Effect of Whitening Toothpaste on Surface Roughness and Colour Alteration of Artificially Extrinsic Stained Human Enamel: In Vitro Study.
Objective: The aim of this study was to compare five toothpastes in terms of alteration of surface roughness and colour of red-wine-stained human enamel over time after brushing simulation. Methods: Stained specimens were randomly divided into five groups (n = 8): Oral-B Gum and Enamel (C), ZACT Stain Fighter (ZW), Colgate Optic White Volcanic Mineral (CW), Oral-B 3D White Luxe Fresh Breath (3DW), and Thepthai (TW). The colour and surface roughness of the specimens were measured after brushing simulation for four durations; two weeks, one month, six months, and twelve months. Abrasive particles in toothpaste were inspected under a scanning electron microscope. Results: Surface roughness was increased in the specimens that underwent brushing simulation in all groups (p < 0.05). ZW (6.33 ± 0.98 μm) exerted the most changes in surface roughness for all durations. Other groups showed similar surface roughness at each time point. ΔE00 and ΔL were increased in all groups until twelve months; however, there were no significant differences between C (ΔE00 = 30.17 ± 4.16, ΔL = 38.19 ± 4.34), CW (ΔE00 = 24.25 ± 10.52, ΔL = 31.12 ± 11.00), and TW (ΔE00 = 29.00 ± 3.96, ΔL = 36.68 ± 4.53) at any time period. Moreover, ZW (ΔE00 = 45.6 ± 8.01, ΔL = 53.03 ± 6.71) and 3DW (ΔE00 = 43.6 ± 7.33, ΔL = 51.03 ± 6.47) exhibited a substantial change and were statistically differed from the other groups after four-weeks. Various characteristics of abrasive particles were inspected under SEM. Conclusion: All five toothpastes increased the surface roughness altered the colour of red wine-stained human enamel over time.


Illness perceptions and changes in lifestyle following a gynecological cancer diagnosis: A longitudinal analysis.
This study explores patterns of lifestyle change and whether more threatening illness perceptions are associated with lifestyle changes post-treatment for smoking, alcohol consumption and Body Mass Index (BMI) among gynecological cancer patients. In total, 395 cancer patients (N=221 endometrial; N=174 ovarian) were included in this secondary analysis of longitudinal data. Lifestyle outcomes were assessed through self-reported questionnaires after initial treatment and 6, 12, and 18months of follow-up. Illness perceptions were assessed with the Brief Illness Perception Questionnaire (BIPQ). Latent class growth curve analyses were conducted to identify patterns of lifestyle change and linear mixed models using between-subject and within-subject effects to explore the association between BIPQ items and alcohol consumption (glasses/week) and BMI (kg/m2). After initial treatment, 15% (N=57) of the patients smoked, 53% (N=203) drank alcohol, and 60% (N=236) were overweight or obese. Overall, smokers made no considerable changes, but one subgroup of low level smokers reported positive decline. A slight decrease was observed for alcohol consumption among low and moderate level alcohol drinker subgroups, whereas BMI remained stable among endometrial cancer patients and increased for ovarian cancer patients. Moreover, patients with lower trust in their treatment to cure the disease drank more alcohol (β=0.32 glasses/week [95% CI 0.09; 0.56]). Change in lifestyle after a gynecological cancer treatment is not self-evident. Moreover, more threatening illness perceptions were not related to a healthier lifestyle. This study underlines the need for lifestyle-promoting activities to facilitate lifestyle improvement among gynecological cancer patients.


Association of pyrogenic exotoxin genes with pharyngitis and rheumatic fever/rheumatic heart disease among Indian isolates of Streptococcus pyogenes.
To monitor the presence of various pyrogenic exotoxin genes in strains of Streptococcus pyogenes isolated in India. Isolates recovered from pharyngitis (52) and rheumatic fever (RF)/ rheumatic heart disease (RHD) (8) patients were analysed for the presence of toxin genes, speA, speB and speF, by PCR. The specificity of the products was confirmed by restriction enzyme digestion and Southern hybridization. Among the 60 isolates studied, the incidence of speA, speB and speF were 5(8.3%), 56(93.3%) and 53(88.3%), respectively. The expression of these genes was established in representative isolates by RT-PCR. Comparative analysis of frequency of the speA, speB and speF genes, among pharyngitis and RF/RHD associated isolates, showed higher incidence in RF/RHD (25%, 100%,100%) as compared to pharyngitis patients (5.8%, 92.3%, 86.5%), respectively. SIGNIFICANCE OF STUDY: The presence of the speA gene, which is usually associated with scarlet fever or toxic shock-like syndrome, within few Indian isolates may be indicative of new virulent strains circulating within the Indian community. High distribution of toxin genes among RF/RHD compared to pharyngitis isolates indicate their possible role in increased virulence.


Outcomes of a National Institute of Allergy and Infectious Diseases Workshop on understanding HIV-exposed but seronegative individuals.
The fascinating conundrum that some individuals who are exposed to HIV in ways that would make viral transmission highly likely, yet are able to remain uninfected, has been appreciated for many years. As early as the late 1980s, reports of such individuals began appearing in the HIV/AIDS literature. Despite the critical importance of understanding possible mechanisms of natural HIV resistance for developing effective prevention strategies, numerous obstacles have prevented this essential area of scientific exploration from moving forward. The Workshop held on July 8-9, 2010 and supported by the Office of AIDS Research (OAR), the National Institute of Allergy and Infectious Diseases (NIAID), and the National Institute on Drug Abuse (NIDA) at the NIH hosted 200 participants and utilized the expertise of 42 AIDS researchers as invited speakers, session chairs, and discussion leaders for presentations and breakout sessions in an attempt to remove some of those obstacles. Accomplishments of the participants included developing a consensus for a new general term for the field, HIV-exposed seronegative (HESN), while recognizing the necessity to identify and utilize secondary descriptive criteria such as exposure level, risk group, duration of seronegativity, or natural resistance. Three key questions for future research were also identified by the group: (1) What is different in HESN versus those who get infected? (2) What is the immune response in HESN and is it just a marker of exposure or a correlate of protection? (3) What are the HESN host factors that help HESN resist infection? This report briefly summarizes the presentations, and describes future directions for addressing these questions and challenges.


Mucosal stimulation activates secretomotor neurons via long myenteric pathways in guinea pig ileum.
This study examined whether mucosal stimulation activates long secretomotor neural reflexes and, if so, how they are organized. The submucosa of in vitro full thickness guinea pig ileal preparations was exposed in the distal portion and intracellular recordings were obtained from electrophysiologically identified secretomotor neurons. Axons in the intact mucosa of the oral segment were stimulated by a large bipolar stimulating electrode. In control preparations, a single stimulus pulse evoked a fast excitatory postsynaptic potential (EPSP) in 86% of neurons located 0.7-1.0 cm anal to the stimulus site. A stimulus train evoked multiple fast EPSPs, but slow EPSPs were not observed. To examine whether mucosal stimulation specifically activated mucosal sensory nerve terminals, the mucosa/submucosa was severed from the underlying layers and repositioned. In these preparations, fast EPSPs could not be elicited in 89% of cells. Superfusion with phorbol dibutyrate enhanced excitability of sensory neurons and pressure-pulse application of serotonin to the mucosa increased the fast EPSPs evoked by mucosal stimulation, providing further evidence that sensory neurons were involved. To determine whether these reflexes projected through the myenteric plexus, this plexus was surgically lesioned between the stimulus site and the impaled neuron. No fast EPSPs were recorded in these preparations following mucosal stimulation whereas lesioning the submucosal plexus had no effect. These results demonstrate that mucosal stimulation triggers a long myenteric pathway that activates submucosal secretomotor neurons. This pathway projects in parallel with motor and vasodilator reflexes, and this common pathway may enable coordination of intestinal secretion, blood flow, and motility.


First Report of Black Dot Caused by Colletotrichum coccodes on Potato in the Tibet Autonomous Region of China.
Potato black dot causing by Colletotrichum coccodes is a common disease in potato throughout the world, infecting underground stems, tubers, roots and foliage. Potato is becoming the third main food crop produced on ~16,000 ha annually in the Tibet Autonomous Region of China, situated on the world's highest plateau. However, the disease causing by C. coccodes has not been reported in this region. During the disease survey in the Bailang County of Tibet in August, 2020, some potato plants cv. "JiZhang 12" with chlorosis and wilting of foliage were observed. The incidence of affected plants was 20%. Necrotic lesions were also observed on the basal stems of the affected plants. Three affected plants were collected for pathogen isolation and three isolates were obtained for further investigation. The colonies on potato dextrose agar (PDA) were initially white, turning gradually black with age and producing abundant black sclerotia. Conidia were cylindrical, hyaline, aseptate, guttulate, with average size of 13.80 to 18.55×4.94 to 5.35 μm for the three isolates in which 30 conidia each were measured. Such characteristics are similar to C. coccodes (Lees and Hilton, 2003). Mycelial growth rate was 0.69 to 0.74 cm/day at 25 oC over the three isolates. The three isolates were confirmed to be C. coccodes by species-specific PCR using primer set of Cc1NF1/Cc2NR1 producing 350 bp amplicons in the internal transcribed spacer (ITS) region according to Cullen et al. (2002). The Cc1NF1/Cc2NR1 sequences were identical for three isolates, therefore one sequence from isolate BL_JZ_J1 was submitted to the GenBank with accession number OM368349. Additional genes were also sequencing including the actin (ACT), chitin synthase l (CHS1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and another larger ITS region were also amplified from genomic DNA using primers sets ACT512F/ACT783R, CHS-79F/CHS-354R (Carbone and Kohn 1999), GDF1/GDR1 (Templeton et al. 1992) and ITS1/ITS4 (Glass and Donaldson 1995), respectively. Sequences obtained for those four regions were 216 bp, 218bp, 235bp and 576 bp, respectively. Each region in the three isolates were also identical, therefore one sequence for each region was submitted to the GenBank with accession number of OM417059, OM417060, OM417061, and OM349570, respectively, which had 100% similarity with C. coccodes of MN336525 (ACT), KU821274 (CHS1), KU821397 (GAPDH) and KU821175 (ITS), respectively. Phylogenetic analyses based on the concatenated sequences of those four loci showed that the BL_JZ_J1 was close to C. coccodes, a reference isolate of CPOS1 with the accession numbers of GQ856787 (ACT), GQ856723 (CHS1), GQ856744 (GAPDH) and GQ485588 (ITS) (Yang et al. 2009). Pathogenicity was confirmed by inoculating a conidia suspension (100 µl of 105 conidia/ml) on three stems of 6-week-old potato plant cv. 'Favorita' with an artificial wound generated by sterile toothpick for each isolate. An equal volume of sterile water was injected on the wound of three stems as a noninoculated control. Brownish necrotic lesions were observed on all inoculated stems 3 days post-inoculation under natural conditions, whereas control stems remained symptomless. Reisolation of the fungus from all inoculated symptomatic plants confirmed Koch's Postulates. To our knowledge, this is the first report of C. coccodes in potato in Tibet Autonomous Region of China. The finding of black dot in this region has important management implications for the growers since the pathogen can survive for long periods in the field both on potato debris and in soil.


Extrathoracic Metastases in Pleural Mesothelioma.
Guidelines recommend obtaining a computed tomography scan of the chest for the staging of pleural mesothelioma and for assessing response to treatment. Consensus is lacking regarding the necessity of serial imaging of distant extrathoracic sites. In this study, we determined the prevalence of extrathoracic metastases in patients with pleural mesothelioma. We conducted a retrospective review of patients with pleural mesothelioma treated at Massachusetts General Hospital between 1999 and 2022 who were referred for extrathoracic imaging during their disease course. Imaging reports were reviewed to determine sites of metastasis and calculate the time to development of extrathoracic metastasis. Overall survival and prevalence of extrathoracic metastasis were compared for patients with epithelioid versus nonepithelioid mesothelioma. The study included 148 patients, 69 (47%) of whom had undergone cytoreductive surgery. Histologic types included epithelioid (n = 82, 55%), biphasic (n = 49, 33%), and sarcomatoid (n = 10, 7%) mesothelioma. The median overall survival for the cohort was 24.0 months, specifically 34.7 months and 16.7 months for patients with epithelioid and nonepithelioid tumors, respectively (p < 0.001). There were 65 (44%) patients who developed extrathoracic metastases, with a median time to extrathoracic metastasis of 11.5 months. The most common sites of involvement were extrathoracic nodes (22%), peritoneum (20%), bone (11%), and liver (11%). Of the 76 patients referred for brain imaging, seven (9%) had brain metastases. The frequency of extrathoracic metastasis was identical for epithelioid and nonepithelioid mesothelioma (44%). Overall survival was shorter for patients who developed extrathoracic metastases (hazard ratio 5.9, p < 0.001). Patients with pleural mesothelioma often develop extrathoracic metastases, providing a rationale for routinely obtaining imaging that encompasses sites outside of the thoracic cavity.


Improvement of thermal stability of Streptomyces cholesterol oxidase by random mutagenesis and a structural interpretation.
Random mutagenesis was used to enhance the thermal stability of Streptomyces cholesterol oxidase. Four thermostable mutants were isolated and the following amino acid substitutions were identified: Ser103 to Thr (mutant S103T), Val121 to Ala (mutant V121A), Arg135 to His (mutant R135H) and Val145 to Glu (mutant V145E). The wild-type and mutant enzymes were purified and characterized. The properties of mutants S103T, V121A and R135H were similar to those of the wild type but they showed improved thermal stability. When the V145E mutation was introduced, the thermal stability of the enzyme was markedly increased and the optimum pH was desirably changed to encompass a broad range from acid to alkali. Analysis of multiple mutants constructed by site-directed mutagenesis showed that all the mutations except that of R135H had an additive influence on the other mutations. These mutational effects are discussed in terms of a three-dimensional structural model of the enzyme constructed on the basis of homology modelling.


Rapid emergence of echinocandin resistance during Candida kefyr fungemia treatment with caspofungin.
Echinocandin drugs are widely used for the treatment of candidemia. Resistance is considered rare, and only a few cases of breakthrough candidiasis in patients receiving echinocandin have been reported worldwide. We report here for the first time a Candida kefyr isolate that acquired echinocandin resistance very rapidly after the initiation of caspofungin treatment for candidemia. We characterized the FKS gene mutation responsible for the resistance via the comparison of isolates sampled before and during treatment.


Citizen perspectives on the future of healthcare.
The Citizen's Reference Panel was created to engage the people of Ontario in a dialogue about the directions for the future sustainability of healthcare. The primary concerns raised during the course of the panellists' deliberations included the need to (1) create a much closer integration of the health system's many providers and institutions, (2) accelerate the deployment of e-health solutions across the health system and (3) continue to focus on improving access to care, especially primary care and expanded community care services to reduce the pressure on institutional care.


Multiple hydatid cyst disease revealed by an expansive intracranial process: A case report.
Hydatid disease is a real public health problem in developing countries. Cerebral localization is rare; it does not exceed 2%. It is more frequent in children and adolescents. It is often primary in children. In adults, it is less frequent and generally associated with other localizations. We report the case of a 60-year-old Tunisian patient with pulmonary, hepatic and cerebral hydatid cyst, revealed by neurological symptoms with an expansive intracranial process on brain imaging.


[Treatment of non-alcoholic fatty liver disease: an interpretation of the European clinical guidelines for obesity, type 2 diabetes, and digestive disease].
Although many Chinese and international guidelines offer recommendations for the treatment of nonalcoholic fatty liver disease (NAFLD), there are also confusions in clinical practice, including the differences in drug indications, drug selection, the basic principles of lifestyle intervention, and key points in individual regimens between Chinese and international guidelines. In addition, there are also differences in many aspects including medical model and medical insurance system between the East and the West. These differences are great challenges for clinical practice, and the hepatologists in China have to face the challenges of localization when following the guidelines for the treatment of NAFLD.


[Expression of osteopontin mRNA and its protein in colorectal cancer and liver metastatic tissues].
To investigate the expression of osteopontine (OPN) in colorectal cancer and liver metastatic tissues and the significance of OPN in tumorigenesis and liver metastasis of colorectal cancer. The expression of OPN mRNA in 44 samples of colorectal cancer tissues and adjacent normal tissues and 20 samples of liver metastatic tissues was examined by FQ-PCR, using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as internal standard. S-P immunohistochemical method was utilized to detect the OPN protein in colorectal cancer and liver metastasis tissues. On the principle that the C(T) value is in inverse proportion to the log value of original copy number of target sequence, OPN C(T)/GAPDH C(T) was used to evaluate the mRNA expression level in different tissues. The OPN mRNA levels in colorectal cancer tissues were 0.85 approximately 1.47 (kappa +/- s = 1.15 +/- 0.14 ), significantly higher than those in adjacent normal tissues (0.94 approximately 1.47,kappa +/- s =1.32 +/- 0.18) and lower than those in liver metastatic tissues (0.92 approximately 1.30,kappa +/- s =1.07 +/- 0.10) (t =5.81, P =0.00 and t = 2.12,P =0.038), Immunohistochemistry showed that OPN protein was localized in the cytoplasm of colorectal cancer cells, metastatic cancer cells, and normal hepatocytes around metastatic tissue, but not in the normal colorectal tissues around the primary cancer and normal hepatocytes in persons without colorectal cancer. The expression of OPN mRNA is the highest in liver metastatic tissues, moderate in primary colorectal cancer tissues, and lowest in adjacent normal tissues of the metastatic liver cancer. OPN protein is located in cytoplasm of colorectal cancer cells. OPN is a potential marker for liver metastasis of colorectal cancer.


Remote temperature monitoring in ocular tissue using confocal Raman spectroscopy.
We demonstrated the feasibility of Raman spectroscopy for remote temperature monitoring within the aqueous humor of the rabbit eye in vivo. Using a confocal Raman spectroscopy system, Raman spectra from 2580 to 3800 cm(-1) were recorded in HPLC-grade water and in the aqueous humor of the rabbit eye under in vivo and ex vivo conditions within a temperature range of 14-34 degrees C. The ratio between the integrated Raman intensities of two temperature dependent OH-vibrational regions (OH2/OH1) in the spectra of water showed high linear dependence on temperature both in pure water [0.0049(+/-1.2%)T+0.4522(+/-0.5%), R2=0.99, n=50, p<0.05], as well as in the rabbit aqueous humor [0.0036(+/-2.8%)T+0.4966(+/-0.6%), R2=0.98, n=162, p<0.05] with a high degree of reproducibility and sensitivity ( approximately 0.2-0.7 degrees C). Raman spectroscopy can be used for high resolution and remote monitoring of temperature in the aqueous humor under in vivo conditions.


Considerations on the malignancy of papillary meningioma. Clinico-pathological study of eight cases.
Eight papillary meningiomas, out of a collection of 750 cases of meningiomas, have been studied clinically and anatomopathologically. 2 had long survival without recurrences, 2 died shortly after operation, 4 had one or more recurrences with death. The 2 cases with long survival showed, at microscopical examination, few mitoses. On the contrary, in the six cases come to death the histological features were characterized by marked pleomorphism, focal necroses and frequent typical and atypical mitoses. In these cases, the correlation between "more malignant" histological aspects and "more aggressive" clinical behaviour is undoubtedly suggestive.


Dietary supplements during the cold season increase rumen microbial abundance and improve rumen epithelium development in Tibetan sheep.
Livestock on the Qinghai-Tibetan Plateau are faced with extreme harsh winters and are often in negative energy balance during this period. Dietary supplementation can improve growth performance of Tibetan sheep and, consequently, we hypothesized that it would also increase microbial abundance and rumen epithelium development. To test this hypothesis, we examined the effect of feed supplementation during the cold season on rumen microbes, fermentation, epithelium development, and absorptive capability in Tibetan sheep. Eighteen 1-yr-old ewes (BW = 29.4 ± 1.79, kg) were offered oat hay ad libitum for 60 d and divided randomly into three groups: 1) no supplement; control group (CON); 2) urea-molasses lick block supplement (BS); and 3) concentrate feed supplement (CS). The ADG of CS ewes (143.3, g/d) was greater (P < 0.05) than BS ewes (87.9, g/d), which was greater (P < 0.05) than CON ewes (44.5, g/d). Serum concentrations of GH, IGF-1, and IGF-2 in the CS and BS groups were greater than in the CON group (P < 0.05). Greater relative abundance of protozoa, Ruminococcus albus, Fibrobacter succinogenes, Streptococcus bovis, and Ruminobacter amylophilus was observed in the CS and BS groups than in the CON group (P < 0.05), and relative abundances of rumen fungi, Butyrivibrio fibrisolvens, and Prevotella ruminicola in the CS group were greater than in the BS and CON groups (P < 0.05). Ruminal total VFA, ammonia, and microbial protein concentrations in the CS and BS groups were greater than in the CON group (P < 0.05), and in the CS group were greater than in the BS group (P < 0.05). Ruminal papillae width and surface area in the CS and BS groups were greater than in the CON group (P < 0.05), while in the CS group were greater than in the BS group (P < 0.05). The mRNA expressions of IGFBP5, NHE1 (sodium/hydrogen antiporter, isoform 1), DRA (downregulated in adenoma), and Na+/K+-ATPase (sodium/potassium ATPase pump) in ruminal epithelium were greater in the CS and BS groups than in the CON group (P < 0.05), and in the CS group was greater than in the BS group (P < 0.05), while NHE3 (sodium/hydrogen antiporter, isoform 3), MCT1 (monocarboxylate transporter 1), and MCT4 (monocarboxylate transporter 4) mRNA expressions in the CS group were greater than in the BS and CON groups (P < 0.05). It was concluded that supplementing Tibetan sheep during the cold season increases rumen microbial abundance and improves fermentation parameters, rumen epithelium development, and absorptive capability.


Clinico-biological refinement of BCL11B-related disorder and identification of an episignature: A series of 20 unreported individuals.
BCL11B-related disorder (BCL11B-RD) arises from rare genetic variants within the BCL11B gene, resulting in a distinctive clinical spectrum encompassing syndromic neurodevelopmental disorder, with or without intellectual disability, associated with facial features and impaired immune function. This study presents an in-depth clinico-biological analysis of 20 newly reported individuals with BCL11B-RD, coupled with a characterization of genome-wide DNA methylation patterns of this genetic condition. Through an international collaboration, clinical and molecular data from 20 individuals were systematically gathered, and a comparative analysis was conducted between this series and existing literature. We further scrutinized peripheral blood DNA methylation profile of individuals with BCL11B-RD, contrasting them with healthy controls and other neurodevelopmental disorders marked by established episignature. Our findings unveil rarely documented clinical manifestations, notably including Rubinstein-Taybi-like facial features, craniosynostosis, and autoimmune disorders, all manifesting within the realm of BCL11B-RD. We refine the intricacies of T cell compartment alterations of BCL11B-RD, revealing decreased levels naive CD4+ T cells and recent thymic emigrants while concurrently observing an elevated proportion of effector-memory expressing CD45RA CD8+ T cells (TEMRA). Finally, a distinct DNA methylation episignature exclusive to BCL11B-RD is unveiled. This study serves to enrich our comprehension of the clinico-biological landscape of BCL11B-RD, potentially furnishing a more precise framework for diagnosis and follow-up of individuals carrying pathogenic BCL11B variant. Moreover, the identification of a unique DNA methylation episignature offers a valuable diagnosis tool for BCL11B-RD, thereby facilitating routine clinical practice by empowering physicians to reevaluate variants of uncertain significance within the BCL11B gene.


Magnetic alignment experiment of fine graphite-crystals dispersed in He gas oriented to study alignment of crystalline-axes of nano-sized non-magnetic particles.
The ensemble of nano-sized crystals is expected to attain additional physical properties when preferential alignments of certain crystal-axes are achieved by a magnetic field. The reduction of temperature T may realize alignment even if the mole number of the particle N and the diamagnetic anisotropy per mole (Deltachi)(DIA) are considerably small for the nano-sized diamagnetic oxides, since alignment proceeds by the balance between the energy of rotational Brownian motion and field-induced anisotropy energy. Alignment of various basic inorganic oxides such as gypsum, quartz, forsterite, KDP or calcite, having a size of 20 nm diameter, is expected to occur by a field intensity of approximately 50 T at T = 10 K; this intensity is presently available at a high magnetic-field laboratory. It is expected that the magnetic alignment of nano-sized particles can be observed by dispersing the particles in He gas, as achieved recently for micron-sized graphite crystals; a cryogenic liquid cannot be used as a dispersing medium. Measured (Deltachi)(DIA) values accumulated for basic inorganic-oxides are explained quantitatively by assuming that individual bonding-orbital composing the material possesses a constant amount of diamagnetic anisotropy; hence the majority of diamagnetic nano-sized insulators are expected to show magnetic alignment at finite field intensity.


Applying instructional design theories to bioinformatics education in microarray analysis and primer design workshops.
The need to support bioinformatics training has been widely recognized by scientists, industry, and government institutions. However, the discussion of instructional methods for teaching bioinformatics is only beginning. Here we report on a systematic attempt to design two bioinformatics workshops for graduate biology students on the basis of Gagne's Conditions of Learning instructional design theory. This theory, although first published in the early 1970s, is still fundamental in instructional design and instructional technology. First, top-level as well as prerequisite learning objectives for a microarray analysis workshop and a primer design workshop were defined. Then a hierarchy of objectives for each workshop was created. Hands-on tutorials were designed to meet these objectives. Finally, events of learning proposed by Gagne's theory were incorporated into the hands-on tutorials. The resultant manuals were tested on a small number of trainees, revised, and applied in 1-day bioinformatics workshops. Based on this experience and on observations made during the workshops, we conclude that Gagne's Conditions of Learning instructional design theory provides a useful framework for developing bioinformatics training, but may not be optimal as a method for teaching it.


[A minority of thymic lymphomas induced in the rat by a murine radioleukosis virus presents insertion in the vicinity of c-myc and a majority, a new nonviral polyadenylated RNA].
Thymic lymphomas were induced in rats either with the cell culture-propagated radiation leukemia virus complex, RadLV/VL3, or with a molecularly cloned isolate, RadLV/VL3 (T + L +). Four of thirty lymphomas, that were examined for rearrangements of the c-myc domain, displayed alterations in the vicinity of the c-myc gene, compatible with the idea of proviral integration. One of the tumours was investigated further, and was shown to contain a full length-proviral insert upstream of c-myc. Eight of nine lymphomas, that were investigated with respect to RNA expression, contained a novel polyadenylated RNA which could be detected with a molecular probe derived from the U5 portion of the retroviral long terminal repeat, but not with probes derived from the U3 portion or from the whole retroviral genome. These findings suggest that a RadLV/VL3 (T + L +) provirus can induce or activate RNA synthesis from c-myc by an enhancer mechanism, and from another cellular gene by a promotion mechanism.


Deep Learning-Enabled Clinically Applicable CT Planbox for Stroke With High Accuracy and Repeatability.
Computed tomography (CT) plays an essential role in classifying stroke, quantifying penumbra size and supporting stroke-relevant radiomics studies. However, it is difficult to acquire standard, accurate and repeatable images during follow-up. Therefore, we invented an intelligent CT to evaluate stroke during the entire follow-up. We deployed a region proposal network (RPN) and V-Net to endow traditional CT with intelligence. Specifically, facial detection was accomplished by identifying adjacent jaw positions through training and testing an RPN on 76,382 human faces using a preinstalled 2-dimensional camera; two regions of interest (ROIs) were segmented by V-Net on another training set with 295 subjects, and the moving distance of scanning couch was calculated based on a pre-generated calibration table. Multiple cohorts including 1,124 patients were used for performance validation under three clinical scenarios. Cranial Automatic Planbox Imaging Towards AmeLiorating neuroscience (CAPITAL)-CT was invented. RPN model had an error distance of 4.46 ± 0.02 pixels with a success rate of 98.7% in the training set and 100% with 2.23 ± 0.10 pixels in the testing set. V-Net-derived segmentation maintained a clinically tolerable distance error, within 3 mm on average, and all lines presented with a tolerable angle error, within 3° on average in all boundaries. Real-time, accurate, and repeatable automatic scanning was accomplished with and a lower radiation exposure dose (all P < 0.001). CAPITAL-CT generated standard and reproducible images that could simplify the work of radiologists, which would be of great help in the follow-up of stroke patients and in multifield research in neuroscience.


Thermal and Photoinduced Copper-Promoted C-Se Bond Formation: Synthesis of 2-Alkyl-1,2-benzisoselenazol-3(2H)-ones and Evaluation against Mycobacterium tuberculosis.
2-Alkyl-1,2-benzisoselenazol-3(2H)-ones, represented by ebselen (1a), are being studied intensively for a range of medicinal applications. We describe both a new thermal and photoinduced copper-mediated cross-coupling between potassium selenocyanate (KSeCN) and N-substituted ortho-halobenzamides to form 2-alkyl-1,2-benzisoselenazol-3(2H)-ones containing a C-Se-N bond. The copper ligand (1,10-phenanthroline) facilitates C-Se bond formation during heating via a mechanism that likely involves atom transfer (AT), whereas, in the absence of ligand, photoinduced activation likely proceeds through a single electron transfer (SET) mechanism. A library of 15 2-alkyl-1,2-benzisoselenazol-3(2H)-ones was prepared. One member of the library was azide-containing derivative 1j that was competent to undergo a strain-promoted azide-alkyne cycloaddition. The library was evaluated for inhibition of Mycobacterium tuberculosis (Mtb) growth and Mtb Antigen 85C (Mtb Ag85C) activity. Compound 1f was most potent with a minimal inhibitory concentration (MIC) of 12.5 μg/mL and an Mtb Ag85C apparent IC50 of 8.8 μM.


Risk of psychological difficulties among children raised by custodial grandparents.
Although custodial grandchildren are believed to be at greater risk of emotional and behavioral problems than children in general, scant research has examined this important public health issue. This study involves data from 733 custodial grandmothers participating in a study funded by the National Institute of Mental Health on custodial grandparenting and 9,878 caregivers from the 2001 National Health Interview Survey (NHIS) who completed the Strengths and Difficulties Questionnaire (SDQ) in reference to target children between ages four and 17. These two data sets were used to determine if custodial grandmothers report greater emotional and behavioral difficulties, as measured by the SDQ, for custodial grandchildren than do caregivers of children from the NHIS normative sample. Custodial grandchildren fared worse than children from the NHIS sample across all domains measured by the SDQ subscales, regardless of the child's gender and whether reporters were recruited by population-based or convenience sampling methods. Comparisons within the sample of 733 custodial grandmothers showed that higher levels of difficulties were reported when grandmothers were caring for boys, were recruited by convenience versus population-based sampling, and were white. Comparisons of the banded scores computed for each SDQ subscale suggested that custodial grandchildren have different cutoff points than children in the general population for a likely diagnosis of a psychiatric disorder. Similar to other children in kinship care arrangements, custodial grandchildren are reported by their caregivers to have higher levels of behavioral and emotional disturbances than children in the overall U.S. population.


The mechanisms of inhibition of frog endplate currents with homologous derivatives of the 1,1-dimethyl-3-oxybutyl phosphonic acid.
The mode of inhibition of endplate currents by four esters of 1,1-dimethyl-3-oxybutyl phosphonic acid with different lipophilicities and molecule lengths were estimated by mathematical modeling based on previous electrophysiological data supplemented by several experiments with rhythmic stimulation. The aim was to discriminate between their receptor and non-receptor effects. It was shown that all esters have a two-component mechanism of depression: inhibition of the receptor open channel and allosteric modulation of the receptor-channel complex. The ratio of both functional components depends on the length and lipophilicity of the esters. Short and less lipophilic esters mostly act as open channel inhibitors and the rate of inhibition substantially depends on the rate of stimulation, i. e. probability of the receptor-channel opening. As the length of the ester radicals and their lipophilicity increased, these compounds were more active as allosteric receptor inhibitors, probably hindering the function of nAChRs from the lipid annulus.


Pediatric eosinophilic esophagitis in Portugal.
Eosinophilic esophagitis (EoE) is an increasingly frequent diagnosis in our clinical practice, mainly in pediatric age. Allergic responses to food and aeroallergens have been increasingly implicated in the etiology of this disease. We describe a retrospective data analysis of pediatric EoE patients followed in our Immunoallergology Department. Of the 25 children (22 male, average 10.8 years), 88% had prior history of rhinoconjunctivitis, 76% asthma, 48% eczema and 36% food allergy. After evaluation, we identified in 76% and 92% of patients food and aeroallergen sensitization, respectively; 68% had simultaneously food and inhalant sensitization and 96% had at least one positive test to aeroallergens or food allergens. The first (44%) and the most frequent (56%) symptom was dysphagia. The time between symptoms onset and the EoE diagnosis averaged 18.6 ± 29.4 months. A multidisciplinary approach is needed for a correct evaluation, intervention and follow-up of these patients.


Interferon-alpha enhances the production of leukotriene B4 in murine peritoneal macrophages stimulated by opsonized zymosan.
Murine peritoneal macrophages pretreated with interferon (IFN)-alpha and then stimulated by opsonized zymosan produced two to three times more LTB4 than untreated macrophages. However, PGE2 production was not changed by IFN-alpha. Meanwhile, IFN-gamma did not affect the production of LTB4 and PGE2. From the results it is considered that IFN-alpha can modulate inflammation or host defence through the production of LTB4.


Measuring absolute expression with microarrays with a calibrated reference sample and an extended signal intensity range.
Gene expression ratios derived from spotted-glass microarray experiments have become invaluable to researchers by providing sensitive and comprehensive indicators of the molecular underpinnings of cell behaviors and states. However, several drawbacks to this form of data have been noted, including the inability to relate ratios to absolute expression levels or to compare experimental conditions not measured with the same control. In this study we demonstrate a method for overcoming these obstacles. First, instead of cohybridizing labeled experimental and control samples, we hybridize each sample against labeled oligos complementary to every microarray feature. Ratios between sample intensities and intensities of the oligo reference measure sample RNA levels on a scale that relates to their absolute abundance, instead of to the variable and unknown abundances of a cDNA reference. We demonstrate that results from this type of hybridization are accurate and retain absolute abundance information far better than conventional microarray ratios. Next, to ensure the accurate measurement of sample and oligo reference intensities, which may differ by several orders of magnitude, we use a linear regression algorithm, implemented in a freely available PERL script, to combine the linear ranges of multiple scans taken at different scanner sensitivity settings onto an extended linear scale. We discuss future applications of our method to measure RNA expression on the absolute scale of number of transcripts per cell from any organism for which oligo-based spotted-glass microarrays are available.


Right ventricular-vascular coupling in heart failure with preserved ejection fraction and pre- vs. post-capillary pulmonary hypertension.
Many patients with heart failure with preserved ejection fraction (HFpEF) develop post-capillary pulmonary hypertension (PH) due to increased left-sided filling pressures. However, a subset of patients develops combined post- and pre-capillary PH. We studied the value of echocardiographic right-sided characterization for the discrimination between pre- vs. post-capillary PH in HFpEF, using invasive haemodynamics as gold standard. 102 consecutive HFpEF patients with simultaneous right heart catheterization and echocardiography were identified. Patients were divided into: 'no PH', 'isolated post-capillary PH', and 'post- and pre-capillary PH'. Systolic pulmonary arterial pressure (SPAP), tricuspid valve annular plane systolic excursion (TAPSE), right ventricular-vascular coupling (TAPSE/SPAP), and VO2-max were assessed. Primary endpoint was all-cause mortality. A total of 97 patients were included: 22% no PH, 47% isolated post-capillary PH, and 31% post- and pre-capillary PH. Patients with post- and pre-capillary PH had more often diabetes mellitus (47 vs. 24%, P = 0.04), had more heart failure hospitalizations (57 vs. 26%, P = 0.007) and lower VO2-max (10 vs. 13 mL/min/kg, P = 0.008), compared with those with isolated post-capillary PH. Patients with post- and pre-capillary PH also had more reduced TAPSE (17 vs. 21 mm, P = 0.001) and TAPSE/SPAP (0.3 vs. 0.5, P < 0.001). TAPSE/SPAP ratio <0.36 had a good accuracy to identify patients with additional pre-capillary PH (C-statistic 0.86, sensitivity 86% and specificity 79%). TAPSE/SPAP ratio was associated with increased mortality (HR 2.51 [95% CI 1.25-5.01], P = 0.009). Abnormal right ventricular-vascular coupling identifies patients with HFpEF and additional pre-capillary PH, and predicts poor outcome in HFpEF.


Self-reported cancer family history is a useful tool for identification of individuals at risk of hereditary cancer predisposition syndrome at primary care centers in middle-income settings: a longitudinal study.
Analysis of cancer family history (CFH) offers a low-cost genetic tool to identify familial cancer predisposition. In middle-income settings, the scarcity of individual records and database-linked records hinders the assessment of self-reported CFH consistency as an indicator of familial cancer predisposition. We used self-reported CFH to identify those families at risk for hereditary cancer syndromes in community-based primary care centers of a low-income Brazilian area. We also evaluated the consistency of the information collected by reassessing CFH five years later. We interviewed 390 families and constructed their pedigrees for genetic cancer risk assessment. We found 125 families affected by cancer, 35.2% with moderate to high risk of familial susceptibility to cancer, a number that represents a relatively high prevalence of potential hereditary cancer syndromes in the overall study sample. Upon reassessment of CFH in 14/20 families that were previously identified as having at least one first-degree and one second-degree relative affected by cancer, and presented moderate to high risk for developing cancer, 90% of initial pedigrees were confirmed. These results demonstrate the reliability of self-reports as a means of early identification of healthy individuals at risk, encouraging the wider use of this method in low- and middle-income primary care settings.


Natural recovery of corals after severe disturbance.
Ecosystem recovery from human-induced disturbances, whether through natural processes or restoration, is occurring worldwide. Yet, recovery dynamics, and their implications for broader ecosystem management, remain unclear. We explored recovery dynamics using coral reefs as a case study. We tracked the fate of 809 individual coral recruits that settled after a severe bleaching event at Lizard Island, Great Barrier Reef. Recruited Acropora corals, first detected in 2020, grew to coral cover levels that were equivalent to global average coral cover within just 2 years. Furthermore, we found that just 11.5 Acropora recruits per square meter were sufficient to reach this cover within 2 years. However, wave exposure, growth form and colony density had a marked effect on recovery rates. Our results underscore the importance of considering natural recovery in management and restoration and highlight how lessons learnt from reef recovery can inform our understanding of recovery dynamics in high-diversity climate-disturbed ecosystems.


[Cytoreductive radical prostatectomy for prostate cancer with minimal osseous metastases: results of a first feasibility and case control study].
Androgen deprivation therapy (ADT) represents the standard treatment for patients with prostate cancer (PCA) and osseous metastases. We explored the role of cytoreductive radical prostatectomy in PCA with low volume skeletal metastases in terms of a feasibility study. A total of 23 patients with biopsy proven PCA, minimal osseous metastases (≤3 hot spots on bone scan), absence of visceral or extensive lymph node metastases and a decrease in prostate-specific antigen (PSA) to <1.0 ng/ml after neoadjuvant ADT were included in the feasibility study (group A). The control group (group B) consisted of 38 men with metastatic PCA who were treated by ADT alone. Surgery-related complications, time to castration resistance, symptom-free, cancer-specific and overall survival were analyzed using descriptive statistical analyses. The mean age was 61 years (range 42-69 years) and 64 years (47-83) in groups A and B, respectively, with similar patient characteristics in terms of initial PSA level, biopsy Gleason score, clinical stage and extent of metastatic disease. The median follow-up was 34.5 months (7-75 months) and 47 months (28-96 months) in groups A and B, respectively. Median time to castration resistance was 40 months (9-65 months) and 29 months (16-59 months) in groups A and B, respectively (p=0.04). Patients in group A experienced significantly better clinical symptom-free (38.6 versus 26.5 months, p=0.032) and cancer-specific survival rates (95.6% versus 84.2%, p=0.043) whereas the overall survival was similar. In group A none of the men underwent palliative surgical procedures for locally progressing PCA compared to 29% in group B. Cytoreductive radical prostatectomy is feasible in well-selected men with metastatic PCA who responded well to neoadjuvant ADT. These men have a long life expectancy and the risk of locally recurrent PCA and local complications are reduced. Cytoreductive radical prostatectomy might be a treatment option in the multimodal management of PCA with minimal osseous metastases.


Intervention for Anxiety and Problem Behavior in Children with Autism Spectrum Disorder and Intellectual Disability.
There is little research on the functional assessment and treatment of anxiety and related problem behavior in children with autism spectrum disorder (ASD), particularly those with intellectual and developmental disability (IDD). In a recent study, we evaluated a multimethod strategy for assessing anxiety in children with ASD and IDD (Am J Intellect Dev Disabil 118:419-434, 2013). In the present study, we developed treatments for the anxiety and associated problem behavior in these same children. A multiple baseline design was used to evaluate the effectiveness of a multicomponent intervention package, incorporating individualized strategies from Positive Behavior Support and Cognitive Behavioral Therapy. During intervention, all three participants showed substantial decreases in anxiety and problem behavior and significant increases in respiratory sinus arrhythmia in the situations that had previously been identified as anxiety-provoking.


Single-Step DNA Detection Assay Monitoring Dual-Color Light Scattering from Individual Metal Nanoparticle Aggregates.
Efficiently detecting DNA sequences within a limited time is vital for disease screening and public health monitoring. This calls for a new method that combines high sensitivity, fast read-out time, and easy manipulation of the sample, avoiding the extensive steps of DNA amplification, purification, or grafting to a surface. Here, we introduce photon cross-correlation spectroscopy as a new method for specific DNA sensing with high sensitivity in a single-step homogeneous solution phase. Our approach is based on confocal dual-color illumination and detection of the scattering intensities from individual silver nanoparticles and gold nanorods. In the absence of the target DNA, the nanoparticles move independently and their respective scattering signals are uncorrelated. In the presence of the target DNA, the probe-functionalized gold and silver nanoparticles assemble via DNA hybridization with the target, giving rise to temporal coincidence between the signals scattered by each nanoparticle. The degree of coincidence accurately quantifies the amount of target DNA. To demonstrate the efficiency of our technique, we detect a specific DNA sequence of sesame, an allergenic food ingredient, for a range of concentration from 5 pM to 1.5 nM with a limit of detection of 1 pM. Our method is sensitive and specific enough to detect single nucleotide deletion and mismatch. With the dual-color scattering signals being much brighter than fluorescence-based analogs, the analysis is fast, quantitative, and simple to operate, making it valuable for biosensing applications.


Effects of subchronic exposure to cadmium chloride on endocrine and metabolic functions in rainbow trout Oncorhynchus mykiss.
Adult and juvenile rainbow trout, Oncorhynchus mykiss, were exposed for 30 days to CdCl2 in water (10 and 25 micrograms Cd/L for adults, and 1 and 5 micrograms Cd/L for juveniles) to investigate effects of subchronic exposures to Cd on the hypothalamo-pituitary-interrenal and the hypothalamo-pituitary-thyroid axis, and on metabolic status. liver size and glycogen content and body mass gain were significantly reduced in the exposed adult fish and similar changes were detected in the juveniles. Plasma cortisol levels increased after exposure to the lower dose but decreased at the higher dose. Plasma T3 and T4 tended to decrease in the exposed adult fish. No significant differences in plasma glucose or cholesterol levels were detected between the controls and the exposed fish, plasma calcium levels were lower in the exposed fish. The results indicate that long-term exposures to Cd at sublethal doses have adverse effects on the physiological status of the fish and that these effects are mediated partly through alterations of endocrine function.


Investigation of the mechanism of action of pyrogallol-phloroglucinol transhydroxylase by using putative intermediates.
Pyrogallol-phloroglucinol transhydroxylase from Pelobacter acidigallici, a molybdopterin-containing enzyme, catalyzes a key reaction in the anaerobic degradation of aromatic compounds. In vitro, the enzymatic reaction requires 1,2,3,5-tetrahydroxybenzene as a cocatalyst and the transhydroxylation occurs without exchange with hydroxy groups from water. To test our previous proposal that the transfer of the hydroxy group occurs via 2,4,6,3',4',5'-hexahydroxydiphenyl ether as an intermediate, we synthesized this compound and investigated its properties. We also describe the synthesis and characterization of 3,4,5,3',4',5'-hexahydroxydiphenyl ether. Both compounds could substitute for the cocatalyst in vitro. This indicates that the diphenyl ethers can intrude into the active site and initiate the catalytic cycle. Recently, the X-ray crystal structure of the transhydroxylase (TH) was published16 and it supports the proposed mechanism of hydroxy-group transfer.


Generation, compression, and propagation of pulse trains in the nonlinear Schrödinger equation with distributed coefficients.
The generalized nonlinear Schrödinger model with distributed dispersion, nonlinearity, and gain or loss is considered and the explicit, analytical solutions describing the dynamics of bright solitons on a continuous-wave background are obtained in quadratures. Then, the generation, compression, and propagation of pulse trains are discussed in detail. The numerical results show that solitons can be compressed by choosing the appropriate control fiber system, and pulse trains generated by modulation instability can propagate undistorsted along fibers with distributed parameters by controlling appropriately the energy of each pulse in the pulse train.


[A case of bilateral spermatocele].
A case of bilateral spermatocele is reported. A 46-year-old man was admitted to our hospital complaining of bilateral scrotal swelling. We obtained a colorless, opalescent fluid which contained numerous spermatozoa. The fluid volume, pH and gravity obtained from right spermatocele were 85 ml, 6.8 and 1.005, respectively, and those obtained from left side were 40 ml, 6.8 and 1.006, respectively. Spermatocelectomy was done under lumbal anesthesia. Both spematocele were found near the body of the epididymis. The wall of spermatocele had on epithelial lining of cuboidal cells. Twenty two cases of spermatocele reported in Japan since 1951 are reviewed.


Evaluation of commercially available serologic diagnostic tests for chikungunya virus.
Chikungunya virus (CHIKV) is present or emerging in dengue virus-endemic areas. Infections caused by these viruses share some common signs/symptoms, but prognosis, patient care, and persistent symptoms differ. Thus, accurate diagnostic methods are essential for differentiating the infections. We evaluated 4 CHIKV serologic diagnostic tests, 2 of which showed poor sensitivity and specificity.


Platelet FcgammaRIIA receptor surface expression is increased in patients with ESRD and is associated with atherosclerotic cardiovascular events.
Accelerated atherogenesis is a prominent feature of end-stage renal disease (ESRD). Expression of platelet FcgammaRIIA immunoglobulin G receptor (FcgammaR) was measured in 2 populations: (1) 48 patients with ESRD with a mean age of 50.7 +/- 2.3 (SEM) years and (2) 48 healthy age- and sex-matched controls aged 53.6 +/- 1.6 years. Platelet FcgammaR expression was enhanced significantly in patients with ESRD (mean fluorescence intensity [MFI], 7.88 +/- 0.42 [SEM]; 95% confidence interval [CI], 7.03 to 8.72 versus 5.07 +/- 0.21; 95% CI, 4.64 to 5.49; P < 0.0001). In patients with ESRD, multivariate analysis showed that hemoglobin level and diastolic blood pressure were related inversely to FCgammaR expression (P = 0.01 and P = 0.03, respectively). Atherosclerotic cardiovascular events (ACVEs) were recorded prospectively for the ESRD cohort. A total of 22 ACVEs, defined as the occurrence of myocardial infarction (10 events), cerebrovascular accident (7 events), and/or a peripheral vascular event (5 events), were observed. In the group with greater receptor expression (MFI > 7.785; n = 24), 13 of 17 patients (76.5%) experienced at least 1 ACVE, whereas only 4 of 17 patients (23.5%) with low FcgammaR expression (MFI < 7.785; n = 24) experienced an event (P = 0.007). (1) Uremia is associated with enhanced platelet FcgammaR expression, and (2) patients with greater platelet FcgammaR expression are significantly more likely to experience ACVEs; however, (3) the small sample size is a limitation and our work therefore is a hypothesis-generating pilot study that remains to be confirmed.


Mass screening for gastric cancer by X-ray examination.
The characteristics of early gastric cancer detected by gastric mass survey were examined. The progress of the screening test using the indirect X-ray examination and the tendency toward reduction in mortality from stomach cancer were evaluated. In 1981 the number of examinees in the gastric mass surveys amounted to over four millions in all Japan. From the data of the gastric mass survey in Miyagi Prefecture the rate of detection of gastric cancer was approximately 0.18% and the ratio of early cancer to surgically treated gastric cancer was 59.4% for 1980-1982. Furthermore, the number of m cancer, small cancer and minute cancer cases detected by the survey has increased year by year. These results are due to various advanced screening tests or close examination techniques. From our long-term follow-up study it is clearly indicated that the rate of deaths from stomach cancer in the screened individuals was half of the rate in the unscreened individuals.


Protein-Glutaminase Engineering Based on Isothermal Compressibility Perturbation for Enhanced Modification of Soy Protein Isolate.
Protein-glutaminase plays a significant role in future food (e.g., plant-based meat) processing as a result of its ability to improve the solubility, foaming, emulsifying, and gel properties of plant-based proteins. However, poor stability, activity, high pressure, and high shear processing environments hinder its application. Therefore, we developed an application-oriented method isothermal compressibility perturbation engineering strategy to improve enzyme performance by simulating the high-pressure environment. The best variant with remarkable improvement in specific activity and half-time, N16M/Q21H/T113E, exhibited a 4.28-fold increase compared to the wild type in specific activity (117.18 units/mg) and a 1.23-fold increase in half-time (472 min), as one of the highest comprehensive performances ever reported. The solubility of the soy protein isolate deaminated by the N16M/Q21H/T113E mutant was 55.74% higher than that deaminated by the wild type, with a tinier particle size and coarser texture. Overall, this strategy has the potential to improve the functional performance of enzymes under complex food processing conditions.


Design for a lithographically patterned bioartificial endocrine pancreas.
Cell encapsulation provides a means to transplant therapeutic cells for a variety of diseases including diabetes. However, due to the large numbers of cells, approximately on the order of a billion, that need to be transplanted for human diabetes therapy, adequate mass transport of nutrients such as oxygen presents a major challenge. Proof-of-concept for the design of a bioartificial endocrine pancreas (BAEP) that is optimized to minimize hypoxia in a scalable and precise architecture is demonstrated using a combination of simulations and experiments. The BAEP is composed of an array of porous, lithographically patterned polyhedral capsules arrayed on a rolled-up alginate sheet. All the important structural variables such as the capsule dimensions, pore characteristics, and spacing can be precisely engineered and tuned. Further, all cells are encapsulated within a single device with a volume not much greater than the total volume of the encapsulated cells, and no cell within the device is located more than 200 μm from the surrounding medium that facilitates efficient mass transport with the surroundings. Compared with gel-based encapsulation methods, our approach offers unprecedented precision and tunability of structural parameters as well as the volume of the encapsulated cells and consequently the amount of secreted insulin. Our work highlights the utility of lithography and self-assembly in the fabrication of micro- and nanostructured three-dimensional structures that simulate the function of natural endocrine organs.


Human monoclonal antibodies against Ross River virus target epitopes within the E2 protein and protect against disease.
Ross River fever is a mosquito-transmitted viral disease that is endemic to Australia and the surrounding Pacific Islands. Ross River virus (RRV) belongs to the arthritogenic group of alphaviruses, which largely cause disease characterized by debilitating polyarthritis, rash, and fever. There is no specific treatment or licensed vaccine available, and the mechanisms of protective humoral immunity in humans are poorly understood. Here, we describe naturally occurring human mAbs specific to RRV, isolated from subjects with a prior natural infection. These mAbs potently neutralize RRV infectivity in cell culture and block infection through multiple mechanisms, including prevention of viral attachment, entry, and fusion. Some of the most potently neutralizing mAbs inhibited binding of RRV to Mxra8, a recently discovered alpahvirus receptor. Epitope mapping studies identified the A and B domains of the RRV E2 protein as the major antigenic sites for the human neutralizing antibody response. In experiments in mice, these mAbs were protective against cinical disease and reduced viral burden in multiple tissues, suggesting a potential therapeutic use for humans.


Decrease in bone density in young male guinea pigs fed high levels of ascorbic acid.
These experiments were conducted to investigate the effects of high oral intakes of ascorbic acid on bone metabolism in young guinea pigs and to inquire into the mechanism mediating these effects. Male guinea pigs 14 days old fed 8.7% of ascorbic acid for 6 weeks had decreased bone density and decreased urinary hydroxyproline compared to control animals fed 0.2% of ascorbic acid. To assess whether these changes were due to the acidity of the diet or to vitamin activity, salts of ascorbic acid, salts of erythorbic acid and free erythorbic acid were fed. There were no significant bone changes in any of these groups. The results strongly indicate that the acidity of ascorbic acid plays a part in the decrease in bone density but do not rule out that some other metabolic property specific to ascorbic acid may also be implicated.


Combined Use of Calcium-channel Blockers With Ombitasvir/Paritaprevir/Ritonavir Exacerbates Peripheral Edema in Elderly Japanese Patients.
The antiviral agent ritonavir is a substrate for cytochrome P450 3A4 (CYP3A4); therefore, concomitant use of CYP3A4-metabolising drugs might cause adverse reactions to this drug. We investigated the plasma level of calcium channel blockers (CCBs) as CYP3A4 substrates and peripheral edema as a potential adverse drug reaction possibly caused by the anti-hepatitis C virus (HCV) regimen of ombitasvir/paritaprevir/ritonavir (OPR) and CCBs. We enrolled Japanese patients prescribed OPR for HCV infection. Peripheral edema was graded according to the Common Terminology Criteria for Adverse Events ver. 4. Plasma samples were collected on days 0, 7, 14, 28, and 42 after antiviral treatment, at the trough level. Out of 52 patients, 64% experienced grade 1 or grade 2 peripheral edema, but not grade 3. Concomitant use of CCBs significantly increased the emergence of grade 2 edema (62%), compared with patients treated solely with OPR (48%). The use of OPR significantly increased the plasma concentration of amlodipine. Peripheral edema in patients treated with OPR and CCBs, although tolerable, should be closely monitored.


DRB4 dsRBD1 drives dsRNA recognition in Arabidopsis thaliana tasi/siRNA pathway.
In Arabidopsis thaliana, endogenous trans-acting and exogenous siRNA pathways are initiated by the interaction of DRB4 with trigger dsRNA. Further, DCL4:DRB4 complex cleaves the dsRNA into 21 bp siRNA. Understanding molecular determinants and mechanistic details of dsRNA recognition by DRB4 is vital for inducing long-term RNAi-mediated gene regulation in plants. Here, we present solution structures of individual and concatenated DRB4 dsRBDs and demonstrate modes of dsRNA binding by employing NMR, ITC and site-specific mutagenesis. While both dsRBDs adopt the canonical α-β-β-β-α fold, key structural differences and ms-μs dynamics located at the RNA binding region were observed for dsRBD1. These features favor dsRBD1 to orient itself and make stronger tripartite contact with dsRNA, a feature missing in dsRBD2. Additionally, the inter-domain orientation induced by the linker restricts the mobility of dsRBD2, resulting in the steric hindrance of α1 helix in dsRBD2, and leads in further reduction of its dsRNA binding activity. Our study deciphers functional roles of DRB4 domains by showing that dsRBD1 drives the tasiRNA/siRNA pathway. Furthermore, we identify a potential role of the C-terminal region of DRB4 in protein:protein interaction as it possesses six PxxP motifs, binds to Zn2+ and contains a small structural domain.


Sexuality and androgens in women with cyclical mood changes and pre-menstrual syndrome.
To study the relation between androgen levels and sexual interest in women with different kinds of pre-menstrual syndrome (PMS). Causal comparative study. Swedish university hospital outpatient clinic. Seventy women with cyclical mood changes. Pre-menstrual syndrome patients were divided into those with and those without preovulatory symptoms. In 37 women, early follicular phase blood samples were analyzed for androstenedione, testosterone, sex hormone-binding globulin (SHBG), progesterone and estradiol, using radioimmunoassay. The participants were divided into subgroups depending on whether the levels of androgens and SHBG were above or below the median. In 33 of them it was possible to compare the cyclicity in sexual parameters between these subgroups. Daily ratings of sexual parameters and hormonal analyses. Plasma testosterone was significantly lower and SHBG significantly higher in women with luteal phase symptoms compared with those with additional follicular phase symptoms. ANOVA showed significant cyclicity for all sexual parameters consistently. For the "sexual feelings" and "pleasant sexual thoughts" parameters, cyclicity was the same whether or not the hormonal levels were "high" or "low." The "Pure-PMS" group and the "pre-menstrual-exacerbation" groups differed in their androgen and SHBG levels. Women suffering from PMS with higher neuroticism Eysenck Personality Inventory scores or "low" levels of androgens and SHBG would be more likely to have a decreased sexual interest pre-menstrually than would women with a high level.


A comprehensive overview on utilizing electromagnetic fields in bone regenerative medicine.
Stem cells are one of the most important sources to develope a new strategy for repairing bone lesions through tissue engineering. Osteogenic differentiation of stem cells can be affected by various factors such as biological, chemical, physiological, and physical ones. The application of ELF-EMFs has been the subject of many research in bone tissue engineering and evidence suggests that this exogenous physical stimulus can promote osteogenic differentiation in several types of cells. The purpose of this paper is to review the current knowledge on the effects of EMFs on stem cells in bone tissue engineering studies. We recapitulated and analyzed 39 articles that were focused on the application of EMFs for bone tissue engineering purposes. We tabulated scattered information from these articles for easy use and tried to provide an overview of conducted research and identify the knowledge gaps in the field.


Barbiturate coma for intracranial hypertension: clinical observations.
To determine the neurologic outcome of patients with intracranial hypertension treated with barbiturate-induced coma. The records of 49 patients who were admitted to a 31-bed medicosurgical intensive care unit over a 5-year period in whom a barbiturate coma was induced to control intracranial hypertension were analyzed retrospectively. Analysis included assessment of the response to barbiturate coma and evaluation of the long-term neurologic outcome according to the Glasgow Outcome Scale (GOS). Intracranial hypertension was caused by head trauma in 28 patients and subarachnoid hemorrhage in 21 patients. Eight of the head trauma patients and 5 of the patients with subarachnoid hemorrhage survived their hospital stay. The survivors were younger than the nonsurvivors, and had a good neurologic status after 1 year (except for 2 patients who died 1 and 3 months after discharge, respectively). There was no significant difference in the Glasgow Coma Score (GCS) on admission between the survivors and the nonsurvivors. The long-term outcome at 1 year was markedly better in the patients who had experienced a subarachnoid hemorrhage than in the trauma patients.


ErbB1 receptor ligands attenuate the expression of synaptic scaffolding proteins, GRIP1 and SAP97, in developing neocortex.
Scaffolding proteins containing postsynaptic density-95/discs large/zone occludens-1 (PDZ) domains interact with synaptic receptors and cytoskeletal components and are therefore implicated in synaptic development and plasticity. Little is known, however, about what regulates the expression of PDZ proteins and how the levels of these proteins influence synaptic development. Here, we show that ligands for epidermal growth factor receptors (ErbB1) decrease a particular set of PDZ proteins and negatively influence synaptic formation or maturation. In short-term neocortical cultures, concentrations of epidermal growth factor and amphiregulin (2-9 pM) decreased the expression of glutamate receptor interacting protein 1 (GRIP1) and synapse-associated protein 97 kDa (SAP97) without affecting postsynaptic density-95 (PSD-95) levels and glial proliferation. In long-term cultures, epidermal growth factor treatment resulted in a decrease in the frequency of pan-PDZ-immunoreactive aggregates on dendritic processes. A similar activity on the same PDZ proteins was observed in the developing neocortex following epidermal growth factor administration to rat neonates. Immunoblotting revealed that administered epidermal growth factor from the periphery activated brain ErbB1 receptors and decreased GRIP1 and SAP97 protein levels in the neocortex. Laser-confocal imaging indicated that epidermal growth factor administration suppressed the formation of pan-PDZ-immunoreactive puncta and dispersed those structures in vivo as well. These findings revealed a novel negative activity of ErbB1 receptor ligands that attenuates the expression of the PDZ proteins and inhibits postsynaptic maturation in developing neocortex.


The impact of attrition on the transmission of HIV and drug resistance.
Attrition due to loss to follow-up or termination of antiretroviral therapy (ART) among HIV-infected patients in care may increase the risk of emergence and transmission of drug resistance (TDR), diminish benefit of treatment, and increase morbidity and mortality. Understanding the impact of attrition on the epidemic is essential to provide interventions for improving retention in care. We developed a comprehensive HIV transmission dynamics model by considering CD4 + cell count dependent diagnosis, treatment, and attrition involving TDR and acquired drug resistance. The model was calibrated by 11 groups HIV/AIDS surveillance data during 2008-2018 from Guangxi, China, and validated by the prevalence of TDR among diagnosed treatment-naive individuals. We aimed to investigate how attrition would affect the transmission of HIV and drug-resistance when expanding ART. In the base case with CD4 + cell count dependent per capita attrition rates 0.025∼0.15 and treatment rates 0.23∼0.42, we projected cumulative total new infections, new drug-resistant infections, and HIV-related deaths over 2022-2030 would be 145 391, 7637, and 51 965, respectively. Increasing treatment rates by 0.1∼0.2 can decrease the above total new infections (deaths) by 1.63∼2.93% (3.52∼6.16%). However, even 0.0114∼0.0220 (0.0352∼0.0695) increase in attrition rates would offset this benefit of decreasing infections (deaths). Increasing treatment rates (attrition rates) by 0.05∼0.1 would increase the above drug-resistant infections by 0.16∼0.30% (22.18∼41.15%). A minor increase in attrition can offset the benefit of treatment expansion and increase the transmission of HIV drug resistance. Reducing attrition rates for patients already in treatment may be as important as expanding treatment for untreated patients.


Radioimmunoassay for luteinizing hormone releasing hormone in plasma.
A sensitive and specific double antibody radioimmunoassay has been developed capable of measuring LH-RH in extracted human plasma. Thyrotropin releasing hormone, lysine vasopressin and most of LH-RH analogues did not appear to affect the assay. Hypothalamic extract and some of the LH-RH analogues produced displacement curves which were parallel to that obtained with the synthetic LH-RH. Sensitivity of the radioimmunoassay was about 3 pg per assay tube. The coefficient of variation of intraassay was 6.4%, while that of interassay was 9.6%. Exogenous LH-RH could be quantitatively extracted by acidic ethanol when varying amounts of synthetic LH-RH were added to plasma. Immunoreactivity of LH-RH was preserved in plasma until 2 hr in the cold and gradually reduced thereafter. The plasma levels in LH-RH were 20 pg/ml or less in normal adults and not detectable in children. The aged males over 60 yr and postmenopausal women showed a tendency to have higher levels of plasma LH-RH. Plasma LH-RH level was significantly higher in midcycle than in follicular and luteal stages. The disappearance rate of LH-RH from the circulation after intravenous injection could be represented as half times of 4-6 min. Between 0.2-0.4% of the injected dose was excreted into urine within 1 hr. These results indicate that the determination of LH-RH might be a useful tool for elucidating hypothalamic-pituitary-gonad interactions.


Immunochemical characterization of the hyaluronic acid-hyaluronectin interaction.
The interaction between hyaluronic acid (HA) and hyaluronectin (HN) was analyzed by gel chromatography and by the effects of HA on the immunological precipitation of HN. This interaction led to formation of larger molecules, as shown by gel permeation. No inhibition of immune precipitation occurred in liquid phase after addition of HA, but the precipitates in unstained gels were rendered transparent, giving the appearance of inhibition. However, after staining of the gels the precipitates appeared normal. Moreover, a non-linear decrease of the diffusion rate in antibody-containing gel was observed as a function of HA concentration at HA:HN weight ratios of 0.75 x 10(-3) and higher. A faster movement during electrophoresis, depending on the HA:HN ratio, suppressed the precipitation line when tested by electrosyneresis and produced an increase in migration distance when tested by Laurell's electroimmunoassay. These results show that in the immunochemical detection and quantitation of NH by these techniques consideration should always be given to the amount of HA in the samples.


Scaling function for surface width for free boundary conditions.
We study the restricted curvature model with both periodic and free boundary conditions and show that the scaling function of the surface width depends on the type of boundary conditions. When the free boundary condition is applied, the surface width shows a new dynamic scaling whose asymptotic behavior is different from the usual scaling behavior of the self-affine surfaces. We propose a generalized scaling function for the surface width for free boundary conditions and introduce a normalized surface width to clarify the origin of the superrough phenomena of the model.


[Outpatient Psychotherapy for Children and Adolescents with Intellectual Disability].
Outpatient Psychotherapy for Children and Adolescents with Intellectual Disability The psychiatric morbidity risk for people with intellectual disability is considerably increased. Psychotherapy is considered as one possible treatment. Among experts the caresituation is described as substantial insufficient. This study investigated the perspective of psychotherapists for children and adolescents in the German state North Rhine-Westphalia regarding the care situation of outpatient psychotherapy for children and adolescents with intellectual disability. The participants received a questionnaire (a) about the care situation, (b) how they consider their training to treat these patients and (c) the possibility for free answers. In this cross-section study 185 licensed psychotherapists for children and adolescents filled out a semi-standardized online-questionnaire. The data were analyzed descriptively; free answers were categorized according to Mayring. Additionally, group differences were tested. (a) The psychotherapeutic supply situation for children and adolescents with intellectual disability was rated as insufficient. Clients with learning disability or minor intellectual disability are mainly treated. This group of people benefits from psychotherapy. (b) The training for treating these clients was rated as inadequate. (c) Qualitative analysis refers that request for psychotherapy is seen as difficult; cognitive orientated treatments need modification. The psychotherapeutic care situation for children and adolescents with intellectual disability appears insufficient. This topic should be part of the training curriculum for psychotherapists.


Identification of a hemagglutinin-specific idiotype associated with reovirus recognition shared by lymphoid and neural cells.
A xenogeneic antiserum raised to antireovirus immunoglobulin was used to define an idiotypic determinant present on antibodies to reovirus type 3 hemagglutinin. The same idiotype was identified on nonimmune lymphoid cells and on neuronal cells that specifically bind the hemagglutinin of type 3 reovirus. This idiotypic determinant, called Id3, is shared by (a) a monoclonal antibody to the neutralization site of hemagglutinin from type 3 reovirus; (b) BALB/c serum antibodies to the hemagglutinin of reovirus type 3; (c) R1.1, a murine thymoma cell line that binds reovirus type 3; (d) primary cultures of murine neuronal cells. The presence of an idiotype shared by antihemagglutinin antibodies and by structures on nonlymphoid cells suggests a general relationship between disparate receptors that recognize a common determinant. Furthermore, this suggests a novel approach for the study of viral receptor interactions and for analysis of mechanisms of autoimmune responses.


Cancer costs and outcomes in the Finnish population 2004-2014.
The cost of cancer and outcomes of cancer care has been much debated, since cancer represents 3-6% of total healthcare costs. The objective of this study was to analyse the development of the costs and outcomes in Finland between 2004 and 2014. The national cost, episodes and outcomes data were obtained from the national register databases. Two hospitals were used to validate the costs of care. The outcome measures included relative survival rate, mortality, sick leave days per patient and number of new disability pensions. The total cost of cancer in 2014 was 927 million €. The real costs increased by 1.7% per year over the period studied, while the cost per new cancer patient decreased. The relative survival rate was enhanced by 7%, and the number of sick leave days and new disability pensions per cancer patient was reduced. The share occupied by cancer treatment in total healthcare costs decreased slightly from 3.7% to 3.6%, indicating that cancer care has not become more expensive compared to the treatment of other diseases. This is the first survey to analyse the change in actual cancer costs and outcomes in the population-level within a 10-year period. Since cancer care outcomes in Finland have been among the best in Europe, the progress in terms of the costs and the conversions in the cost distributions across categories are significant and valuable sources for international comparisons.


Interaction between fat- and muscle development in children and adolescents.
Background: Prevalence of obesity increased noticeably during the last decades. Little is known so far about the association between fat accumulation due to obesity and skeletal muscle mass. The aim of this study was to describe the association between fat mass and muscle mass after adjusting for relevant confounding factors in the National Health and Nutrition Examination Survey (NHANES) study population of children and adolescents. We postulated a negative correlation between fat mass and lean body mass. Methodology: A total of 849 whole body DXA-scans of the NHANES study population of children and adolescents aged eight to twenty years of the years 1999-2004 were eligible for statistical analysis. Appendicular lean body mass (appLBM) was used to evaluate muscle mass. Bivariate analysis (Pearson's correlation coefficient), multiple linear regression analysis and mediation analysis were performed. The multiple regression analysis and mediation analysis were adjusted for weight, age height, sex ethnicity and physical activity. Results: Fat mass correlates with appendicular lean body mass (Pearons's r 0.336, p < 0.001). In the multiple linear regression analysis the regression coefficient between appLBM and FM was positive (0.204; p < 0.001), when considering appendicular lean body mass, fat mass and body weight, the regression coefficient was negative (-0.517; p < 0.001). Conclusions: Study results indicate a negative association of fat mass and skeletal muscle mass in children and adolescents, when weight, age, height, sex ethnicity and physical activity are considered. Further investigations are needed to evaluate if there is a biochemical interaction between fat cells and muscle cells that could explain this effect.


Blood in, blood out: left ventricular pseudoaneurysm following mitral valve endocarditis.
A 76-year old male on warfarin due to atrial fibrillation was admitted with Staphylococcus aureus septicaemia. Echocardiography demonstrated mitral valve endocarditis, and shortly thereafter, he suffered an intracranial haemorrhage as a result of septic embolism. Four weeks later, cardiac magnetic resonance imaging revealed a newly formed pseudoaneurysm. A left ventricular pseudoaneurysm caused by infective endocarditis is very rare, but awareness of this unusual complication may allow surgery to prevent rupture.


Prosthetic rehabilitation for a patient treated for embryonal rhabdomyosarcoma.
A female patient, now aged 17 years, was diagnosed with rhabdomyosarcoma (RMS) in the right pterygopalatine fossa when she was 3 years old. The RMS was successfully treated by excision, but the subsequent radiation and polychemotherapy resulted in the complete anesthesia of the distribution area of the right trigeminal nerve and loss of vision in the right eye. The patient also experienced pain in the mandibular joints and masticatory muscles. Panoramic radiographs displayed a multiple agenesia of the permanent teeth and underdeveloped apices. Treatment involved the fabrication of a complete maxillary denture. A removable device was fabricated to evaluate her response to an occlusal vertical dimension increase of 6 mm and provide a stable intercuspal position. After wearing the prosthesis for 6 months, the patient reported that she was completely free of symptoms.


[Effects of bortezomib at different doses in combination with dexamethasone in treatment of relapsed or refractory multiple myeloma: a comparative study].
To investigate the efficacy and toxicity of bortezomib of different doses in combination with dexamethasone for the treatment of relapsed or refractory multiple myeloma (MM). 23 patients with relapsed or refractory MM, 10 males and 13 females, aged 65 (42-86), were randomized to 2 groups: low dose group receiving intravenous 1.0 mg/m2 bortezomib (n=10) on days 1, 4, 6, and 11 (twice weekly) combined with dexamethasone 20 mg/d on days 1-4, with 3 weeks as a course of treatment, and (20 standard dose group: receiving intravenous 1.3 mg/m2 bortezomib (n=13) combined with dexamethasone. The patients were followed up for 9.5 (3-15) months. The effect was determined using modified European Group for Blood and Marrow Transplantation (EBMT) criteria. The complete response (CR) + partial response (PR) rate of the 1.0 mg/m2 bortezomib group was 70.0%, not significantly different from that of the 1.3 mg/m2 bortezomib group (61.5%, P > 0.05). The relief rate (CR + near CR rate) of the 1.0 mg/m2 bortezomib group was 20.0% not significantly different from that of the 1.3 mg/m2 bortezomib group (38.5%, P > 0.05). Only 1 case of adverse event over the grade 3 of National Cancer Institute Common Terminology Criteria for Adverse Events occurred in the 1.3 mg/m2 bortezomib group. Five cases of infectious fever and 2 cases of treatment-associated death occurred in the 1.3 mg/m2 bortezomib group. Bortezomib at the dose of 1.3 mg/m2 is more effective in treatment of relapsed or refractory MM than that at the dose of 1.3 mg/m2 and has more side effects.


A 2D Titanium Carbide MXene Flexible Electrode for High-Efficiency Light-Emitting Diodes.
Although several transparent conducting materials such as carbon nanotubes, graphene, and conducting polymers have been intensively explored as flexible electrodes in optoelectronic devices, their insufficient electrical conductivity, low work function, and complicated electrode fabrication processes have limited their practical use. Herein, a 2D titanium carbide (Ti3 C2 ) MXene film with transparent conducting electrode (TCE) properties, including high electrical conductivity (≈11 670 S cm-1 ) and high work function (≈5.1 eV), which are achieved by combining a simple solution processing with modulation of surface composition, is described. A chemical neutralization strategy of a conducting-polymer hole-injection layer is used to prevent detrimental surface oxidation and resulting degradation of the electrode film. Use of the MXene electrode in an organic light-emitting diode leads to a current efficiency of ≈102.0 cd A-1 and an external quantum efficiency of ≈28.5% ph/el, which agree well with the theoretical maximum values from optical simulations. The results demonstrate the strong potential of MXene as a solution-processable electrode in optoelectronic devices and provide a guideline for use of MXenes as TCEs in low-cost flexible optoelectronic devices.


Identification, Synthesis, and Characterization of Novel Baricitinib Impurities.
Baricitinib is a novel active pharmaceutical ingredient used in the treatment of rheumatoid arthritis, and it acts as an inhibitor of Janus kinase. During the synthesis of baricitinib, three unknown impurities were identified in several batches between 0.10 and 0.15% using high-performance liquid chromatography. The unknown compounds were isolated and identified as N-((3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-5-oxotetrahydrofuran-3-yl)methyl)ethane sulfonamide (lactone impurity, BCL), 2-(3-(4-(7H-[4,7'-bipyrrolo[2,3-d]pyrimidin]-4'-yl)-1H-pyrazol-1-yl)-1-(ethylsulfonyl)azetidin-3-yl)acetonitrile (dimer impurity, BCD), and 2-(1-(ethylsulfonyl)-3-(4-(7-(hydroxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)azetidin-3-yl) acetonitrile (hydroxymethyl, BHM). These compounds were synthesized and confirmed against the isolated samples. The structures of all the three impurities were confirmed by extensive analysis of 1H NMR, 13C NMR, and mass spectrometry. The lactone impurity formation was explained by a plausible mechanism. The outcome of this study was very useful for scientists working in process as well as in formulation development. To synthesize highly pure baricitinib drug substance, these impurities can be used as reference standards due to their potential importance.


A cross sectional study of sex differences in self-medication practices among university students in Slovenia.
Self-medication patterns in adults depend on sex. Self-medication among students is very common, but little is known about the influence of sex. The aim of the study was to determine the incidence of self-medication college students and to determine the effect of sex on self-medication patterns. A web based incidence study conducted on a sample of Slovenian university students. The main outcome measures were percentages of male and female students reporting the use of self-medication in the past year. A majority of students (92.3%) reported the use of some sort of self-medication in the past year. Most female students (94.1%) and most male students (90.9%) reported the use of self-medication in the past year. The difference was not statistically significant. More female students than male ones (p < 0.05) acquired the drugs for self-medication in pharmacies, used OTC drugs, herbal teas, herbs, vitamins and minerals, remedies for muscle mass gain, antibiotics, benzodiazepines, antacids, acetylsalicylic acid, topical corticosteroids, and nasal decongestives only with the advice of physicians or pharmacists, and thought that increasing drug dosage can be dangerous, that in case of side effects physicians' help must be sought, that no drug can be used during pregnancy, and that self-treatment can mask the symptoms and signs of diseases so the physicians can overlook them easily. Sex appears to be important factor in self-medication patterns even in young adults, such as students. The physicians should actively seek the presence of self-medication in this population. Inappropriate or unsafe use should be properly addressed and managed.


A Bayesian approach for estimating age-adjusted rates for low-prevalence diseases over space and time.
Age-adjusted rates are frequently used by epidemiologists to compare disease incidence and mortality across populations. In small geographic regions, age-adjusted rates computed directly from the data are subject to considerable variability and are generally unreliable. Therefore, we desire an approach that accounts for the excessive number of zero counts in disease mapping datasets, which are naturally present for low-prevalence diseases and are further innated when stratifying by age group. Bayesian modeling approaches are naturally suited to employ spatial and temporal smoothing to produce more stable estimates of age-adjusted rates for small areas. We propose a Bayesian hierarchical spatio-temporal hurdle model for counts and demonstrate how age-adjusted rates can be estimated from the hurdle model. We perform a simulation study to evaluate the performance of the proposed model vs a traditional Poisson model on datasets with varying characteristics. The approach is illustrated using two applications to cancer mortality at the county level.


The use of imaging techniques in understanding lower urinary tract (dys)function.
The ability to store urine in the bladder and to void at an appropriate time depends on several complex mechanisms in the lower urinary tract (LUT) and its neural control. Normal LUT function requires coordination of the urinary bladder, urethra, pelvic floor, efferent and afferent neurons and specific spinal cord and brain areas. These structures can be visualised using different imaging modalities, such as ultrasound, X-ray and magnetic resonance imaging. The supraspinal neural control of the LUT can be studied using functional brain imaging. During the last two decades, the many technological improvements of these imaging techniques have increased our knowledge of voiding dysfunction. Here, we review the different imaging modalities of the LUT and its neural control and discuss their importance for diagnosing and understanding voiding dysfunction.


Purinergic-mediated inhibition of Na+-K+-ATPase in proximal tubule cells: elevated cytosolic Ca2+ is not required.
The involvement of cytosolic Ca2+ concentration ([Ca2+]i) as messenger for the regulation of Na+-K+-ATPase activity was investigated in a renal cell line recently developed by immortalization of early proximal tubule primary cultures from the Wistar-Kyoto rat strain. Na+-K+-ATPase was measured as short-circuit current (Isc) in intact monolayers after permeabilization of the apical plasma membrane with amphotericin B. With symmetrical solutions, Isc quantitatively reflects Na+-K+-ATPase activity as judged by ouabain inhibition and dependence on Na+ and K+. Extracellular ATP (50% effective concentration = 0.32 mM) on the apical side produced acute inhibition of Na+-K+-ATPase-generated Isc of up to 50%. The inhibition peaked within 1 min and lasted approximately 5 min. The potency order was ATP > ADP >> beta,gamma-methyleneadenosine 5'-triphosphate = UTP, consistent with a P2y receptor. Extracellular ATP also stimulated a transient increase in [Ca2+]i. This increase had a similar time course as the inhibition of ATPase and reached a peak change of approximately 120 nM. However, the elevation of [Ca2+]i is not required in the purinergic inhibition of the Na+-K+-ATPase, since, first, increases in [Ca2+]i produced with a Ca2+ ionophore (ionomycin) failed to mimic the purinergic inhibition and, second, 1,2-bis(2-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid, which abolished the [Ca2+]i elevation, failed to block the purinergic inhibition.


Rates of inclusion of teenagers and young adults in England into National Cancer Research Network clinical trials: report from the National Cancer Research Institute (NCRI) Teenage and Young Adult Clinical Studies Development Group.
Poor inclusion rates into clinical trials for teenagers and young adults (TYA; aged 13-24 years) have been assumed but not systematically investigated in England. We analysed accrual rates (AR) from 1 April 2005 up to 31 March 2007 to National Cancer Research Network (NCRN) Phase III trials for the commonest tumour types occurring in TYA and children: leukaemia, lymphoma, brain and central nervous system, bone sarcomas and male germ cell tumours. AR for 2005-2007 were 43.2% for patients aged 10-14 years, 25.2% for patients aged 15-19 years, and 13.1% for patients aged 20-24 years in the tumour types analysed. Compared with accrual from 1 April 2005 to 31 March 2006, AR between 1 April 2006 and 31 March 2007 increased for those aged 10-14 and 15-19 years, but fell for those aged 20-24 years. AR varied considerably among cancer types. Despite four trials being available, patients over 16 years with central nervous system tumours were not recruited. Rates of participation in clinical trials in England from 2005 to 2007 were much lower for TYA older than 15 years compared with children and younger teenagers. The variations in open trials, trial age eligibility criteria and extent of trial activation in treatment centres in part explain this observation. Other possible influences, such as difficulties associated with the consent of TYA require further evaluation. Closer dialogue between those involved in planning and running trials for children and for adults is necessary to improve trial availability and recruitment. Further research is required to identify trends in trial availability and accrual for those tumours constituting the remaining 26% of TYA cancers.


HLA-DR2 predicts susceptibility and disease chronicity in Irish sarcoidosis patients.
HLA-DR2 (15) and 14 (6) have been recently proposed as susceptibility alleles for the development of sarcoidosis and HLA-DR15 as a marker of poor outcome, but validation in other populations is necessary. Employing serological techniques, we HLA-typed 103 Irish sarcoidosis patients and 105 ethnically-matched healthy controls for class I A and B and II DR and DQ alleles. HLA-B5 (10% vs. 2%, p = 0.018) and DR2 (45% vs. 27%, p = 0.007) were positively associated and B15 (0% vs. 7%, p = 0.01) negatively associated with sarcoidosis compared to control subjects. Seventy-five patients were followed > 2 years and 47 (63%) had chronic and 28 (37%) non-chronic disease. HLA-DR2 (55% vs. 27%, p = 0.001) and DR11 (26% vs. 5%, p<0.0001) were significantly more frequent in chronic disease vs. controls, in contrast to HLA-DR3 (13% vs. 38%, p = 0.002), which had a significant negative association. HLA-B5 (11% vs. 2%, p = 0.029) and DR3 (64% vs. 38%, p = 0.005) were significantly more frequent in non-chronic disease. Of the 29 patients achieving spontaneous remission, 24 (83%) were HLA-DR3 -positive and DR3-positivity was associated with significantly greater carbon monoxide diffusion at follow-up compared to DR3-negative patients (90% vs. 82% predicted, p = 0.027). This study supports the role of HLA-DR2 (15) as both a susceptibility and poor prognostic marker in sarcoidosis and DR2 positive patients may particularly benefit from close follow-up and early treatment. In contrast, DR3 positive patients are at a much lesser risk of chronic disease. Studies for long-term treatment effects require stratification for HLA-DR2 and DR3 status.


High-aspect-ratio structure fabrication on (110)-oriented silicon surfaces using tribo-nanolithography.
This study investigated methods of fabricating high-aspect-ratio structures on a silicon surface using a combination of tribo-nanolithography and wet chemical etching. Tribo-nanolithography forms an amorphous phase on a single-crystal silicon surface that has an etch resistance against potassium hydroxide so that a protruding structure can be fabricated by wet chemical etching. To fabricate high-aspect-ratio structures, (110)-oriented silicon was used, and the effect of machining parameters on the structure shape was investigated. The results showed that a structure with vertical side walls could be fabricated by machining along the (112) direction. The etch resistance against potassium hydroxide depended on the normal load and number of repetitions, and the width and the maximum height were functions of these conditions. The aspect ratio of the structure increased with the etch time as long as the amorphous phase was maintained. A sub-micrometer-scale high-aspect-ratio structure was fabricated based on these results, demonstrating the possibility of using this simple and effective method.


Hidden Structural Modules in a Cooperative RNA Folding Transition.
Large-scale, cooperative rearrangements underlie the functions of RNA in RNA-protein machines and gene regulation. To understand how such rearrangements are orchestrated, we used high-throughput chemical footprinting to dissect a seemingly concerted rearrangement in P5abc RNA, a paradigm of RNA folding studies. With mutations that systematically disrupt or restore putative structural elements, we found that this transition reflects local folding of structural modules, with modest and incremental cooperativity that results in concerted behavior. First, two distant secondary structure changes are coupled through a bridging three-way junction and Mg2+-dependent tertiary structure. Second, long-range contacts are formed between modules, resulting in additional cooperativity. Given the sparseness of RNA tertiary contacts after secondary structure formation, we expect that modular folding and incremental cooperativity are generally important for specifying functional structures while also providing productive kinetic paths to these structures. Additionally, we expect our approach to be useful for uncovering modularity in other complex RNAs.