Formaldehyde preservation for deferred measurements of alkaline phosphatase activities in marine samples.
Alkaline phosphatases are the main enzymes required by microorganisms to hydrolyse organic phosphorus into available phosphate in aquatic environments. The investigations of alkaline phosphatase activity (APA) usually generate numerous samples (size fractionation, Michaelis-Menten kinetics). Therefore, convenient and reliable preservation of incubated samples for a deferred analysis would be very useful when measurements cannot be performed right away. The APA of marine pond waters was measured using 4-Methylumbelliferyl phosphate (MUF-P) as the fluorogenic substrate modelling natural organic phosphorus compounds. Where typical inhibitors of other enzymatic activities, such as 1% sodium dodecyl sulfate, mercuric chloride, or buffered solutions of ammonium and glycine, failed to stop APA, the addition of formaldehyde efficiently inhibited APA. The effect of formaldehyde was the strongest with the highest concentration tested (4% final concentration) and in buffered (pH 8) solutions. Since a slow and gradual increase in APA may persist with time, the combination of the addition of 4% buffered formaldehyde with immediate freezing is the best method to entirely inhibit APA. The maximal rate of hydrolysis (Vmax) and the Michaelis constant (Km) of formaldehyde (4%)-inhibited samples did not significantly change during storage at -20 °C for 11 days. The method was successfully tested on samples with extremely high values of APA (15000-40000 nM h-1) that were preserved for 1 month at -20 °C (98% inhibition). This method is a reliable and useful means of preserving incubated samples, and it provides convenient controls for background fluorescence of water and substrate, without provoking abiotic hydrolysis of the substrate.


Angiogenesis: the major abnormality of the keratin-14 IL-4 transgenic mouse model of atopic dermatitis.
Angiogenesis plays an important role in psoriasis, but its role in atopic dermatitis is unknown. The authors examined the dermal microvasculature of an IL-4 transgenic mouse model of atopic dermatitis to determine whether angiogenesis was present. Transmission and scanning electron microscopy and confocal microscopy studies were performed. Transmission electron microscopy showed sprouting, transcapillary pillars of intussusception, thickened endothelial cells with large nuclei, and increased interendothelial junctional cleft number and length. Compared to nontransgenic littermates, there was a significant increase in the lengths and numbers of the interendothelial junctional clefts, along with a decrease in the length ratios of tight junction to interendothelial junctional clefts in both the early and late disease stages. In the early and late skin lesions, scanning electron microscopy of vascular corrosion casts showed disorganization of the capillary network hierarchy with increased density of capillary sprouts. Confocal microscopy of the animals with early and late skin lesions showed significant reduction in tight junction protein claudin-5. Angiogenesis is the major pathologic feature in this model of atopic dermatitis. The chronic skin inflammation is intertwined with and may cause the angiogenesis, but the angiogenesis itself is likely to be important in this disease process.


Mupirocin: a new topical antibiotic.
One hundred fifty-three strains of Staphylococcus aureus recovered from infected eczema frequently demonstrated resistance to multiple antibiotics. Penicillin and ampicillin resistance was extremely frequent (88%), methicillin resistance was found in nearly 14% of strains, and erythromycin and tetracycline resistance was present in 16%. S. aureus strains were uniformly sensitive to vancomycin, mupirocin, and cephalosporins. Experimental infections in human volunteers showed topical therapy with 2% mupirocin was more effective than oral erythromycin in suppression of both S. aureus and Streptococcus pyogenes.


Non-lytic M13 phage-based highly sensitive impedimetric cytosensor for detection of coliforms.
A highly sensitive and selective non-lytic M13 phage-based electrochemical impedance spectroscopy (EIS) cytosensor for early detection of coliforms is introduced for the first time. Gold nanoparticles were electrochemically deposited on the surface of glassy carbon electrode, and the M13 phage particles were immobilized on them using 3-mercaptopropionic acid linker and zero-length crosslinking chemistry (EDC/NHS). Next, the sensor surface was blocked to avoid non-specific binding. The M13-EIS cytosensor was tested for detection of F+ pili Escherichia coli species, using XL1-Blue and K12 strains, as examples of coliforms. The selectivity against non-host strains was demonstrated using Pseudomonas Chlororaphis. The binding of E. coli to the M13 phage on the cytosensor surface increased the charge transfer resistance, enabling detection of coliforms. The biosensor achieved a limit of detection (LOD) of 14 CFU/mL, the lowest reported to-date using EIS-phage sensors, and exhibited a high selectivity towards the tested coliforms. The SEM micrographs confirmed the successful capturing of E. coli on the M13-based EIS cytosensor. Moreover, the sensor showed almost the same sensitivity in the simulated river water samples as in phosphate buffer, reflecting its applicability to real samples. On the other hand, this sensor system exhibited high stability under harsh environmental conditions of pH (3.0-10.0) and temperature as high as 45 °C for up to two weeks. Overall, this sensor system has excellent potential for real field detection of fecal coliforms.


Computed CD4 percentage as a low-cost method for determining pediatric antiretroviral treatment eligibility.
The performance of the WHO recommendations for pediatric antiretroviral treatment (ART) in resource poor settings is insufficiently documented in routine care. We compared clinical and immunological criteria in 366 children aged 0 to 12 years in Kinshasa and evaluated a simple computation to estimate CD4 percent, based on CD4 count, total white blood cell count and percentage lymphocytes. Kappa (kappa) statistic was used to evaluate eligibility criteria and linear regression to determine trends of CD4 percent, count and total lymphocyte count (TLC). Agreement between clinical and immunological eligibility criteria was poor (kappa = 0.26). One third of children clinically eligible for ART were ineligible using immunological criteria; one third of children immunologically eligible were ineligible using clinical criteria. Among children presenting in WHO stage I or II, 54 (32%) were eligible according to immunological criteria. Agreement with CD4 percent was poor for TLC (kappa = 0.04), fair for total CD4 count (kappa = 0.39) and substantial for CD4 percent computational estimate (kappa = 0.71). Among 5 to 12 years old children, total CD4 count was higher in younger age groups (-32 cells/mm3 per year older), CD4 percent was similar across age groups. Age-specific thresholds for CD4 percent optimally determine pediatric ART eligibility. The use of CD4 percent computational estimate may increase ART access in settings with limited access to CD4 percent assays.


An isolated enriched organ-specific cancer neoantigen of human lung cancer for leukocyte adherence inhibition assays.
Until now, measurement of human antitumor immunity to organ-specific cancer neoantigen (OSN) by the leukocyte adherence inhibition (LAI) assay depended on using crude extracts of cancer. In this study, a new method is presented to generate and to isolate a highly enriched OSN from spent medium of a lung cancer cell line, NCI-H69, grown in chemically defined medium. Production of large quantities of OSN with minimal contamination by extraneous proteins was possible. Four physicochemical steps were used to give a 1000-fold enrichment of OSN activity: anion-exchange and molecular-sieve chromatography; Blue Sepharose affinity chromatography; and finally anion-exchange high-pressure liquid chromatography. The enriched OSN isolates showed dose-response antigenicity when tested in LAI assay with leukocytes from lung cancer patients but had no antigenicity with leukocytes from control subjects or patients having malignant melanoma, colon cancer, or pancreatic cancer. Cross-reactive antigenicity was observed with leukocytes from patients with breast cancer and slight reactivity with leukocytes from bladder cancer patients. The final isolate from the four-step separation procedure as well as the isolates produced using additional separation techniques consistently had antigenicity at less than 10 ng in blocking LAI and 500 ng in the direct assay and showed components with molecular weights of about 62,000 +/- 3,000 (SD) (p62), 40,000 +/- 3,000 (p40), and 25,000 +/- 1,000 (p25) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The OSN isolates on two-dimensional gels showed p40 to have microheterogeneity (seven spots), with a pl from 6.2 to 7.6, and p62 and p25 as even more basic streaks. The polypeptide bearing the antigenic determinant was not purified, although we tried to separate p62, p40, and p25 to determine whether they carried the OSN determinant. The results of this study are important in showing that an isolate of an organ-specific tumor antigen containing 5 to 13 components, as determined by highly sensitive silver stains and radiolabeled patterns on single and two-dimensional gels, can be used successfully in LAI to measure tumor immune responses.


Genome sequence of a highly prevalent porcine partetravirus in Mainland China.
Partetravirus is a novel defined genus of animal parvoviruses. Here, we first report the genome sequence of porcine partetravirus strain JSNJ62, which is highly prevalent in mainland China. It will help in understanding the epidemiology and molecular characteristics of the porcine partetravirus.


Comparing methods for estimating leaf area index by multi-angular remote sensing in winter wheat.
The reflectance of wheat's canopy exhibits angular sensitivity, which can influence the accuracy of different methods for its leaf area index (LAI) estimation through multi-angular remote sensing. The primary objective of this study was to assess and compare the ability of various methods for LAI estimation from 13 view zenith angles (VZAs). The four methods included: (1) common hyper-spectral vegetation indices (VIs), (2) optimal two-band combination VIs (i.e., VIs: normalized difference index, simple ratio index, and difference vegetation index), (3) back-propagation neural network (BPNN), and (4) partial least squares regression (PLSR). Our results demonstrated that the red-edge plays a key role in estimating LAI, in that the traditional VIs, optimal two-band VIs, and PLSR including the red-edge band all showed satisfactory performance, with coefficient of determination (R2) > 0.72 in the nadir direction. However, the estimation accuracy of LAI was not positively related with band number, and BPNN gave unsatisfactory results under a larger viewing angle, with R2 ≤ 0.60 for extreme angles. The predictive ability of all four methods declined with an increasing VZA, with reliable LAI estimation near the nadir direction. Importantly, by comparing the four methods, PLSR emerged as superior in both its estimation accuracy and angular insensitivity, with R2 = 0.83 in the nadir direction and ≥ 0.65 for extreme angles. For this reason, we highly recommend it be used with multi-angular remote sensing data, especially in agricultural applications.


Trade-offs between grain yields and ecological efficiencies in a wheat-maize cropping system using optimized tillage and fertilization management on the North China Plain.
Optimized fertilizer and tillage management can be an effective strategy for high ecological efficiencies as well as crop yields. The objective of this study was to assess the impact of diverse management practices on carbon footprint, and ecosystem services in a wheat-maize cropping system. An in situ field experiment field was conducted from 2018 to 2020 on the North China Plain, and six treatments were established: deep tillage (DT), shallow tillage (ST), no tillage (NT), deep tillage + adding organic fertilizer (DTF), shallow tillage + adding organic fertilizer (STF), and no tillage + adding organic fertilizer (NTF). The results showed that adding organic fertilizer and the deeper tillage depth caused higher direct CO2 and N2O emission fluxes. DTF treatment significantly increased carbon footprint either per-unit area (CFa) or per-unit net income (CFe). Compared with DT treatment, STF treatment had higher CFa but lower CFe by increasing net income through boosted crop yields. Besides, the highest ecosystem service values (ESV) were present in STF treatment during both 2 years (42,017.13 CNY ha-1 and 43,352.03 CNY ha-1). In conclusion, STF treatment was an optimal management practice to trade-off grain yields and ecological efficiencies in a wheat-maize cropping system. Furthermore, this study highlights that adding organic fertilizer could be an efficient option toward sustainable farmland utilization with high soil carbon sequestration capacity and high ESV.


Inhaled medication usage in post-menopausal women and lifetime tobacco smoke exposure: The Women's Health Initiative Observational Study.
While active smoking is a causal agent in respiratory disease, the independent role of secondhand smoke (SHS) merits further investigation. We investigated associations between lifetime active smoking and exposure to secondhand smoke - studied independently - and current use of 1 or more inhaled medications as a surrogate for prevalent pulmonary disease in post-menopausal women. Information on lifetime active and passive tobacco exposure and inhaled pulmonary medication usage at enrollment was collected from 88,185 postmenopausal women aged 50-79 enrolled in the Women's Health Initiative Observational Study from 1993 to 1998 at 40 centers in the United States. Participants were recruited from localities surrounding the study centers using a variety of methods, including informational mailings and mass media campaigns. Multivariate adjusted regression models were used to estimate odds ratios and 95% CI according to levels of active smoking and SHS exposure, and trends were tested across categories. Ever active smokers had an overall OR of 1.97 (95% CI 1.58-2.45) for having one or more prescribed inhaled medication compared with never-smoking women not exposed to active or passive smoke. The overall OR for using inhalers for never-smoking women exposed to any SHS compared with the same reference group was 1.33 (95% CI 1.07-1.65). In a quantified analysis of SHS, never-smoking women with the highest levels of lifetime SHS exposure had an estimated risk of inhaled medication usage of 1.74 (95% CI 1.32-2.30). The risk of requiring one or more prescribed inhaled medications for pulmonary disease was significantly higher in post-menopausal women who ever smoked or who had lifetime exposure to SHS.


[Benign disease of the throat in xerotomograms (Part I) (author's transl)].
The advantages of xeroradiography in radiology of the soft parts are emphasized. The technique in tomography of the throat is described. Typical benign diseases of the throat like acute or chronic inflammation, edema, pareses of the vocal cords and injuries are demonstrated on xerotomograms and discussed. Detail of the structures of the throat is better seen in xeroradiographic tomograms than in Röntgentomograms.


Localization of NADPH-diaphorase activity in the dental pulp, periodontium and alveolar bone of the rat.
In this study we examined the presence and localization of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity in the dental pulp, periodontal tissues and alveolar bone of the rat. The presence of NADPH-d activity was also examined in cat pulp. The rat histochemical analysis revealed the presence of prominent NADPH-d activity both in cells of the sub-odontoblastic cell layer and in the odontoblasts, in the root as well as in the coronal pulp regions. In the pulpal horns, odontoblasts often had long processes with a high level of labelling indicating NADPH-d activity extending through the predentin and dentin. Moreover, endothelial cells of pulpal blood vessels were positive for NADPH-d in both species. However, no clearcut examples were found of pulpal nerve fibres positive for NADPH-d in the rat or cat and denervation performed in rats did not alter the enzyme staining patterns. In the periodontal tissue, NADPH-d activity was localized to cells on the alveolar bone surface of the periodontal ligament and, in addition, alveolar bone marrow crypts were filled with intensely labelled cells. In the gingival papillae, NADPH-d activity was observed in the basal cell layer of the epithelium. Endothelial cells of periodontal and gingival blood vessels showing positive staining for NADPH-d were occasionally noted.


Mixotrophic lifestyle of the ichthyotoxic haptophyte, Prymnesium parvum, offered different sources of phosphorus.
Many harmful algae are mixoplanktonic, i.e. they combine phototrophy and phagotrophy, and this ability may explain their ecological success, especially when environmental conditions are not optimal for autotrophic growth. In this study, laboratory experiments were conducted with the mixotrophic and ichthyotoxic haptophyte Prymnesium parvum (strain CCAP 946/6) to test the effects of phosphorus (P) sufficiency and deficiency on its growth rate, phagotrophic and lytic activities. P-deficient P. parvum cultures were grown without or with addition of P in the form of inorganic phosphorus (nutrients) and/or living algal prey (i.e. the cryptophyte Teleaulax amphioxeia). The ingestion rate of P. parvum and prey mortality rate were calculated using flow cytometry measurements based on pigment-derived-fluorescence to distinguish between prey, predators and digesting predators. The first aim of the study was to develop a method taking into account the rate of digestion, allowing the calculation of ingestion rates over a two-week period. Growth rates of P. parvum were higher in the treatments with live prey, irrespective of the concentration of inorganic P, and maximum growth rates were found when both inorganic and organic P in form of prey were added (0.79 ± 0.07 d-1). In addition, the mortality rate of T. amphioxeia induced by lytic compounds was 0.2 ± 0.02 d-1 in the P-deficient treatment, while no mortality was observed under P-sufficiency in the present experiments. This study also revealed the mortality due to cell lysis exceeded that of prey ingestion. Therefore, additional experiments were conducted with lysed prey cells. When grown with debris from prey cells, the mean growth rate of P. parvum was similar to monocultures without additions of prey debris (0.30 ± 0.1 vs. 0.38 ± 0.03 d-1), suggesting that P. parvum is able to grow on prey debris, but not as fast as with live prey. These results provide the first quantitative evidence over two weeks of experiment that ingestion of organic P in the form of living prey and/or debris of prey plays an important role in P. parvum growth and may explain its ecological success in a nutrient-limited environments.


Hate speech and abusive language detection in Indonesian social media: Progress and challenges.
Nowadays Hate Speech and Abusive Language (HSAL) have spread extensively over social media. The easy use of social media allows people to abuse the media to spread HSAL. Hate speech and abusive language in social media must be detected because they can trigger conflict among citizens. Not only in social media, but HSAL also often trigger conflict in real life. In recent years, many scholars have researched HSAL detection in various languages and media. However, there are still many tasks on HSAL detection that need to be done to develop a better HSAL detection system. This paper discusses a summary of Indonesian HSAL detection research, conducted by utilizing the Kitchenham systematic literature review method. Based on our summary, we found that most Indonesian HSAL research still uses the classic machine-learning approach with classic text representation features that experimented on the Twitter text dataset. We also found several challenges and tasks that need to be addressed to build a better HSAL detection system in Indonesian social media that can detect the hate speech target, category, and levels; and the hate speech buzzer, thread starter, and fake account spreader.


Assay and purity control of oxytetracycline and doxycycline by thin-layer chromatography--a comparison with liquid chromatography.
A thin-layer chromatographic (TLC) method using densitometry is described for the assay and purity control of oxytetracycline and doxycycline. With a mobile phase of dichloromethane-methanol-water (59:35:6, v/v/v) and a silica gel thin-layer, previously sprayed with 10% sodium edetate solution adjusted to pH 9.0, all the potential impurities of oxytetracycline or doxycycline are well separated from the main components and from each other. Results obtained with TLC are compared with those obtained by previously established liquid chromatography (LC) methods using poly(styrene-divinylbenzene) stationary phases. A good correlation was obtained (r greater than 0.9999). For TLC the relative standard deviation (RSD) for the assay of the main component was less than 2%, for LC the RSD was less than 1%.


Gender and Racial Diversity among U.S. Urology Residency Programs and Efforts to Improve It: You Can't Be What You Can't See.
The U.S. urology workforce lacks ethnic and gender diversity. Few programs exist to increase diversity, and little is known on their effectiveness. We assessed the landscape of specific programs designed to increase underrepresented in medicine (URiM) and female student participation in the U.S. Urology Match, and understand the concerns and attitudes of those students. To better understand urology-specific programs, we sent an 11-item survey to all 143 urology residency programs. To better understand the concerns and attitudes of URiM and female students participating in the U.S. Urology Match, we sent a 12-item survey to the students who participated in the Match from 2017 to 2021. Lastly, we evaluated trends in match rate using Match data from 2019 to 2021. Among programs, 43% responded to our survey. Most residency programs offer a wide array of initiatives to increase their diversity, with unconscious bias training being the most frequent (78.7%). Programs with at least 1 female faculty member were associated with increased recruitment of female residents over time (p=0.047). A similar trend was seen in programs with URiM faculty. Among students, 10.5% responded to our survey, of whom 79.2% were unaware of any programs at their institution geared toward URiM or female students. Match data revealed that women were more likely to match (p=0.002), and URiM students were less likely to match (p <0.001) compared to the overall Match rate. Urology programs are making substantial efforts to improve diversity, but the message is lacking reach. Having a diverse faculty did make a difference in programs' ability to diversify.


Spiked nanorosette: A novel zinc oxide-gold textured nanostructure for impedance biosensing of target DNA signatures.
The creation of nanostructure is profound for the generation of nanobiosensors in several medical diagnosis. Here, we employed an aqueous hydrothermal route using Zinc-oxide (ZnO) and Gold (Au), which under optimal conditions formed an ultra-crystalline rose-like nanostructure textured with nanowires on the surface, coined as "spiked nanorosette." The spiked nanorosette structures was further characterized to possess crystallites of ZnO and Au grains with average sizes of 27.60 and 32.33 nm, respectively. The intensity for both ZnO (002) and Au (111) planes of the nanocomposite was inferred to be controlled by fine-tuning the percentage of Au nanoparticles doped in the ZnO/Au matrix, as referred by X-ray diffraction analysis. The formation of ZnO/Au-hybrid nanorosettes were additionally verified by the distinct corresponding peaks from photoluminescence and X-ray photoelectron spectroscopy, supported by electrical validations. The biorecognition properties of the spiked nanorosettes were also examined using custom targeted and non-target DNA sequences. The DNA targeting capabilities of the nanostructures were analyzed by Fourier Transform Infrared and electrochemical impedance spectroscopy. The fabricated nanowire-embedded nanorosette exhibited a detection limit at the lower picomolar range of 1 × 10-12 M, with high selectivity, stability and reproducibility and good linearity, under optimal conditions. Impedance-based techniques are more sensitive to the detection of nucleic acid molecule whereas this novel spiked nanorosette demonstrate promising attributes as excellent nanostructures for nanobiosensor developments and their potential future application for nucleic-acids or disease diagnostics.


The long-term results of electric fire hand burns in children.
An 11-year retrospective series of 95 deep, electric fire hand burns in children was studied and the epidemiology, treatment and operative management analysed. From this series a group of 14 patients was recruited to assess the long-term results of this injury. At long-term follow-up (mean 14.4 years) the recorded parameters included range of movement, hand sensation, power and size. Photographs were taken and any subjective problems noted. Analysis of these data showed a marked variation in range of movement, sensation, size and appearance of the injured hands. Despite this, complaints about cosmetic results or any restriction of activities were rare. In conclusion, these injuries are particularly destructive but with appropriate surgical management, albeit protracted, and despite imperfect cosmetic results, excellent function can be obtained in the growing hand.


Interstitial brachytherapy technique for chest wall refractory recurrence of breast cancer.
To report the treatment effect of interstitial brachytherapy for chest wall locoregional recurrence of breast cancer. This 44-year-old female presented with chest wall recurrence seven years after modified radical mastectomy for stage II breast cancer. Despite external beam radiation and chemotherapy, the lesion expanded as 5.3 × 5.1 × 3.0 cm(3), and 8.0 × 5.1 × 4.0 cm(3). The locoregional recurrent tumor was treated with interstitial brachytherapy under ultrasound guidance. The brachytherapy dose was 30 Gy in 6 fractions of 5 Gy each. Removal of the recurrent tumor was securely achieved by interstitial brachytherapy guided with ultrasound scanning. The refractory tumor in patient healed uneventfully after interstitial brachytherapy without recurrence during the 7 months of follow-up. The ultrasound-guided interstitial brachytherapy may be effective for refractory recurrence of breast cancer.


Extrusion 3D printing of nutraceutical oral dosage forms formulated with monoglycerides oleogels and phytosterols mixtures.
Among the potential applications of 3D printing, the development of products with personalized characteristics in the area of food and nutraceuticals represents an important field that must still be explored. The aim of this work was to evaluate the production of nutraceutical oral forms by extrusion-based 3D printing (E3DP) using mixtures of monoglycerides (MG) oleogels and phytosterols (PS) as printing materials. These materials were obtained using MG (10 or 20%wt), high oleic sunflower oil, and variable amounts of PS (20-50%wt PS/oleogel). An ad-hoc extrusion 3D printer composed of a heated syringe and a cooling build platform was used. Rheological tests were carried out to determine the mixtures gel point, in order to select appropriate printing temperatures, as well as the yield stress of the final materials. Hardness of printed forms was obtained by compression tests. Additionally, oral forms were produced by manual extrusion using molds for comparison. It was found that oral forms were successfully printed when using mixtures containing a maximum of 30 and 40%wt PS/oleogel for oleogels formulated with 10 and 20%wt of MG, respectively. Moreover, the best printed forms corresponded to the mixtures with the lowest gelation temperatures. These printed forms were structurally stable, with uniform weight and shape, and maximum hardness of 12.55 N. Hardness values of printed oral forms did not show a correlation with those obtained by manual extrusion using molds, indicating that this parameter was affected by solid composition, cooling rate, and the fragility generated for layers superposition. In conclusion, it was demonstrated that mixtures of MG oleogels and PS can be used for E3DP production of nutraceutical oral forms suggesting that oleogels have excellent potential as materials able to incorporate liposoluble active ingredients to be used as extrusion printing materials.


[Structure elucidation of butyl 6alpha-fluoro-11beta-hydroxy-16alpha-methyl-3,20-dioxo-1,4-pregnadien-21-oate (fluocortin butylester) (author's transl)].
The structure of butyl-6alpha-fluoro-11beta-hydroxy-16alpha-methyl-3,20-dioxo-1,4-pregnadien-21-oate (fluocortin butylester, Vaspit) was elucidated by the spectrometric methods UV, IR, NMR and MS.


Modeling of a single nanoparticle interaction with the human blood plasma proteins.
When nanoparticles are introduced into a physiological environment, proteins and lipids immediately cover their surface, forming a protein "corona". It is well recognized that the corona structure influences the biological response of the body. Two deterministic models for corona formation of the human blood serum proteins around a single nanoparticle are presented and studied numerically in this paper. One of them is based on a coupled system of PDEs and involves diffusion of proteins toward the nanoparticle surface. The other one is described by ODEs and is a limit version of the first model as the protein diffusivity tends to infinity. The protein diffusivity influence on the temporal corona structure is studied in detail. Results are presented using figures and discussion.


Nitric oxide reacts with the single-electron reduced active site of cytochrome c oxidase.
The reduction kinetics of the mutants K354M and D124N of the Paracoccus denitrificans cytochrome oxidase (heme aa(3)) by ruthenium hexamine was investigated by stopped-flow spectrophotometry in the absence/presence of NO. Quick heme a reduction precedes the biphasic heme a(3) reduction, which is extremely slow in the K354M mutant (k(1) = 0.09 +/- 0.01 s(-1); k(2) = 0.005 +/- 0.001 s(-1)) but much faster in the D124N aa(3) (k(1) = 21 +/- 6 s(-1); k(2) = 2.2 +/- 0.5 s(-1)). NO causes a very large increase (>100-fold) in the rate constant of heme a(3) reduction in the K354M mutant but only a approximately 5-fold increase in the D124N mutant. The K354M enzyme reacts rapidly with O(2) when fully reduced but is essentially inactive in turnover; thus, it was proposed that impaired reduction of the active site is the cause of activity loss. Since at saturating [NO], heme a(3) reduction is approximately 100-fold faster than the extremely low turnover rate, we conclude that, contrary to O(2), NO can react not only with the two-electron but also with the single-electron reduced active site. This mechanism would account for the efficient inhibition of cytochrome oxidase activity by NO in the wild-type enzyme, both from P. denitrificans and from beef heart. Results also suggest that the H(+)-conducting K pathway, but not the D pathway, controls the kinetics of the single-electron reduction of the active site.


Flexibility and distraction after monosegmental and bisegmental lumbosacral fixation with angular stable fixators.
In human lumbosacral spines, the flexibility and intersegmental distraction allowed by four monosegmental and bisegmental intrapedicular devices during compression and flexion/compression loading were investigated. To compare the flexibility and intersegmental distraction allowed by four monosegmental and bisegmental intrapedicular, lumbosacral fixation devices applied to destabilized cadaveric spines, and to determine the effect of each device on the flexibility and intersegmental distraction of the motion segment above each fused segment. The lumbosacral segment is the most mobile region in the lumbar spine, exhibiting the highest range of motion in both flexion and extension. Therefore, the fixation of this and the adjacent segment is of special clinical interest. L5-S1 facetectomy or L5 laminectomy procedures were performed on cadaveric human lumbosacral spines. Fixation devices then were applied across one or two levels, and intersegmental motion under flexion/compression or under pure compression loads was monitored. The flexibility and posterior strain allowed by the four implants did not differ significantly between implants. The strain across the stabilized site (monosegmental and bisegmental) was below 10% for all devices tested. The flexibility and distraction were reduced to levels below intact after the fixators were applied. The distraction across the segment above the fusion was not increased because of fixation with these four fixators. Angular stable fixation devices, such as those described here, provide adequate stabilization of the posteriorly destabilized spine.


Caregiving self-efficacy and pain assessment by family caregivers of people living with dementia.
Like other older adults, people living with dementia (PLWD) experience pain, and the task of pain assessment often falls to family caregivers. In this study, we surveyed family caregivers of PLWD to determine the frequency with which they use different elements of pain assessment. We also determined correlations of family caregivers' characteristics (caregiving self-efficacy, relationship duration and type, mood, education level, and health literacy) with their use of the elements of pain assessment. Participants reported frequent use of many pain assessment elements. Statistically significant correlations were found between caregiving self-efficacy for obtaining respite and asking others about noticed behavior change (rho=.0.41, p=.007); and for responding to disruptive patient behaviors for multiple pain assessment elements including observing pain behaviors (rho=0.49, p<.001), asking others about noticed behavior change (rho=0.54, p<.001) and rechecking (rho=0.56, p<.001). Continued efforts are needed to describe pain assessments by family caregivers of PLWD.


Obsessive compulsive disorder and psychopathic behaviour in Babylon.
The history of obsessive compulsive, phobic and psychopathic behaviour can be traced to the 17th century AD. We draw attention to these behaviours in a Babylonian cuneiform medical text known as Shurpu. These three categories were united in the Babylonian mind around the concept of the māmīt 'oath' idea, the behaviour habits being so unbreakable it appeared that the subject had sworn an oath to do or not to do the action involved. The behavioural accounts were entirely objective, including what we would call immature, antisocial and criminal behaviour, and obsessional categories of contamination, aggression, orderliness of objects, sex and religion. They do not include subjective descriptions of obsessional thoughts, ruminations or the subject's attitude to their own behaviour, which are more modern fields of enquiry. The Babylonians had no understanding of brain or psychological function but they were remarkable describers of medical disease and behaviour. Although they had both physical and supernatural theories of many medical disorders and behaviours, they had an open mind on these particular behaviours which they regarded as a 'mystery' yet to be 'resolved'. We are not aware of comparable accounts of these behaviours in ancient Egyptian or classical medicine. These Babylonian descriptions extend the history of these disorders to the first half of the second millennium BC.


Silence and related symptoms in children and adolescents: a network approach to selective mutism.
Silence in certain situations represents the core symptom of selective mutism (SM). However, it is unclear what additional symptoms are part of this disorder. Although knowledge of symptoms is essential for diagnostics and intervention, to date, only scarce research exists on circumscribed symptoms of SM. Given the large overlap between SM and social anxiety disorder (SAD), it remains also unclear which symptoms can differentiate both disorders. A network analysis of potential symptoms of SM was performed based on a mixed sample of N = 899 children and adolescents with and without indication of SM (n = 629 with silence in certain situations). In a preliminary analysis, we demonstrated that children with and without silence in certain situations do not differ with respect to their network structure, justifying an analysis on the entire mixed sample. Possible communities (symptom clusters) within the network and thus potential latent variables were examined, and symptoms were analyzed in terms of their centrality (the extent to which they are associated with other symptoms in the network). To investigate the differentiability of symptoms of the SM network from symptoms of SAD, we computed a network that additionally contains symptoms of SAD. In the resulting network on symptoms of SM, silence was, as expected, the symptom with the highest centrality. We identified two communities (symptom cluster): (1) symptoms associated with the fear response of freezing, (2) symptoms associated with speech production and avoidance. SM network symptoms and SAD symptoms largely formed two separate symptom clusters, with only selectivity of speaking behavior (more talkative at home and taciturn or mute outside the home) falling into a common cluster with SAD symptoms. Silence appears to have been confirmed by analysis as a core symptom of SM. Additional anxiety-related symptoms, such as avoidance behavior or motor inhibition associated with freezing, seem to co-occur with silence. The two communities of SM potentially indicate different mechanisms of silence. The symptoms of SM appear to be distinguishable from those of SAD, although there seems to be overlap in terms of difficulty speaking in situations outside the home.


Effects of cerium and tungsten substitution on antiviral and antibacterial properties of lanthanum molybdate.
Powders of cerium (Ce)-substituted and tungsten (W)-substituted La2Mo2O9 (LMO) were prepared using polymerizable complex method. Their antiviral and antibacterial performances were then evaluated using bacteriophage Qβ, bacteriophage Φ6, Escherichia coli, and Staphylococcus aureus. The obtained powders, which were almost single-phase, exhibited both antiviral and antibacterial properties. Effects of dissolved ions on their antiviral activity against bacteriophage Qβ were remarkable. A certain contribution of direct contact to the powder surface was also inferred along with the dissolved ion effect for antiviral activity against bacteriophage Φ6. Dissolved ion effects and pH values suggest that both Mo and W are in the form of polyacids. Antiviral activity against bacteriophage Φ6 was improved by substituting Ce for La in LMO. Similarly to LMO, Ce-substituted LMO exhibited hydrophobicity. Inactivation of alkaline phosphatase enzyme proteins was inferred as one mechanism of the antiviral and antibacterial activities of the obtained powders.


A simple and rapid immunochromatographic test strip for detection of pathogenic isolates of Vibrio harveyi.
Mouse monoclonal antibodies (MAbs) and rabbit polyclonal antibody (PAb) against Vibrio harveyi were generated from immunization of mice and rabbits with highly virulent isolate of V. harveyi. Two MAbs specific to virulent isolates of V. harveyi were obtained and one of them (VH4) was selected to conjugate with colloidal gold as the detector antibody was laid on a sample pad. Rabbit polyclonal antibody was used as the capture antibody at the test line (T) and goat anti-mouse IgG antibody (GAM) was used as the capture antibody at the control line (C) of nitrocellulose strip. The ready-to-use strip was held in a plastic case and then stored in a desiccated plastic bag. A sample volume of 100 microl of bacterial suspension from various sources mixed with application buffer was applied to the sample chamber at one end of the strip and allowed to flow by chromatography through the nitrocellulose membrane to the other end. In test samples containing virulent isolates of V. harveyi, the bacteria would bind to the monoclonal antibody conjugated with colloidal gold and the resulting complex would be captured by the antibodies at the test line to give a reddish-purple band. Any unbound monoclonal antibody conjugated with colloidal gold moved across the test line would be captured by the GAM and form a band at the control line (C). In sample without V. harveyi or with V. harveyi below the limit (<10(6) CFU/ml) of detection for the kit, only the control line band was observed. If the test sample was pre-enriched in tryptic soy broth (TSB) for 6 h before application to the strip, the sensitivity would increase to 1-10 CFU/ml which is comparable to that of PCR. This method could be used to detect pathogenic isolates of V. harveyi in pond water or infected shrimp in order to monitor and to reduce the risk of V. harveyi outbreak in the shrimp culture. The beneficial features of this kit are that simple, convenient and quick results (within 15 min) can be obtained without the requirement of sophisticated tools or special equipments and skills.


Characterization of the genotoxic action of three structurally related 1,2-dihaloalkanes in Drosophila melanogaster.
The genetic activity profiles of three structurally related dihaloalkanes, 1,2-dibromoethane (DBE), 1,2-dichloroethane (DCE) and 1-bromo-2-chloroethane (BCE), were compared in germ cells and somatic tissue of Drosophila melanogaster. The two genotoxicity indices estimated after mutagen exposure of male germ cells were (i) the hypermutability index fexr-/fexr+, measured by the increased frequency of induced recessive lethals (RL) in a strain defective in DNA excision repair (exr-), as compared to the wild type (exr+); (ii) the relative clastogenicity index CL/RL, expressed by the ratio of chromosomal aberrations (CL; ring-X loss) to RL determined in exr+ strains. The fexr-/fexr+ index for DBE was 4-5 times higher than those for DCE and BCE, suggesting a difference in the types of premutagenic lesions produced by DBE in comparison to DCE and BCE. The relative clastogenicity indices for BCE (CL/RL = 0.29) and DCE (0.41) are similar to the value of 0.37 estimated for DBE in an earlier study, all indicating that the three compounds or their metabolites are incapable of forming DNA crosslinks. In somatic cells, after inhalation treatment of female larvae, the effectiveness for the induction of interchromosomal recombination decreased in the order BCE > or = DBE > DCE. It is concluded that in accordance with other studies also in Drosophila the glutathione-mediated pathway is the major cause of genotoxicity caused by DBE, DCE and BCE.


Improving horizontal plane locomotion via leg angle control.
The lateral leg spring model has been shown to accurately represent horizontal plane locomotion characteristics of sprawled posture insects such as the cockroach Blaberus discoidalis. While passively stable periodic gaits result from employing a constant leg touch-down angle for this model, utilizing a similar protocol for a point mass model of locomotion in three dimensions produces only unstable periodic gaits. In this work, we return to the horizontal plane model and develop a simple control law that prescribes variations in the leg touch-down angle in response to external perturbations. The resulting control law applies control once per stance phase, at the instant of leg touch-down, and depends upon previous leg angles defined in the body reference frame. As a result, our control action is consistent with the neural activity evidenced by B. discoidalis during locomotion over flat and rough terrain, and utilizes variables easily sensed by insect mechanoreceptors. Application of control in the lateral leg spring model is shown to improve stability of periodic gaits, enable stabilization of previously unstable periodic gaits, and maintain or improve the basin of stability of periodic gaits. The magnitude of leg touch-down angle variations utilized during stabilization appear consistent with the natural variations evidenced by single legs during locomotion over flat terrain.


Ultrarapid freezing reveals that skeletal muscle caveolae are semipermanent structures.
Rat myofibers in the resting state and under imposed conditions of moderate to severe physiological, pharmacological, and mechanical stress were prepared by ultrarapid freezing and examined by freeze fracture. In rapidly frozen rat myofibers, caveolae morphology and distribution were found to be unchanged by brief or prolonged rest, brief direct electrical stimulation and concomitant contractile activity, prolonged direct electrical stimulation (to fatigue), myofiber stretch (within normal myofiber limits), or careful compressive scission. However, caveolae were greatly reduced or eliminated in number and size by severe mechanical disruption (shredding and/or tearing) of myofibers. Thus, we conclude that unlike apparently similar surface specialization in other cell types, skeletal muscle caveolae are not transient stages in a caveolae----vesicle endocytotic-exocytotic cycle, nor are they a membrane reservoir for normal stretch/contractile activity. Rather, they are (semi)permanent structures in the muscle plasma membrane with as yet undetermined function and kinetics.


Fecal microbiota transfer for bowel disorders: efficacy or hype?
Dysbiosis has been related to the pathophysiology of disorders of - gut-brain interaction (DGBI) including irritable bowel syndrome (IBS) and functional constipation (FC). Accordingly, modulation of gut microbiota has been proposed as a potential treatment for these disorders. Gut microbiota modulation can be effected by probiotics, prebiotics, symbiotics, postbiotics, antibiotics and fecal transplantation (FMT) or bacteriotherapy. The latter is currently used for recurrent or severe Clostridium difficile colitis and has been the focus of recent research in IBS and FC. Several case series reported promising results for FMT in patients with IBS and FC, which prompted the conduction of randomized controlled trials (RCT) in these DGBI. Both case series and RCTs are herein discussed. To the best of our knowledge, as of yet, 5 RCTs have been published on IBS and one in FC with slow colonic transit. In IBS, the majority of studies have used the IBS severity scoring system (IBS-SSS) as an outcome measure; however, the selection criteria were different among the trials as well as the route and form of administration of the FMT. Therefore, the results are inconsistent and no conclusion can be drawn. Some studies suggest that the presence of post-infection (PI)-IBS and the baseline microbiota status in the donors could be predictor factors of successful FMT in IBS. In constipation with slow colonic transit, the FMT seems to be more effective, although the data is based on only one RCT. We believe that larger RCTs, controlled with true placebos and considering baseline intestinal microbiota of the study subjects as well as donors' microbiota are still needed before recommending FMT in IBS and/or FC. History of previous GI infection (e.g. PI-IBS) and IBS subtypes should also be taken into account.


Mapping of second messenger and rolipram receptors in mammalian brain.
Autoradiographic localizations of major second messengers and a selective cyclic adenosine monophosphate (cyclic-AMP) phosphodiesterase in the brain were visualized in the gerbil and the rat using receptor autoradiography. [3H]Phorbol 12,13-dibutyrate (PDBu), [3H]inositol 1,4,5-trisphosphate (IP3), [3H]forskolin, [3H]cyclic-AMP, and [3H]rolipram were used to label protein kinase C, IP3 receptor, adenylate cyclase, cyclic-AMP-dependent protein kinase (cyclic-AMP-DPK), and Ca2+/calmodulin-independent cyclic-AMP phosphodiesterase (PDE), respectively. Most second messengers and rolipram binding activities were especially found in the limbic system, basal ganglia, and cerebellum. Marked differences were noted in the hippocampus, where cyclic-AMP and rolipram binding activities were very low in gerbils but high in rats. In contrast, regional localization in the binding sites of PDBu, IP3, and forskolin in gerbil brain was relatively similar to that in rat brain. Further, alteration of the cyclic-AMP and rolipram binding sites was studied in the gerbil hippocampus 7 days after 10-min cerebral ischemia. The results suggest that the gerbil differs from the rat with respect to the characteristic neurons or interneurons, especially in the hippocampal formation. This finding may help further elucidate the relationship or difference between gerbils and rats for brain function and behavioral pharmacology. Furthermore, our results suggest that cyclic-AMP and rolipram binding sites are predominantly distributed on the pyramidal cell layer of the hippocampal CA1 sector and that transient cerebral ischemia can cause marked reduction in these binding sites in the hippocampus.


Sodium lactate reversal of electrophysiological effects of imipramine in guinea-pig ventricular myocardium.
Overdose cardiac effects of imipramine are due to fast Na channel blockade and are clinically reversed by administration of sodium lactate which induces alkalosis (about pH 7.50) and hypernatremia (about 8 mM). The mechanisms of this beneficial effect of Na lactate were explored in vitro on guinea-pig ventricular myocardium using the microelectrode technique. The time-course effects of the clinically relevant concentration of 10 microM imipramine on action potential characteristics were examined at pH 7.20 and pH 7.50. To test whether alkalinisation per se is important or whether an increase in Na concentration plays a major role in the reversal effect, preparations were exposed to increasing concentrations (1, 3, 10, 30, 100 mM) of either Na lactate, bicarbonate or chloride in the absence or in the presence of 10 microM imipramine at pH 7.50. The influence of elevating osmolality was evaluated with equivalent concentrations of sucrose. Imipramine alone significantly depressed Vmax and shortened action potential duration at all phases of repolarisation. All three high sodium solutions reversed imipramine effects. However the reversal effect was already obvious with 10 mM Na lactate and 10 mM NaHCO3 but not 10 mM NaCl. Osmolality did not reverse the imipramine-induced Vmax depression. The results suggest that at the clinically relevant 10 mM concentration, sodium lactate and bicarbonate may displace imipramine from its receptor site on the Na channel by causing alkalosis at the membrane level without profoundly affecting the driving force of the Na current, whereas at the upper concentrations, the increase in Na ion concentrations is predominantly involved in the reversal of imipramine effects.


Expression of leucocyte antigens by cells from the metrial gland of the pregnant rat.
The function of the metrial gland of the rat, and particularly of its characteristic population of granulated cells, remains unknown. However, several lines of evidence suggest that the granulated cells may derive from lymphocytes, and play a role in the immunology of pregnancy. In this study, antigen expression by granulated and other cell populations from the metrial glands of rats at Days 13 and 14 of pregnancy was studied by an indirect immunoperoxidase method. Acetone-fixed frozen sections, and cytocentrifuge preparations of collagenase-dispersed metrial gland tissue in which numbers of granulated cells had been increased by density-gradient centrifugation, were used. The primary antibodies used recognised, inter alia, B lymphocytes (MRC OX-3, MRC OX-6, MRC OX-12), T lymphocytes (MRC OX-8, W3/25, MRC OX-19), neutrophils (MRC OX-42) and cells of the monocyte/macrophage series (MRC OX-3, MRC OX-6, MRC OX-42, MRC OX-43). The majority of the granulated cells, including smaller, "immature" forms, were unlabelled by any of these antibodies. Some lymphocytes, and varying numbers of larger, non-granulated cells, were labelled by OX-6, OX-12, W3/25, OX-42 and OX-43. In addition to lymphocytes, labelled cells included neutrophils (OX-42), endothelial cells (OX-43), and probably some macrophages (OX-6, OX-43). OX-12, which recognises the kappa chain of rat IgG, labelled some large cells which may have been stromal cells. These findings do not support the concept that the granulated cells are derived from lymphocytes.


New indole-7-aldehyde derivatives as melatonin analogues; synthesis and screening their antioxidant and anticancer potential.
Over the last decade, there has been substantial interest in the use of melatonin (MLT) and MLT-like compounds in the treatment of several diseases. MLT can scavenge different reactive oxygen species and can also stimulate the synthesis of antioxidant enzymes. Our ongoing study relies on changing the groups in the different modifiable sites of the indole ring to increase the antioxidant activity. In this study a new approach for substitution of indole ring as indole based MLT analogue was proposed. We report the synthesis and characterization of a series of new indole-7-aldehyde hydrazide/hydrazone derivatives as indole-based MLT analogues. Anticancer potential of the compounds were evaluated both by their antioxidant and CYP1 inhibitory activities. In vitro antioxidant capacity of the compounds was investigated both in a cell-based (DCFH assay) and a cell-free (DPPH assay) assay. Potential inhibitory effects of the compounds on CYP1 catalytic activity were investigated via EROD assay. Cytotoxic activity of the compounds was further evaluated by the MTT assay in CHO-K1 cells. MLT analogues having an o-halogenated aromatic moiety exhibited effective antioxidant properties without having any cytotoxic effect. In conclusion, MLT derivatives represent promising scaffolds for discovery of effective antioxidant agents.


Theoretical investigation on the low-lying electronic states of diatomic magnesium bismuthide including the spin-orbit coupling effect.
High-level ab initio computations have been performed to investigate molecular structures, potential energy curves, vibrational energy levels and spectroscopic constants for twelve Λ-S states of the first four dissociation limits of MgBi. Characterizations of seven Ω states, corresponding to the first and the second Λ-S dissociation limits, have been explored for the first time. The spin-orbit coupling effect is revealed to have introduced a significant impact on the pattern of these electronic states and interactions among them. Our predictions for molecular structures and spectroscopic constants of MgBi are compared with available data of other magnesium-group 18 family species. Regular tendencies of these parameters are clearly exhibited when the group 18 atom is replaced by another one in the group. Information associated with transition dipole moments, Franck-Condon factors, vibrational branching ratios and radiative lifetimes between the Ω states are obtained and their transitional properties are analyzed and discussed. The results and data determined in this work are expected to guide and assist laboratorial detections of MgBi and to extend our understanding for the magnesium-group 18 species.


Gastric hamartomatous polyp without polyposis coli: radiologic diagnosis.
Gastric hamartomatous polyps were found in 25 patients over a period of 4 1/2 years, for an incidence of 11% in all endoscopically biopsied polyps. The number of polyps was fewer than 5 in most patients. All polyps were sessile, sharply demarcated, hemispheric protrusions, measuring up to 10 mm in diameter but most were less than 5 mm. The most characteristic finding differing from hyperplastic polyps, adenoma, and polypoid carcinoma was that hamartomatous polyps were located in the fundic gland mucosa, which was demonstrated as a rugal area on moderately distended double-contrast radiographs.


The primary structure of the lymphocytic choriomeningitis virus L gene encodes a putative RNA polymerase.
The complete RNA sequence of the L protein gene of lymphocytic choriomeningitis virus (LCMV) is presented. It is the first L protein sequence to be obtained for the Arenaviridae, a family of single-stranded RNA viruses which includes Lassa fever virus, and the Tacaribe complex viruses such as Pichinde and the Argentine and Bolivian hemorrhagic fever viruses. It is the largest open reading frame on the L RNA spanning 6633 nucleotides and coding for a 2210 amino acid protein with a calculated molecular weight of 254,529. Antipeptide sera identify a gene product encoded on the L RNA: it has a mass of approximately 200,000 Da and is found in virions and ribonucleoprotein complexes from infected cells (M. Singh, F. Fuller-Pace, M. J. Buchmeier, and P. J. Southern, 1987, Virology, 161, 448-456). Mutations mapped to the L gene affect plaque morphology (Kirk et al., 1980), the lethality of a virulent LCMV strain on guinea pigs (Y. Riviere, R. Ahmed, P. J. Southern, M. J. Buchmeier, and M. B. A. Oldstone, 1985, J. Virol., 55, 704-709), and the ability of a variant strain of LCMV to suppress the cytotoxic T-cell response and initiate persistent infection (M. Salvato, E. Shimomaye, P. Southern, and M. B. A. Oldstone, 1988, Virology, 164, 517-522; Ahmed et al., 1988). All of these phenotypes indicate that the viral genes on the L strand are critical elements controlling virus replication and the pattern of LCMV infection. The L gene sequence encodes a viral polymerase although this protein bears little resemblance to the published sequences of other RNA virus polymerases. Therefore the LCMV polymerase likely represents a distinct category of viral transcriptase.


Determinants of partial and adequate maternal health services utilization in Nigeria: analysis of cross-sectional survey.
Access to health services during pregnancy, childbirth and the period after birth provides a substantial opportunity to limit cases of maternal mortality. In sub-Saharan Africa, the proportions of women who utilize health services remain below 70%. This study examined the factors associated with partial and adequate maternal health services utilization in Nigeria. This paper used data from 2018 Nigeria Demographic and Health Survey (DHS) comprising 21,792 women aged 15-49 years who had given births within five years of the survey. The study focused on antenatal care attendance, place of birth and postnatal care using a combined model. Multinomial logistic regression was applied in the analysis. About 74% of the women attended antenatal care, 41% gave birth in health facilities and 21% attended postnatal care. While 68% of the women partially utilized health services, 11% adequately utilized the services. The odds of partially and adequately utilizing health services increased for ever married women, women with secondary or higher education, from richest households, living in urban area, having no problem either getting permission to visit health facility or reaching health facility. This study has revealed the factors associated with partial and adequate utilization of maternal health services in Nigeria. Such factors include education, household wealth, marital status, employment status, residence, region, media exposure, getting permission to use health service, unwillingness to visit health facility without being accompanied and distance to health facility. Efforts aimed at improving maternal health services utilization should place emphasis on these factors.


Recovery of hippocampal cholinergic activity by transplantation of septal neurons in AF64A treated rats.
Embryonic septal neurons were transplanted into the hippocampus of adult rats which had received lateral-ventricular administration of AF64A, a cholinergic neurotoxin, and the effects on hippocampal cholinergic activity were studied. One week after AF64A administration, we injected dissociated septal cell suspension into the dorsal hippocampus, unilaterally. About 3 months after the transplantation, acetylcholine (ACh)-rich septal grafts formed extensive acetylcholinesterase (AChE)-positive fibers into the host hippocampus, recovering choline acetyltransferase (ChAT) level only in the grafted side. These results indicate that septal implants can produce a partial recovery of the cholinergic activity in the chemically damaged hippocampus.


Characterization of a 95 kilodalton membrane glycoprotein associated with multi-drug resistance.
Over-expression of a 95-kDa membrane protein (P-95) has been reported previously in the multi-drug-resistant (MDR) breast cancer cell line MCF-7/AdrVp and the MDR small cell lung cancer line NCI-H1688. We have now developed anti-sera against gel-purified }-95 protein from each of these cell lines. Western blotting with each serum demonstrates a broad band at 95-kDa with detergent-solubilized membrane proteins from MCF-7/AdrVp and NCI-H1688 cells, which is barely detectable in membrane proteins from drug-sensitive parental MCF-7 cells. Each anti-serum cross-reacts with a 190-kDa membrane protein (P-190) in NCI-H1688 but not MCF-7/AdrVp cells. Immunoblotting and silver staining of NCI-H1688 membrane proteins separated by two-dimensional gel electrophoresis demonstrates that P-95 and P-190 run as broad streaks with low iso-electric points. Incubation of NCI-H1688 cells with tunicamycin or cleavage of carbohydrate residues of NCI-H1688 or MCF-7/AdrVp membrane proteins with PNGase F leads to the appearance of a sharp 35-kDa band reactive with anti-P-95 antisera. This 35-kDa protein has been isolated by two-dimensional gel electrophoresis. Neuraminidase digestion converts P-95 to a broad 65-kDa immunoreactive band, indicating the presence of terminal sialic acid residues. In conclusion, P-95 is an N-linked sialoglycoprotein with a 35-kDa polypeptide core.


The Effect of Bariatric Surgery on Urinary Incontinence in Women.
Little is known regarding the effect of bariatric surgery on urinary incontinence. Between September 2008 and November 2014, 240 female patients underwent bariatric surgery. The prevalence of urinary incontinence preoperatively was 45 % (108). Eighty-two (76 %) completed urinary function questionnaires pre-operatively and post-operatively. Fifty-seven (70 %) underwent laparoscopic gastric bypass, twenty-four (29 %) underwent sleeve gastrectomy and one underwent a banding procedure. Thirty-one (38 %) reported leaking on sneezing or coughing-stress urinary incontinence (SUI). Thirteen (16 %) complained of leaking before reaching the toilet-overactive bladder syndrome (OAB). The remaining thirty-eight (46 %) reported mixed symptoms. The mean pre-operative weight and BMI were 133 (18) kg and 50 (SD = 6.2) kg/m(2) respectively. The mean post-operative BMI drop was 16 (SD = 5.2) kg/m(2). Preoperatively, 61 (75 %) reported moderate to very severe urinary incontinence compared to 30 (37 %) post-operatively (χ (2) = 3.24.67, p = 0.050). Twenty-seven (33 %) patients reported complete resolution of their urinary incontinence. Fifty-one (62 %) patients required incontinence pads on a daily basis pre-operatively, compared to 35 (43 %) post-operatively (χ (2) = 22.211.6, p = 0.00). The mean International Consultation on Incontinence Questionnaire- Urinary Incontinence short form (ICIQ-UI SF) score was 9.3 (SD = 4.4) pre-operatively compared to 4.9 (SD = 5.3) post-operatively (t = 7.2, p = 0.000). The improvement score post-operatively was 8 (SD = 3). A significant difference in the ICIQ-UI SF was identified between OAB and SUI groups when adjusting for age, number of children, type of delivery and pre-op BMI (t = 1.98, p = 0.05). Bariatric surgery results in a clinically significant improvement in urinary incontinence. However, this is not proportional to pre-operative BMI, weight loss, age, parity and mode of delivery.


Introduction of a thermophile-sourced ion pair network in the fourth beta/alpha unit of a psychophile-derived triosephosphate isomerase from Methanococcoides burtonii significantly increases its kinetic thermal stability.
Hyperthermophile proteins commonly have higher numbers of surface ionic interactions than homologous proteins from other domains of life. PfuTIM, a triosephosphate isomerase (TIM) from the hyperthermophile archaeon, Pyrococcus furiosus, contains an intricate network of 4 ion pairs in its 4th beta/alpha unit, (β/α)4, whereas MbuTIM, a triosephosphate isomerase from a psychrophile archaeon, Methanococcoides burtonii, lacks this network. Notably, (β/α)4 is the first element of the structure formed during folding of certain TIM-type (beta/alpha)8 barrel proteins. Previously, we have shown that elimination of PfuTIM's ion pair network in PfuTIM significantly decreases its kinetic structural stability. Here, we describe the reciprocal experiment in which this ion pair network is introduced into MbuTIM, to produce MutMbuTIM. Recombinant MbuTIM displays multi-state unfolding with apparent Tm values of autonomous structural elements approaching, or above, 70°C, when a temperature scanning rate of 90°C/h is used. The protein displays significant intrinsic kinetic stability, i.e., there is a marked temperature scan rate-dependence of the Tm values associated with unfolding transitions. The Tm values drop by as much as ~10°C when the temperature scanning rate is lowered to 5°C/h. MutMbuTIM, incorporating PfuTIM's ion pair network, shows significantly higher apparent Tm values (raised by 4-6°C over those displayed by MbuTIM). MutMbuTIM also displays significantly higher kinetic thermal stability. Thus, it appears that the thermal stability of triosephosphate isomerase can be increased, or decreased, by either enhancing, or reducing, the strength of ion pair interactions stabilizing (β/α)4, presumably through reduced cooperativity (and increased autonomy) in unfolding transitions.


Wafer-scale nanocracks enable single-molecule detection and on-site analysis.
Large-area surface-enhanced Raman spectroscopy (SERS) sensing platforms displaying ultrahigh sensitivity and signal uniformity have potentially enormous sensing applicability, but they are still challenging to prepare in a scalable manner. In this study, silver nanopaste (AgNPA) was employed to prepare a wafer-scale, ultrasensitive SERS substrate. The self-generated, high-density Ag nanocracks (NCKs) with small gaps could be created on Si wafers via a spin-coating process, and provided extremely abundant hotspots for SERS analyses with ultrahigh sensitivity-down to the level of single molecules (enhancement factor: ca. 1010; detection limit: ca. 10-18 M)-and great signal reproducibility (variation: ca. 3.6%). Moreover, the Ag NCK arrays demonstrated broad applicability and practicability for on-site detection when combined with a portable 785 Raman spectrometer. This method allowed the highly sensitive detection of a diverse range of analytes (benzo[a]pyrene, di-2-ethylhexyl phthalate, aflatoxins B1, zearalenone, ractopamine, salbutamol, sildenafil, thiram, dimethoate, and methamidophos). In particular, pesticides are used extensively in agricultural production. Unfortunately, they can affect the environment and human health as a result of acute toxicity. Therefore, the simultaneous label-free detection of three different pesticides was demonstrated. Finally, the SERS substrates are fabricated through a simple, efficient, and scalable process that offers new opportunities for mass production.


The antifungal activity of graphene oxide-silver nanocomposites.
Graphene oxide (GO)-based nanocomposites' antibacteria activity exhibits great potential in clinical application. Herein we reported for the first time the preparation and enhanced antifungal activity of carbon nanoscrolls (CNSs) filled with silver nanoparticles (AgNPs). The nanoscrolls filled with silver nanoparticles were prepared by sonication, TEM picture showed AgNPs filled and wrapped inside prepared nanocomposites, the antifungal test showed that CNSs-AgNPs exhibited ideal lengthened activities against Candida albicans and Candida tropical compared with the GO-AgNPs nanocomposites based on silver nanoparticles directly deposited on the surface of grapheme oxides, which is caused by CNS-AgNPs' controlled durative slow-releasing of silver ion. It is also observed that graphene oxides exhibited no antifungal activity. In conclusion, the carbon nanoscrolls composed of graphene oxides and silver nanoparticles own enhanced and lengthened antifungal activity, and have great potential in applications such as clinical nosocomial infections and local antifungal therapy.


Comparison of the fatty acid compositions in intraepithelial and infiltrating lesions of the cervix: part I, total fatty acid profiles.
In the first part of this study, the possible role of essential total fatty acids and their metabolites during cervical carcinogenesis was investigated. Since membrane lipids play a key role in cell proliferation and differentiation, disturbances in the fatty acid compositions of cell membranes and the modulation of membrane fatty acid compositions received attention in several in vitro studies. There are, however, no reported studies where the actual total and free (unesterified) fatty acid compositions have been determined during the different stages of cervical carcinogenesis. In part I of this ex vivo study, the total fatty acid compositions of normal tissue, intraepithelial and infiltrating lesions of the cervix were compared. The fatty acid profiles that were determined make it possible to speculate about the metabolic pathways followed during cervical carcinogenesis. Lipids were extracted from biopsies of normal tissue (n=36), cervical intraepithelial lesions (n=47) and infiltrating lesions (n=47). Samples, from which the total fatty acid compositions were determined, were saponified, methylated and analysed by gas liquid chromatography (GLC). Essential fatty acid deficiency (EFAD) in the intraepithelial lesions, compared with normal tissue (linoleic acid, P< 0.01), and infiltrating lesions, compared with intraepithelial lesions (linoleic acid and arachidonic acid, P< 0.01) were observed. High levels of oleic acid were also observed when infiltrating lesions were compared with normal tissue (P< 0.01). This EFAD in cancer cells may result in many defective cell mechanisms. Although there are many risk factors for cervical cancer, the human papilloma virus has emerged over the past decade as the leading candidate to be an aetiological factor. There is ample evidence that human viral infections are associated with reduced levels of linoleic acid and thus participate in the depletion of essential fatty acids in cancer cells.


Reconstruction of large phylogenetic trees: a parallel approach.
Reconstruction of phylogenetic trees for very large datasets is a known example of a computationally hard problem. In this paper, we present a parallel computing model for the widely used Multiple Instruction Multiple Data (MIMD) architecture. Following the idea of divide-and-conquer, our model adapts the recursive-DCM3 decomposition method [Roshan, U., Moret, B.M.E., Williams, T.L., Warnow, T, 2004a. Performance of suptertree methods on various dataset decompositions. In: Binida-Emonds, O.R.P. (Eds.), Phylogenetic Supertrees: Combining Information to Reveal the Tree of Life, vol. 3 of Computational Biology, Kluwer Academics, pp. 301-328; Roshan, U., Moret, B.M.E., Williams, T.L., Warnow, T., 2004b. Rec-I-DCM3: A Fast Algorithmic Technique for reconstructing large phylogenetic trees, Proceedings of the IEEE Computational Systems Bioinformatics Conference (ICSB)] to divide datasets into smaller subproblems. It distributes computation load over multiple processors so that each processor constructs subtrees on each subproblem within a batch in parallel. It finally collects the resulting trees and merges them into a supertree. The proposed model is flexible as far as methods for dividing and merging datasets are concerned. We show that our method greatly reduces the computational time of the sequential version of the program. As a case study, our parallel approach only takes 22.1h on four processors to outperform the best score to date (Found at 123.7h by the Rec-I-DCM3 program [Roshan, U., Moret, B.M.E., Williams, T.L., Warnow, T, 2004a. Performance of suptertree methods on various dataset decompositions. In: Binida-Emonds, O.R.P. (Eds.), Phylogenetic Supertrees: Combining Information to Reveal the Tree of Life, vol. 3 of Computational Biology, Kluwer Academics, pp. 301-328; Roshan, U., Moret, B.M.E., Williams, T.L., Warnow, T., 2004b. Rec-I-DCM3: A Fast Algorithmic Technique for reconstructing large phylogenetic trees, Proceedings of the IEEE Computational Systems Bioinformatics Conference (ICSB)] on one dataset. Developed with the standard message-passing library, MPI, the program can be recompiled and run on any MIMD systems.


Physical mapping of cleavage sites recognized by restriction endonucleases on the genome of bacteriophage T5.
The DNA of bacteriophage T5 has been treated with restriction endonucleases EcoRI, HindIII, BamI, SmaI, PstI, SalI, KpnI and the electrophoretic pattern obtained in agarose gel has been analyzed in order to localize the specific cleavage sites on the T5 DNA. The localization of cleavage sites has been reduced from the electrophoretic pattern of double and partial digests, the digests of isolated restriction fragments and the digests of deletion mutant T5st(o) DNA. Four BamI cleavage sites have been found and localized on the physical map of T5 DNA at 0.21, 0.225, 0.685 and 0.725 fractional length. Endonuclease SmaI cleaves at 0.39, 0.59 and 0.69 fractional length. Endonuclease PstI cuts T5 DNA at 11 sites: 0.090, 0.210, 0.320, 0.510, 0.635, 0.670, 0.705, 0.770, 0.815, 0.840, 0.875 fractional length. Six KpnI cleavage sites have been mapped at 0.170, 0.215, 0.525, 0.755, 0.830, 0.850 fractional length. A complete cleavage map of the phage genome is presented for seven restriction enzymes.


A new perspective to assess relaxation time of tumor.
The dissimilarities in spin-lattice relaxation time (T1) of water proton from one tumor to the other is due to the yield of hypoxic fraction in the tumor. The yield of higher relaxation time (T1) in the KHT tumor compared to the RIF-1 tumor as a function of growth is due to the hypoxic fraction of the tumor. The hypoxic fraction in the tumor is also responsible for the P-31 spectral parameter changes.


Occult recurrence of monomorphic epitheliotropic intestinal T-cell lymphoma and the role of MATK gene expression in diagnosis.
Monomorphic epitheliotropic intestinal T-cell lymphomas (MEITL), formerly Type II enteropathy-associated T-cell lymphomas (EATL), are rare peripheral T-cell lymphomas. They are associated with poor survival outcomes, in part because of their late diagnosis. Although MEITLs may be reliably diagnosed based on histological and immunophenotypic findings, overlaps with other NK/T and T-cell lymphomas may confound the diagnosis. The distinctive high-level nuclear staining of the novel marker Megakaryocyte-associated tyrosine kinase (MATK) in MEITLs is an invaluable tool in distinguishing MEITL from classical EATL and other NK/T or T-cell lymphomas. 18-Fluorine-2-fluorodeoxyglucose positron emission tomography (18 F-FDG PET) has been shown to be a useful tool in the staging and follow-up of aggressive lymphomas. Herein, we describe an unusual case of occult hepatic recurrence of MEITL that was non-avid on 18 F-FDG PET, in which diagnosis was confirmed based on the expression of MATK in tumour cells. Copyright © 2016 John Wiley & Sons, Ltd.


Retrovirus-mediated gene therapy for human hepatocellular carcinoma transplanted in athymic mice.
Gene therapy using a retrovirus vector carrying herpes simplex virus thymidine kinase gene under the control of the 0.3-kb human alpha-fetoprotein (AFP) gene promoter (LNAF0.3TK virus) in combination with ganciclovir (GCV) treatment was performed in athymic mice harboring AFP-producing HuH-7 human hepatoma cells. GCV treatment resulted in pronounced growth inhibition of the virus-infected HuH-7 xenograft in mice, but did not affect growth of the parental xenograft. These results indicate that the AFP gene promoter sequence allows enough therapeutic gene expression to induce the GCV-mediated cytotoxicity in vivo in AFP-producing human hepatoma cells.


Bladder and Bowel Dysfunction in Korean Children with Down Syndrome and Parental Quality of Life.
Children with Down syndrome (DS) are more likely to experience bladder bowel dysfunction (BBD) than typically developing children, which could in turn have a serious effect on children with DS and on their parents and other family members. This study aimed to explore the prevalence of BBD in Korean children with DS and its effect on parental quality of life (QOL). To assess BBD and parental QOL, we used self-administered questionnaires (Dysfunctional Voiding Symptom Score [DVSS], Rome IV criteria, and World Health Organization Quality of Life scale [WHOQOL-BREF]) for parents of children with DS. We collected data from 86 parents between September and October 2017 through an online community website. DVSS was elevated in 26.7% of the children with DS. Specifically, 14% had daytime urinary incontinence, and 33.7% had functional constipation. Moreover, 18.6% of children had BBD according to the DVSS and Rome IV criteria. The sensitivity and specificity of DVSS to functional constipation was 55.17% and 87.72%, respectively. The BBD score and total parental QOL score were statistically correlated (r = 0.291, p = 0.007). Although children with DS are a high-risk group for BBD, their BBD symptoms are often overlooked because of their intellectual disability. Consequently, this could negatively affect children's and family's health and QOL in the long term. Health-care providers should reconsider a routine check-up of BBD in children with DS. If a child has BBD, health-care providers should consult a urologist to determine the appropriate diagnosis and intervention.


Bioremediation of petroleum hydrocarbon contaminated soil by microorganisms immobilized on sludge modified by non-ionic surfactant.
The immobilization of microorganisms on high-quality and inexpensive carriers to remediate oil-contaminated soil is an effective strategy for contaminated soil remediation. Due to the abundance in nutrients, large specific surface area, and fewer pathogens, the composting sludge is considered a high-quality immobilized material. Herein, two non-ionic surfactants, TW-80 and sophorolipid, were used to modify composted sludge. High-efficiency petroleum hydrocarbon-degrading bacteria groups selected in the laboratory were fixed on the modified composting sludge under optimal conditions. The immobilized material was placed in the soil contaminated by petroleum hydrocarbons at an additive amount of 2wt/%, and a simulated remediation experiment was performed for 90 days. Both soil properties and microbial structure were characterized. Surfactant-modified compost sludge enhances the adsorption capacity to petroleum hydrocarbon. The immobilized microorganisms in the modified compost sludge showed a good effect on the remediation of soil contaminated by petroleum hydrocarbons. In addition, immobilized materials also increase the diversity of the microbial community structure in the soil. High-efficiency petroleum hydrocarbon-degrading bacteria immobilized on surfactant-modified compost can effectively promote the degradation of petroleum hydrocarbons in the soil and increase the abundance of microorganisms in the soil. It shows the feasibility of eco-friendly remediation of hydrocarbon-contaminated soil.


Which physical factors determine tricuspid regurgitation jet area in the clinical setting?
The visual assessment of jet area has become the most common method used in daily clinic practice to evaluate valvular regurgitation. Despite the high prevalence of tricuspid regurgitation, however, few studies have systematically compared TR jet areas with a quantitative standard. To evaluate this, 40 patients in sinus rhythm with tricuspid regurgitation were analyzed: 16 with centrally directed free jets and 24 with impinging wall jets. The size of the maximal planimetered color jet area (cm2) was compared with parameters derived using the pulsed Doppler 2-dimensional echocardiographic method: regurgitant fraction and the flow convergence method (peak flow rate, effective regurgitant orifice area and momentum). Mean tricuspid regurgitant fraction averaged 33 +/- 15%, peak flow rate 76 +/- 54 cm3/s, effective regurgitant orifice area 27 +/- 21 mm2 and momentum 21,717 +/- 15,014 cm4/s2. An average of 4-chamber, and long- and short-axis areas in free jets correlated well with regurgitant fraction (r = 0.81, p < 0.001), better with peak flow rate (r = 0.94, p < 0.001), effective regurgitant orifice (r = 0.92, p < 0.001) and momentum (r = 0.94, p < 0.001). The correlation was worse, but still significant, in wall jets. For the same peak flow rate, wall jets were 75% of the size of a corresponding free jet. Jet area measurement is a good semiquantitative tool to measure tricuspid regurgitation in free jets, which correlates well with regurgitant fraction and better with new parameters available from analysis of the proximal acceleration field. In patients with eccentrically directed wall jets the correlation with planimetered jet area was worse, but still significant.


Association between tissue stress reaction and ACE2/TMPRSS2 expression in endometria of reproductive aged women before and during Covid-19 pandemic.
A potential concern has been raised regarding fertility and reproductive outcome during the Covid-19 pandemic with growing stress and anxiety. However, information on the association between tissue stress reaction and expression profiles of SARS-CoV-2 viral entry proteins, ACE2 and TMPRSS2, in endometria collected from women before (pre-pandemic) and during the Covid-19 pandemic (in-pandemic) is unknown. We aim to investigate the relationship between the expression of stress-reactive proteins and of ACE2 and TMPRSS2 in endometria collected from women during these two different time frames. We retrospectively retrieved tissue blocks of endometrial samples from 25 women in 2019 (pre-pandemic) and 25 women in 2020 (in-pandemic) who underwent hysterectomy for different gynecological indications. Immunohistochemical analysis was performed with endometrial tissue samples that were collected before and during the pandemic, using respective antibodies targeting ACE2/TMPRSS2, ADRB2 and NK1R (stress and anxiety receptor markers, respectively). The quantification of immunoreactive cells for each marker was calculated by the immunoreactive score (IRS) analysis. This retrospective cohort study was limited to small sample size. No significant differences in the IRS of ACE2 and TMPRSS2 were found between the endometria that were collected before and during the pandemic with a lack of correlation between ACE2 and TMPRSS2 expression in respective endometria (r = 0.11, pre-pandemic; r = 0.04, in-pandemic). The immunostaining levels of stress marker, ADRB2 were significantly higher in the endometria of in-pandemic group (p = 0.015) comparing to that of pre-pandemic group. Pearson's correlation coefficient analysis showed a significant correlation in the expression between ADRB2 and TMPRSS2 (r = 0.41, p = 0.042) in the endometria of in-pandemic group but not in the pre-pandemic group. The rise in stress and anxiety among women during current pandemic may elicit substantial amount of tissue stress reaction with consequent increase in the expression of SARS-CoV-2 viral entry proteins in their endometria. A lack of correlation between ACE2 and TMPRSS2 expression in endometria may reassure women in their reproductive age that they are not more susceptible to infection by SARS-CoV-2 and suggest that stressful women during this pandemic can safely decide to conceive naturally or by artificial reproductive technology.


Phenotypic complementation of the SV40 tsA mutant defect in viral DNA synthesis following microinjection of SV40 T antigen.
African green monkey cells (CV-1P) were microinjected with highly purified SV40 T antigen using protein-loaded red cell ghosts and polyethylene glycol as fusagen. The microinjected cells were infected with a temperature-sensitive mutant of SV40 (tsA209) which is defective in the initiation of viral DNA synthesis. Using in situ hybridization as an assay method, we found that PEG-microinjection of both partially and highly purified T antigen resulted in an increase in the amount of viral DNA sequences in the monolayer. Moreover, 3H-thymidine-labeled and unlabeled Hirt supernatant from microinjected, tsA209-injected cells contained significantly more SV40 DNA than comparable extracts from sham-injected, tsA209-infected or uninfected cells, which were tested in parallel. Thus the introduction of highly purified, "large" SV40 T antigen led to phenotypic complementation of the tsA defect in viral DNA synthesis.


Effects of AT1 receptor antagonism on kainate-induced seizures and concomitant changes in hippocampal extracellular noradrenaline, serotonin, and dopamine levels in Wistar-Kyoto and spontaneously hypertensive rats.
In the management of epilepsy, AT1 receptor antagonists have been suggested as an additional treatment strategy. A hyperactive brain angiotensin (Ang) II system and upregulated AT1 receptors are implicated in the cerebrovascular alterations in a genetic form of hypertension. Uncontrolled hypertension could also, in turn, be a risk factor for a seizure threshold decrease and development of epileptogenesis. The present study aimed to assess the effects of the selective AT1 receptor antagonist ZD7155 on kainic acid (KA)-induced status epilepticus (SE) development and accompanying changes in the hippocampal extracellular (EC) neurotransmitter levels of noradrenaline (NAD), serotonin (5-HT), and dopamine (DA) in spontaneously hypertensive rats (SHRs) and their parent strain Wistar-Kyoto (WKY) rats, since monoamines are well-known neurotransmitters involved in mechanisms of both epilepsy and hypertension. Status epilepticus was evoked in freely moving rats by a repetitive intraperitoneal (i.p.) administration of KA in subconvulsant doses. In the treatment group, ZD7155 (5mg/kg i.p.) was coadministered with the first KA injection. Spontaneously hypertensive rats exhibited higher susceptibility to SE than WKY rats, but the AT1 receptor antagonist did not alter the development of SE in SHRs or in WKY rats. In vivo microdialysis demonstrated significant KA-induced increases of the hippocampal NAD and DA levels in SHRs and of NAD, 5-HT, and DA in WKY rats. Although SHRs developed more severe seizures while receiving a lower dose of KA compared to WKY rats, AT1 receptor antagonism completely prevented all KA-induced increases of hippocampal monoamine levels in both rat strains without affecting seizure development per se. These results suggest a lack of direct relationship between KA-induced seizure susceptibility and adaptive changes of hippocampal NAD, 5-HT, and DA levels in the effects of ZD7155 in WKY rats and SHRs.


Timing of parturition in two species of viviparous lizard: influences of beta-adrenergic stimulation and temperature upon uterine responses to arginine vasotocin (AVT).
The southern snow skink Niveoscincus microlepidotus is a viviparous alpine lizard with biennial reproduction, in which embryos are fully developed before winter but parturition is delayed until spring. We aimed to determine whether, in this species, in vitro uterine preparations are responsive to arginine vasotocin (AVT) and prostaglandin (PGF(2alpha)) in autumn and spring, and whether pre-treatment with the beta-adrenergic agonist isoproterenol decreases the effectiveness of AVT in stimulating uterine contractions. Using the spotted snow skink (Niveoscincus ocellatus), an annually breeding species, we aimed to determine influences of temperature and the beta-adrenergic system upon the response to AVT in vivo. In both N. microlepidotus and N. ocellatus females are more responsive to AVT than to PGF(2alpha), and that the response to AVT is decreased, but not prevented, by beta-adrenergic stimulation. In N. microlepidotus, uteri are equally responsive in both seasons to the hormones administered. In N. ocellatus environmental conditions, specifically, temperature, modulate the response to AVT in vivo with the time to parturition increasing as temperature decreases. We conclude that in these viviparous squamates the endocrine cascade leading to parturition is modulated by the beta-adrenergic system, and that this may reflect the mechanism by which the timing of parturition is tied to suitable environmental conditions.


Functional inhibition of c-Myc using novel inhibitors identified through "hot spot" targeting.
Protein-protein interactions drive various biological processes in healthy as well as disease states. The transcription factor c-Myc plays a crucial role in maintaining cellular homeostasis, and its deregulated expression is linked to various human cancers; therefore, it can be considered a viable target for cancer therapeutics. However, the structural heterogeneity of c-Myc due to its disordered nature poses a major challenge to drug discovery. In the present study, we used an in silico alanine scanning mutagenesis approach to identify "hot spot" residues within the c-Myc/Myc-associated factor X interface, which is highly disordered and has not yet been systematically analyzed for potential small molecule binding sites. We then used the information gained from this analysis to screen potential inhibitors using a conformation ensemble approach. The fluorescence-based biophysical experiments showed that the identified hit molecules displayed noncovalent interactions with these hot spot residues, and further cell-based experiments showed substantial in vitro potency against diverse c-Myc-expressing cancer/stem cells by deregulating c-Myc activity. These biophysical and computational studies demonstrated stable binding of the hit compounds with the disordered c-Myc protein. Collectively, our data indicated effective drug targeting of the disordered c-Myc protein via the determination of hot spot residues in the c-Myc/Myc-associated factor X heterodimer.


Analysis of purines and pyrimidines by mixed partition-adsorption normal-phase high-performance liquid chromatography.
This paper summarizes the results in the development of mixed partition-adsorption (MPA) normal-phase high-performance liquid chromatography published in the last 10 years. The MPA normal-phase systems are an alternative approach not only to the adsorption normal-phase mode but also to the most widely used reversed-phase mode in the separation area of purine and pyrimidine derivatives. It is shown that the MPA systems are applicable in analytical practice.


N6-methyladenosine (m6A) RNA methylation mediated by methyltransferase complex subunit WTAP regulates amelogenesis.
N6-methyladenosine (m6A) RNA methylation, one of the most widespread posttranscriptional modifications in eukaryotes, plays crucial roles in various developmental processes. The m6A modification process is catalyzed by a methyltransferase complex that includes Wilms tumor 1-associated protein (WTAP) as a key component. Whether the development of dental enamel is regulated by m6A RNA methylation in mammals remains unclear. Here, we reveal that WTAP is widely expressed from the early stage of tooth development. Specific inactivation of Wtap in mouse enamel epithelium by the Cre/loxp system leads to serious developmental defects in amelogenesis. In Wtap conditional KO mice, we determined that the differentiation of enamel epithelial cells into mature ameloblasts at the early stages of enamel development is affected. Mechanistically, loss of Wtap inhibits the expression of Sonic hedgehog (SHH), which plays an important role in the generation of ameloblasts from stem cells. Together, our findings provide new insights into the functional role of WTAP-mediated m6A methylation in amelogenesis in mammals.


What are the Top Research Priorities in Surgical Simulation and How Can They Be Best Addressed? Results From a Multidisciplinary Consensus Conference.
To define the top priorities in simulation-based surgical education where additional research would have the highest potential to advance the field and develop proposals that would address the identified research priorities. Simulation has become integral part of surgical training but there are a number of outstanding questions that have slowed advances in this field. The Delphi methodology was used to define the top priorities in simulation-based surgical education. A research summit was held with multiple stakeholders under the auspices of the American College of Surgeons Division of Education to develop proposals to address these priorities. Consensus was achieved after the first round of voting on the following 3 most important topics: (1) impact of simulation training on patient safety and outcomes, (2) the value proposition of simulation, and (3) the use of simulation for physician certification and credentialing. Knowledge gaps, challenges and opportunities, and research questions to address these topics were defined by summit participants. The top 3 priorities in surgical simulation research were defined and project outlines were developed for impactful projects on these topics. Successful completion of such projects is expected to advance the field of simulation-based surgical education.


Comprehensive analyses of genome-wide methylation and RNA epigenetics identify prognostic biomarkers, regulating the tumor immune microenvironment in lung adenocarcinoma.
The aim of our study was to identify a signature of immune-regulated molecules and reveal its prognostic role in lung adenocarcinoma (LUAD). We downloaded RNA-Sequencing data and DNA methylation data from the Gene Expression Omnibus (GEO) database. GEO2R was used to analyze differentially expressed mRNAs (DEmRNAs). we used "factoextra" R package to do the principal component analysis (PCA) of DEmRNAs. "Limma" R package was used to identify DEmRNAs, differentially expressed miRNAs (DEmiRNAs), differentially expressed lncRNAs (DElncRNAs) from The Cancer Genome Atlas (TCGA) database. Three R packages "org.Hs.eg.db", "clusterProfiler", "ggplot2″ were used to show enrichment results. Considering about methylation and mutation data, TEK and SOX17 mediated cancer signaling pathways. Through tumor-immune system interactions database (TISIDB) and Tumor Immune Estimation Resource (TIMER), higher methylated and lower expressed TEK may act as a prognostic marker, regulating the tumor immunity in LUAD. Through four databases (MEXPRESS, DNMIVD, MethSurv, Firehose), we further verified the methylation (P = 2.33e-23) and mutation about TEK. A signature of immune-associated TEK to predict survival of LUAD patients was validated. Prognostic, methylation, immune microenvironment analysis showed new light on potential novel therapeutic targets in LUAD.


Ordinary and opportunistic enteropathogens associated with diarrhea in Senegalese adults in relation to human immunodeficiency virus serostatus.
A survey was conducted in Dakar, Senegal, to identify major types and prevalences of bacteria, parasites, fungi, and Rotaviruses associated with diarrhea in relation to human immunodeficiency virus (HIV) serostatus with the goal to provide guidance to physicians for case management. Etiologic agents were identified in a case control study: cases were HIV-infected patients with diarrhea (HIV+ D+) and HIV seronegative patients with diarrhea (HIV D+); controls were HIV-infected patients without diarrhea (HIV+ D ) and seronegative controls without diarrhea (HID D ). Ordinary enteric pathogens were identified by conventional methods. Different Escherichia coli pathotypes were characterized by polymerase chain reaction (PCR), identification of HEp-2 cell adherence pattern, Sereny test, GM1-ELISA, and the suckling mouse assay. Opportunistic parasites, such as Cryptosporidium and Microsporidium, were identified by the Kinyoun method and trichromic stain of Weber, respectively. Rotaviruses were identified with a commercial latex agglutination kit. Antimicrobial susceptibility testing was carried out by the disk diffusion method. Among the 594 patients examined, 158 were HIV+ D+, 121 were HIV2 D+, 160 were HIV+ D , and 155 were HIV D . The main etiologies of diarrhea were different according to HIV serostatus of patients. In immunocompetent adults the main causes of diarrhea were Shigella sp (12.4%), Entamoeba histolytica(10.7%), Salmonella enterica (6.6%), and Giardia (4.9%). In the immunocompromised host the more frequent pathogens were enteroaggregative E. coli (19.6%), Microsporidium (9.4%), Cryptosporidium sp (8.2%), Rotavirus (8.2%), Shigella sp (7.6%), Candida albicans (7.6%), E. histolytica (5.1%), S. enterica (4.4%), and Isospora belli (4.4%). Also, Blastocystis hominis has to be considered as an opportunistic parasite, because it was identified only in HIV-infected patients, with higher prevalence in adults with diarrhea (2.5% in HIV+ D+ patients; 0.6% in HIV+ D patients). High level of asymptomatic carriage of Ascaris lumbricoides and Trichuris trichiura and some cases of multiple infections were observed. Fungi, Cryptosporidium sp and Microsporidium sp, were often identified in patients with low CD4 counts (range, 79 250 cells/mL). Independently from HIV-serostatus, CD4 count was lower in diarrheic persons, suggesting that diarrhea is a debilitating illness and that effective management of diarrhea can prevent immunosuppression. Isolated enteropathogenic strains displayed high resistance to most antibiotics used in Senegal for treating diarrhea (ampicillin, tetracycline, cotrimoxazole); they were susceptible to amikacin, gentamicin, and norfloxacin. These epidemiologic data suggest that guidelines for the management of diarrhea during HIV infection in Dakar should be updated.