenotype are markedly rare, with only a limited number of cases having been reported thus far. Methotrexate (MTX) is the most common agent used for OIIA-LPD patients, and 43 cases of MTX-associated T-LPDs (MTX T-LPDs) and five cases of MTX-associated NK/T-LPDs (MTX NK-LPDs) have been described. In addition to MTX T-LPDs and MTX NK/T-LPDs, T-LPD and NK/T-LPDs have been reported in patients receiving other immunosuppressive agents such as thiopurines, TNF antagonists, and cyclosporine. Hepatosplenic T-cell lymphoma (HSTL) is specifically associated with iatrogenic immunodeficiency, and 10% of HSTL cases develop in patients receiving thiopurines and/or TNF antagonists for inflammatory bowel disease (IBD). In this review, we focused on MTX T-LPD, MTX NK/T-LPD, and HSTL in patients with IBD. These T- and NK/T-cell associated OIIA-LPDs are the most common in daily medical practice.


Estimating Secondary Organic Aerosol Production from Toluene Photochemistry in a Megacity of China.
The production of secondary organic aerosols (SOA) from toluene photochemistry in Shanghai, a megacity of China, was estimated by two approaches, the parametrization method and the tracer-based method. The temporal profiles of toluene, together with other fifty-six volatile organic compounds (VOCs), were characterized. Combing with the vapor wall loss corrected SOA yields derived from chamber experiments, the estimated toluene SOA by the parametrization method as embodied in the two-product model contributes up to ∼40% of the total SOA budget during summertime. 2,3-Dihydroxy-4-oxopentanoic acid (DHOPA), a unique product from the OH-initiated oxidation of toluene in the presence of elevated NOx, was used as a tracer to back calculate the toluene SOA concentrations. By taking account for the effect of gas-particle partitioning processes on the fraction of DHOPA in the particle phase, the estimated toluene SOA concentrations agree within ∼33% with the estimates by the parametrization method. The agreement between these two independent approaches highlight the need to update current model frameworks with recent laboratory advances for a more accurate representation of SOA formation in regions with substantial anthropogenic emissions.


NRROS Negatively Regulates Osteoclast Differentiation by Inhibiting RANKL-Mediated NF-N:B and Reactive Oxygen Species Pathways.
Negative regulator of reactive oxygen species (NRROS) is known to repress ROS generation in phagocytes. In this study, we examined the roles of NRROS in both osteoclasts and osteoblasts. Our results demonstrate that NRROS negatively regulates the differentiation of osteoclasts, but not osteoblasts. Further, overexpression of NRROS in osteoclast precursor cells attenuates RANKL-induced osteoclast differentiation. Conversely, osteoclast differentiation is enhanced upon siRNA-mediated knockdown of NRROS. Additionally, NRROS attenuates RANKL-induced NF-N:B activation, as well as degradation of the NOX1 and NOX2 proteins, which are required for ROS generation. Based on our observations, we present NRROS as a novel negative regulator of RANKL-induced osteoclastogenesis.


Hsp90 C-terminal inhibitors exhibit antimigratory activity by disrupting the Hsp90α/Aha1 complex in PC3-MM2 cells.
Human Hsp90 isoforms are molecular chaperones that are often up-regulated in malignances and represent a primary target for Hsp90 inhibitors undergoing clinical evaluation. Hsp90α is a stress-inducible isoform of Hsp90 that plays a significant role in apoptosis and metastasis. Though Hsp90α is secreted into the extracellular space under metastatic conditions, its role in cancer biology is poorly understood. We report that Hsp90α associates with the Aha1 co-chaperone and found this complex to localize in secretory vesicles and at the leading edge of migrating cells. Knockdown of Hsp90α resulted in a defect in cell migration. The functional role of Hsp90α/Aha1 was studied by treating the cells with various novobiocin-based Hsp90 C-terminal inhibitors. These inhibitors disrupted the Hsp90α/Aha1 complex, caused a cytoplasmic redistribution of Hsp90α and Aha1, and decreased cell migration. Structure-function studies determined that disruption of Hsp90α/Aha1 association and inhibition of cell migration correlated with the presence of a benzamide side chain, since an acetamide substituted analog was less effective. Our results show that disruption of Hsp90α/Aha1 interactions with novobiocin-based Hsp90 C-terminal inhibitors may limit the metastatic potential of tumors.


Discussion on pharmacogenetic interaction in G6PD deficiency and methods to identify potential hemolytic drugs.
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common form of red blood cell enzymopathy. The disorder has reached polymorphic frequencies in different parts of the world due to the relative protection conferred against malaria. G6PD is a housekeeping X-linked gene encoding the first enzyme of the pentose phosphate pathway, an NADPH-producing dehydrogenase. Because erythrocytes do not generate NADPH in any other way than pentose phosphate pathway, they are more susceptible than any other cells to oxidative damages. G6PD deficiency is a prime example of a hemolytic anemia due to an interaction between an intracorpuscular cause and an extracorpuscular cause, because in the majority of cases an exogenous agent triggers hemolysis. Hemolysis, in fact, can be caused by exposure to oxidant agents. Although studies performed on epidemiology, genetics and molecular biology have broaden the information on G6pd deficiency, there are still no reliable and validated methods to test drug hemolytic potential in G6PD deficient patients. The review gives an overview of current knowledge on G6pd deficiency and on the methods that have been developed so far in order to identify drugs causing acute hemolytic anemia in G6pd deficiency. Moreover, we discuss the new potential preclinical strategies to assess, in vitro and in vivo, drug hemolytic risks.


Plasma protein insudation as an index of early coronary atherogenesis.
Two hundred ninety-nine paraffin-embedded coronary artery blocks from 68 autopsy cases were serially sectioned. The blocks were selected to provide a range from normal through various stages of atherosclerosis, and sections were examined with the indirect immunofluorescence technique for intramural distribution of plasma albumin, fibrinogen, and immunoglobulin gamma (IgG). Cryostat-sections of 44 blocks from 22 of the same cases were examined with the same technique for distribution of apolipoprotein B. Alteration of protein insudation in the artery wall was a sensitive index of coronary atherogenesis. The sequence in which these proteins were involved in the initiation and development of early atherosclerotic lesions was analyzed by determining the average relative intimal thickness and relative lumen size that was associated with the first occurrence of altered insudation of each of these proteins. Results indicate that changed plasma albumin insudation is the earliest sign of a focal intimal lesion, and increasing albumin insudation shows the strongest association with intimal plaque growth. The other proteins tested showed altered insudation, in the order IgG, fibrinogen, apolipoprotein B. The results indicate that a progressive increase in permeability of the coronary artery endothelium occurs in the early stages of atherogenesis. Patterns of IgG localization provide evidence of both early systemic and subsequent local immune reactions being involved in atherogenesis. Altered albumin and apolipoprotein B insudation levels have stronger correlation coefficients with relative intimal thickness and relative lumen size than do those IgG and fibrinogen. The extremely high correlation coefficients shown by albumin emphasizes the importance of edema in determining plaque size and lumen stenosis.


Interleukin-29 uses a type 1 interferon-like program to promote antiviral responses in human hepatocytes.
Interleukin-28A (IL-28A), IL-28B and IL-29 are a family of class II cytokines that stimulate antiviral responses through a heterodimeric receptor that is distinct from the type I interferon (IFN) receptor. To better understand how this newly described family of cytokines regulates the antiviral state, we compared various cellular responses elicited by IL-29 and IFN-alpha. Here we show that these cytokines stimulate similar patterns of signal transducer and activator of transcription 1 (STAT-1), -2, -3, and -5 phosphorylation and nearly identical patterns of gene expression when analyzed in two distinct cell types by microarray analysis. Interestingly, the IL-29 receptor is preferentially expressed on primary hepatocytes within normal liver and pegylated forms of IL-29 and IFN-alpha induced equivalent 2'5' oligoadenylate synthetase (OAS) and MX1 gene expression in this cell type. Pegylated IL-29 also produced a significant reduction in human hepatitis B and hepatitis C viral load in vitro and reduced the cytopathic effect caused by the fully replicating flavivirus, West Nile virus. In conclusion, IL-29 and IFN-alpha stimulate identical antiviral responses despite their utilization of different receptors. This fact, combined with significant receptor expression in hepatitis virus-infected livers, suggests that IL-29 may have therapeutic value against chronic viral hepatitis in human patients.


Antifungal drug discovery and development--fourth international summit. 10-11 March 1999, Princeton, New Jersey, USA.
Genetic and genomic approaches to discovery, selection and evaluation of antifungal targets were extensively discussed by several speakers at this meeting. Experimental targets with early lead compounds available from screening programs include translation (Ribogene Inc), transcription (SCRIPTGEN Pharmaceuticals Inc), and protein geranylgeranylation (Mitotix Inc). Classes of compounds which are more advanced in preclinical evaluation include the sordarins, inhibitors of fungal elongation factor 2 under development by Glaxo Wellcome plc and Merck and Co Inc, and a series of fungicidal peptides originally derived from a domain of the human neutrophil bactericidal permeability-increasing protein (BPI, Xoma Ltd). Preclinical and early clinical data were presented for two compounds: caspofungin (MK-0991, Merck and Co Inc), an echinocandin with a broad-spectrum of activity for parenteral application, and Sch-56592 (Schering-Plough Corp), an orally active triazole. The oral antimycotic, terbinafine (Novartis AG), primarily used in dermatological infections, shows potent synergy with azoles and has potential applications against several serious and drug-resistant fungal pathogens. Amphotericin B, which has long been the gold standard for therapy of life-threatening infections, is now available in several liposomal formulations, including AmBisome (Fujisawa Pharmaceutical Co Ltd) and Abelcet (The Liposome Company Inc) which show a reduced incidence of adverse events.


Influence of age on tolerability, safety and effectiveness of subdermal contraceptive implants.
The use of long-acting contraceptive methods is on the rise. The aim of this study was to describe the main variables (effectiveness, tolerability, menstrual bleeding) associated with the use of subdermal contraceptive implants and to investigate the influence of age on these variables. This was a descriptive, retrospective, observational study of 221 cases of contraceptive implants inserted at a Spanish hospital between 2006 and 2015. The mean age of implant users was 31.2 ± 7.5 years. Effectiveness was 100% and good tolerability was recorded for 86.5%. Infrequent bleeding was the most common bleeding pattern, followed by amenorrhoea. Of the 221 implants inserted, 47.5% were removed. The main reasons were expiration (54.3%) and discomfort due to bleeding alterations and other adverse effects (25.7%). Nulliparity and weight gain were significantly associated with an increased probability of implant removal. This study shows that implants were highly effective, safe and well-tolerated in our population. The age of users had no influence on any of the study variables analysed. Impact Statement What is already known on this subject? Subdermal contraceptive implants are long-acting reversible contraceptives which are both safe and effective. What do the results of this study add? The age of users had no influence on any of the study variables analysed. Nulliparity and weight gain were significantly associated with an increased probability of implant removal. What are the implications of these findings for clinical practice and/or further research? Subdermal contraceptive implants were a safe and effective long-acting progestin contraceptive method for women from all age groups in our series because no significant age-related differences were observed for the tolerability, vaginal bleeding patterns, the effectiveness, the adverse effects or any other variables.


[Sonic hedgehog and prostate growth regulation].
Sonic hedgehog (Shh) is a key signal regulatory factor in embryonic development. It is reported that Shh signaling plays important roles in prostatic duct differentiation and matrix-epithelium interaction, and thus regulates the development, growth and cell proliferation of the prostate. A disorder in Shh signaling will lead to the production and proliferation of tumor cells. An exploration into the mechanism of Shh signaling in the normal growth and diseased condition of the prostate will offer some significant ideas for the studies on the pathogenesy of prostate diseases.


Partial molal volumes and compressibilities of phosphoric acid and sodium phosphates in 0.725 molal NaCl at 25 °C.
The apparent molal volume V(ϕ) and adiabatic compressibility κ(ϕ) of H(3)PO(4), NaH(2)PO(4), Na(2)HPO(4), and Na(3)PO(4) have been determined from density and sound speed measurements in 0.725 m NaCl solutions at 25 °C. The partial molal volumes V(i)* and compressibilities κ(i)* have been determined by extrapolating the values of V(ϕ) and κ(ϕ) to infinite dilution in the NaCl media. The results have been compared to earlier measurements made on a number of electrolytes in NaCl media. The phosphate results showed correlations between the values of V(i)* and κ(i)* as found for other electrolytes. The values for the volume ΔV(i) and compressibility Δκ(i) changes for the dissociation of H(3)PO(4) in 0.725 m NaCl have been calculated from the measurements. The calculated values of ΔV(i) and compressibility Δκ(i) have been used to estimate the effect of pressure K(P)/K(0) on the dissociation of H(3)PO(4) in the oceans using RT ln[K(P)/K(0)] = -ΔV(i)P + 0.5Δκ(i)P(2), where P is the applied pressure in bars. The results are in reasonable agreement with directly measured values.


Enchondromatosis with features of dysspondyloenchondromatosis and Maffucci syndrome.
We report a girl with multiple enchondromatosis, unequal leg length, short stature, congenital scoliosis, lymphangioma, and cutaneous hemangiomata. The skeletal findings were consistent with the clinical and radiological features of dysspondyloenchondromatosis except that short stature was not apparent in the neonatal period. Dysspondyloenchondromatosis is a rare disorder, one of the several types of multiple enchondromatosis with spinal abnormalities. In previous reports of this condition the association of vascular lesions usually found in Maffucci syndrome has not been described.


Mixed treatment comparison meta-analysis of aspirin, warfarin, and new anticoagulants for stroke prevention in patients with nonvalvular atrial fibrillation.
Warfarin and aspirin are used to prevent stroke in patients with atrial fibrillation (AF). There are inherent challenges with both treatments, including variable and inconsistent benefit and increased bleeding risks. The availability of new anticoagulants offers some alternatives. A mixed treatment comparison meta-analysis to evaluate direct and indirect treatment data including aspirin, warfarin apixaban, dabigatran, edoxaban, and rivaroxaban for the prevention of primary or secondary stroke in patients with AF. A comprehensive, systematic literature search was conducted to identify randomized trials comparing aspirin, warfarin, apixaban, dabigatran, edoxaban, and rivaroxaban in patients with AF requiring treatment for stroke prevention. Open-label and blinded designs were included if they evaluated any stroke or any bleeding event. Data on stroke and bleeding events were abstracted, verified, evaluated, scored, and entered into Aggregate Data Drug Information System version 1.16 to generate a mixed treatment comparison meta-analysis. Direct and indirect comparisons were evaluated, and we looked for inconsistency in closed loop structures. Data are reported as rate ratios with 95% credible intervals. In addition, we reviewed variance statistics and explored variance with node-splitting models. Our literature search yielded 30 articles, 21 of which were included. All treatments except aspirin reduced the risk of any stroke compared with placebo. Warfarin (0.43 [0.33-0.57]), apixaban (0.37 [0.27-0.54]), dabigatran (0.34 [0.21-0.57]), rivaroxaban (0.36 [0.22-0.60]), and aspirin with clopidogrel (0.73 [0.53-0.99]) were more protective than aspirin alone. Warfarin and the new anticoagulants were similar in the reduction of stroke, vascular death, and mortality. There was no difference in major bleeding between any treatment group. There were more nonmajor bleeding events when comparing warfarin and apixaban (1.83 [1.05-4.03]); no other differences between warfarin and the other new anticoagulants were found. This mixed treatment comparison meta-analysis found similarity between warfarin and the new anticoagulants with the exception of one comparison, in which warfarin was associated with more non-major bleeding than apixaban. Thus, the new anticoagulants are therapeutically comparable when warfarin is inappropriate.


The distally based sural artery flap for ankle and foot coverage.
The sural artery flap is a distally based fasciocutaneous flap that has many advantages to offer for coverage in the foot and ankle area. It has the largest arc of rotation of all the regional flaps and does not require sacrifice of any major artery, and moderate-to-large-sized defects can be covered adequately. The dissection technique is simple, and donor site morbidity is minimal. We report our experience with 17 cases. Age range was from 13 to 56 years. Ten (59%) defects were posttraumatic, 3 (17%) were related to reconstructive surgery of the foot or tendon Achilles', 2 (11%) resulted from tumor resection, and 1 each were from infection and gunshot wound. The smallest flap was 6 x 4 cm and the largest was 15 x 12 cm, with the average size being 11 x 7.5 cm. In 5 cases, the donor site was closed primarily, and in other cases, split-thickness skin graft was needed. The short saphenous vein was included in the pedicle in all cases. There was no incidence of complete flap necrosis. Follow-up ranged from 3 to 30 months. Two cases (12%) developed partial superficial necrosis. In 1 case, there was partial wound dehiscence that needed debridement and repair. Another case had postoperative discharge, which subsided after removal of the calcaneal plate. None of the patients complained of any functional problem related to loss of sensation along the lateral border of the foot. The sural island flap is a reliable, safe, and easy method of providing soft tissue coverage in the area of the foot and ankle.


[Blood concentrations of maprotiline in depressive patients].
Blood levels of Maprotiline were analysed and their relationship to the clinical response was examined in 89 depressed inpatients, according DSM III criteria for Major Depressive Episode, given the drug treatment for 3 weeks. Maprotiline produced marked decreases in mean MADRS and COVI scale scores by the end of treatment. On day 21, no correlation between blood levels of Maprotiline and MADRS or COVI scores were found when all patients were considered. Nevertheless, significant correlations were observed on day 14 (r = .22; p less than .05 for MADRS and r = .23; p less than .05 for COVI scale). In addition, a significant correlation between MADRS or COVI scale scores and Maprotiline blood levels were observed on days 14 and 21 in subgroups of young patients, severe depression (high scores to clinical global investigations), during of at least 3 months, treated without other drug than Maprotiline and good responders.


[Isoflavones from vines of Pueraria lobata].
To investigate the isoflavones from the vines of Pueraria lobata. The compounds were isolated by column chromatography over silica gel and RP-C18, and purified by Sephadex LH-20 column chromatography and preparative TLC. The structures were elucidated on the basis of physico-chemical properties and spectral data. Twelve compounds were isolated and identified as: 3'-methoxydaidzein (1), formononetin (2), genistein (3), daidzein (4), daidzin (5), genistin (6), ononin (7), 5-hydroxyl ononin (8), calycosin (9), 6"-O-acetyl genistein (10), 6"-O-acetyl daidzin (11), puerarin (12). For the first time, compounds 9-11 were isolated from the genus Pueraria plant, and compounds 1, 3, 6-8 were obtained from the vines of this plant.