Optimized cyclin D1 immunoperoxidase staining in mantle cell lymphoma.
Mantle cell lymphoma (MCL) has a worse prognosis than MALT lymphoma (MALTL). Distinction between MCL and MALTL on purely morphologic grounds can be difficult. Cyclin D1 (PRAD1/bcl1) is overexpressed in MCL as a result of a t(11:14) gene rearrangement, which leads to overexpression of cyclin D1 mRNA and protein. The immunohistochemical detection of cyclin D1 in MCL has been reported by several authors to be highly specific with sensitivity ranging from 70%-100%, but diagnostic laboratories have reported difficulty in finding a reliable method for cyclin D1 immunostaining. The aim of this study was to evaluate and optimize a method for detection of cyclin D1 by paraffin section immunoperoxidase staining. Sections of routinely processed tissue from five MCL and one splenic marginal zone lymphoma (MZL) were immunostained using a mixture of two primary monoclonal antibodies and a standard avidin-streptavidin method. Antigen retrieval was performed using 1) steam heat in citrate buffer, 2) as in "1" followed by sonication for one minute, and 3) as in "2" followed by enzymatic digestion. All the above were repeated, with the additional use of catalyzed signal amplification (CSA). Later, sections of the same cases, plus three MALTL were immunostained as in "2". Steam heat antigen retrieval alone produced the best results. All MCL showed positive nuclear staining while the MZL and all MALTL were negative. Sonication did not enhance staining noticeably, whereas enzymatic digestion produced cytoplasmic staining. CSA increased background staining with no significant gain in nuclear stain intensity. We conclude that cyclin D1 immunostaining of formalin-fixed, paraffin-embedded tissue can be reliably achieved by heat induces antigen retrieval and a cocktail of two monoclonal antibodies.


Influence of an unnatural amino acid side chain on the conformational dynamics of peptides.
In this work, a non-natural amino acid, H-propargylglycine-OH (Pra), is chosen to examine the side-chain effect on the backbone conformation of small peptides. The conformations of two synthesized Pra-containing tripeptides, Ac-Pra-Pra-NH(2) (PPTP) and Ac-Pra-Ala-NH(2) (PATP), are examined by infrared (IR) spectroscopy in combination with molecular dynamics (MD) simulations and quantum chemical computations. By analyzing the joint distributions of backbone torsional angles, several significant conformations can be identified for the two tripeptides solvated in D(2)O. At room temperature, 44 % of PPTP exists in the α-α conformation and 33 % of PATP exists in the α-polyproline-II conformation. Larger structural inhomogeneity is seen in both cases by MD simulations at elevated temperatures. Thus even a small side chain, such as the propargyl group can significantly alter the peptide backbone conformations. The results suggest that there is no overwhelming conformational propensity of the Pra residue in short peptides. IR spectra simulated in the amide-I region using two different methods, reasonably reproduce the experimental IR spectra and their temperature dependence.


[Application of the Portuguese version of the abbreviated instrument of quality life WHOQOL-bref].
The need of short instruments to evaluate Quality of life determines World Health Organization Quality of Life Group (WHOQOL Group) to develop an abbreviated version of the WHOQOL-100, the WHOQOL-bref. The objective is to present the Brazilian field trial of the WHOQOL-bref. WHOQOL-bref is composed by 26 questions divided in four domains: physical, psychological, social relationships and environment. The evaliation instrument, BDI (beck depression inventory) and BHS (beck hopelessness scale) were used in a 300 subjects sample in Porto Alegre, South Brazil. The instrument showed a good performance concerning internal consistency, discriminant validity, criterion validity, concurrent validity and test-retest reliability. The intrument allies good psychometric performance and practicity for use which puts it as an interesting option to evaluate quality of life in Brazil.


A randomized, controlled trial comparing long-term cosmetic outcomes of traumatic pediatric lacerations repaired with absorbable plain gut versus nonabsorbable nylon sutures.
To show that the use of absorbable sutures in pediatric traumatic lacerations affords good long-term cosmesis and no increase in complications (infection, dehiscence rates, and need for surgical scar revision) when compared with wounds sutured with nonabsorbable sutures. This was a randomized clinical trial conducted in a pediatric emergency department. Patients 1-18 years of age who presented to the emergency department with lacerations < 12 hours old were recruited between January 1999 and December 2001. Exclusion criteria were the following: wounds that could be approximated by tissue adhesives, animal/human bites, gross contamination, puncture/crush wounds, wounds crossing joints, lacerations of tendon/nerve/cartilage, collagen vascular disease, immunodeficiency, diabetes mellitus, bleeding disorder, and scalp lacerations. Patients were randomized into one of two groups: those receiving absorbable plain gut sutures (group A) and those receiving nonabsorbable nylon sutures (group NA). Board-eligible/certified pediatric emergency physicians or clinical fellows performed laceration repair in a standardized approach. All wounds were reevaluated within ten days by a single research nurse who assessed the wounds using a previously validated wound evaluation score (WES) composed of six items (presence of step-off, contour irregularities, margin separation, edge inversion, extensive distortion, and overall cosmetic appearance). A score of 6/6 was considered optimal. The study nurse also noted the presence of infection and dehiscence. The patients were then seen by a single blinded plastic surgeon at four or five months who evaluated the wound using a previously validated visual analog scale of cosmesis (VAS). In addition, the surgeon repeated the WES and assessed the need for surgical scar revision. A total of 147 patients were eligible, and 52 patients declined to participate. Of the 95 patients enrolled, 50 were randomized to group A and 45 to group NA. The two groups had similar ages, gender distributions, rates of use of sedation or steri-strips, wound lengths and widths, mechanisms of injury, and wound locations. At the short-term follow-up, no difference was found in the proportion of optimal WES scores between group A (63% of patients) and group NA (49% of patients) (relative risk = 0.73; 95% confidence interval [95% CI] = 0.45 to 1.17). No difference was found in the rates of infection and dehiscence between the two groups. Sixty-three of the 95 patients presented for long-term follow-up. The groups remained similar with respect to patient and wound characteristics as well as wound location. The average VAS score at four months was 79 (95% CI = 73 to 85) for group A and 66 (95% CI = 55 to 76) for group NA. In addition, no differences were found in the proportion of optimal WES between group A (36% of patients) and group NA (28% of patients) at four months (relative risk = 0.88; 95% CI = 0.62 to 1.26). Surgical scar revision was recommended for only three patients, of whom two were in group A. No patients chose to have their scars revised. No differences were found between group A and group NA for the rates of dehiscence (2% vs. 11%; p = 0.07) and infection (0 vs. 2; p = 0.3). The use of plain catgut absorbable sutures in the repair of traumatic lacerations in children appears to be an acceptable alternative to nonabsorbable sutures because the long-term cosmetic outcome seems to be at least as good. In this study, plain gut suture material seemed to provide slightly better cosmesis. In addition, no difference was found in the rate of dehiscence or infection between the groups.


Clinical Manifestations of Polycystic Ovary Syndrome and Associations With the Vaginal Microbiome: A Cross-Sectional Based Exploratory Study.
Previous studies suggest that the vaginal microbiome is associated with polycystic ovary syndrome (PCOS). However, the clinical manifestations of PCOS are heterogeneous. Whether the vaginal microbiome is related with different clinical symptoms was unknown. In this cross-sectional study, 89 female patients with PCOS admitted to Zhongda Hospital (Nanjing, China) were included. Basic demographic information, health-related behaviors, clinical manifestations and sex hormone levels were comprehensively recorded for all patients. Vaginal swabs were acquired for microbiota sequencing of the V3-V4 region of the 16S rRNA gene. The prevalence of bacterial vaginitis and vulvovaginal candidiasis was 15.7% and 13.5%, respectively, within the PCOS patients, which were the most important factors affecting the vaginal microbiome (permutational multivariate analysis of variance test, R2 = 0.108, P = 0.001). The vaginal microbiome was associated with specific clinical manifestations of PCOS, including acanthosis nigricans, intermenstrual bleeding, pregnancy history, testosterone level and anti-müllerian hormone level, with P values < 0.05. The abundance of Lactobacillus crispatus was higher (P = 0.010) while that of Lactobacillus iners was lower (P = 0.036) among PCOS patients with elevated testosterone levels. Other potential bacterial biomarkers were not statistically significant after adjusting for confounding factors. No evidence of associations of other common manifestations of PCOS, such as obesity and acne, with the vaginal microbiome was obtained. Vaginal bacterial species among PCOS patients with variable clinical manifestations, especially differences in testosterone levels, are distinct. Further studies are essential to investigate the microbiota and molecular mechanisms underpinning this disease.


beta-Lactamase produced by a highly beta-lactam-resistant strain of Bacteroides fragilis: an obstacle to the chemotherapy of experimental mixed infections.
A strain of Bacteroides fragilis, which produces a metallo-beta-lactamase, was inoculated into pouches on the backs of rats together with a beta-lactamase-negative Escherichia coli highly sensitive to beta-lactam antibiotics. The mixed infection rat pouch model was treated with either flomoxef (susceptible to hydrolysis by the beta-lactamase produced by B. fragilis), or cefmetazole (relatively resistant to hydrolysis). In this model of mixed infection flomoxef showed weak in-vivo activity against E. coli, although showing the same strong activity in a model of single infection with E. coli. On the other hand, cefmetazole showed strong activity against E. coli, even in the model of mixed infection. The concentrations of both drugs in the pouches were decreased in infections with the strain of B. fragilis. There was a greater decrease in the concentration of flomoxef than of cefmetazole. Flomoxef was unstable whereas cefmetazole was relatively stable in the pouch exudates that had been infected with B. fragilis. These experimental data suggest that bacteria that produce a metallo-beta-lactamase decrease the in-vivo efficacy of beta-lactam antibiotics against other co-infecting bacteria. Thus, it is suggested that it is important in the chemotherapy of mixed bacterial infections that include these highly resistant beta-lactamase-producing bacteria to use antibiotics that are stable to hydrolysis by these enzymes.


Molecular diagnosis of Wilson disease using prevalent mutations and informative single-nucleotide polymorphism markers.
Wilson disease (WD) is an autosomal recessive disorder caused by defects in the ATPase, Cu(2+) transporting, beta-polypeptide gene (ATP7B) resulting in accumulation of copper in liver and brain. WD can be thwarted if detected at a presymptomatic stage, but occasional recombination during carrier detection with dinucleotide repeat markers flanking the WD locus may lead to faulty diagnosis. We examined the use of intragenic single-nucleotide polymorphism (SNP) markers to avoid this limitation. We prepared genomic DNA from the peripheral blood of Indian WD patients. By use of PCR, we amplified the exons and flanking regions of the WD gene and then performed sequencing to identify the nucleotide variants. We genotyped the SNPs in 1871 individuals by use of the Sequenom mass array system. We made linkage disequilibrium plots using Haploview software. We identified 1 mutation accounting for 11% (19 of 174) of WD chromosomes among patients in addition to 4 prevalent mutations characterized previously. Among 24 innocuous allelic variants identified, we selected 3 SNPs found to have high heterozygosity (>0.40) for the detection of mutant WD chromosomes. On analyzing these SNPs in 28 test individuals, who were sibs to 17 unrelated WD patients, we obtained unequivocal genotyping in 25 cases (approximately 89%). The remaining 3 cases were genotyped by dinucleotide repeat marker (D13S133). Sets of SNP markers are highly heterozygous across most world populations and could be used in combination with analysis of prevalent mutations as a comprehensive strategy for determining presymptomatic and carrier sibs of WD patients.


Psychiatric morbidity and referral rate among medical in-patients at Kenyatta National Hospital.
Psychiatric morbidity among 200 medical in-patients at Kenyatta National Hospital was determined by a two-stage screening procedure, using the Self Reporting Questionnaire (SRQ) and standardized psychiatric interview (SPI). 44 (22%) of the total sample of 200 patients who were interviewed met the pre-established criteria for psychiatric morbidity; of these 59% comprised affective disorders. 4 (9%) of the psychiatric morbidity cases were referred for psychiatric evaluation. Referral seemed to be related to severity of illness and a previous history of psychiatric illness. There was no evidence in the notes that the psychiatric problems had been detected, treated or dealt with in any other way by the medical team in 34 patients out of 44 with psychiatric morbidity. Increase in the mental health input in the training of all health workers with emphasis on recognition and management of some of the commoner psychological problems is recommended.


Study of fluxes at low concentrations of L-tri-iodothyronine with rat liver cells and their plasma-membrane vesicles. Evidence for the accumulation of the hormone against a gradient.
1. Influx and efflux of l-tri-[(125)I]iodothyronine with isolated rat liver parenchymal cells and their plasma-membrane vesicles were studied by a rapid centrifugation technique. 2. At 23 degrees C and in the concentration range that included the concentration of free l-tri-iodothyronine in rat plasma (3-5pm) influx into cells was saturable; an apparent K(t) value of 8.6+/-1.6pm was obtained. 3. At 5pm-l-tri-[(125)I]iodothyronine in the external medium the ratios of the concentrations inside to outside in cells and plasma-membrane vesicles were 38:1 and 366:1 respectively after 7s of incubation. At equilibrium (60s at 23 degrees C) uptake of l-tri-[(125)I]iodothyronine by cells was linear with the hormone concentration, whereas that by plasma-membrane vesicles exhibited an apparent saturation with a K(d) value of 6.1+/-1.3pm. 4. Efflux of l-tri-[(125)I]iodothyronine from cells equilibrated with the hormone (5-123pm) was constant up to 21 s; the amount that flowed out was 17.7+/-3.8% when cells were equilibrated with 5pm-hormone. When plasma-membrane vesicles were equilibrated with l-tri-[(125)I]iodothyronine (556-1226pm) 66.8+/-5.8% flowed out after 21 s. 5. From a consideration of the data on efflux from cells and binding of l-tri-[(125)I]iodothyronine to the liver homogenate, as studied by the charcoal-adsorption and equilibrium-dialysis methods, it appears that 18-22% of the hormone exists in the free form in the cell. 6. Vinblastine and colchicine diminished the uptake of l-tri-[(125)I]iodothyronine by cells but not by plasma-membrane vesicles; binding to the cytosol fraction was not affected. Phenylbutazone, 6-n-propyl-2-thiouracil, methimazole and corticosterone diminished the uptake by cells, plasma-membrane vesicles and binding to the cytosol fraction to different extents. 7. These results suggest that at low concentrations of l-tri-[(125)I]iodothyronine rat liver cells and their plasma-membrane vesicles accumulated the hormone against an apparent gradient by a membrane-mediated process. Contribution of cytoplasmic proteins to uptake by plasma-membrane vesicles was negligible. The amount of l-tri-[(125)I]iodothyronine required to achieve half-maximal uptake agrees with that occurring in the free form in the blood, conferring physiological importance to the transporting system in the plasma membrane of the liver cell.


[Role of medical information processing for quality assurance in obstetrics].
The paradigma of problem-orientated assuring of the professional quality of medical case is a kind of "control loop system" consisting of the following 5 steps: routine observation, identification of the problem, analysis of the problem, translation of problem solutions into daily practice and control as to whether the problem has been solved or eliminated. Medical data processing, which involves documentation, electronic data processing and statistics, can make substantial contributions especially to the steps of observation, identification of the problem, and follow-up control. Perinatal data collection, which has already been introduced in 6 Länder of the Federal Republic of Germany, has supplied ample proof of this. These operations were conducted under the heading "internal clinical assuring of quality with external aid". Those clinics who participated in this programme, were given the necessary aid in self-observation (questionnaires, clinical statistics), and they were also given comparative informative data to help them in identifying the problems (clinical profiles, etc.). It is entirely left to the responsibility of the clinics themselves--voluntary cooperation and guarantee of remaining anonymous being a matter of course -- to draw their own consequences from the collected data and to translate these into clinical everyday practice.


Natural orifice transluminal endoscopic surgery: The transvaginal route moving forward from cholecystectomy.
The advent of minimally invasive surgery and the advances in endoluminal flexible endoscopy have converged to generate a new concept in digestive surgery, whose acronym natural orifice transluminal endoscopic surgery (NOTES), has become a familiar term in the surgical community. NOTES has been performed through the mouth, the bladder, the rectum and the vagina. Of these four approaches, the vagina has gained most popularity for several reasons. It is not only readily accessible and easy to decontaminate but it also provides safe entry and simple closure. The transvaginal approach has been described in the experimental and the clinical setting as an option for cholecystectomy, nephrectomy, splenectomy, segmental gastrectomy, retroperitoneal exploration and bariatric surgery. However, larger series are needed to delineate the exact risks of this approach, and to transcend cultural barriers that impede its wider introduction. Prospective randomized trials will shed light on the definitive role of the vaginal approach in minimal invasive surgery of the future.


Noninvasive biomarkers for the diagnosis of steatohepatitis and advanced fibrosis in NAFLD.
Nonalcoholic fatty liver disease (NAFLD) is the most common cause of abnormal liver enzymes in both adults and children. NAFLD has a histologic spectrum ranging from simple steatosis to nonalcoholic steatohepatitis (NASH), advanced fibrosis, and cirrhosis. It is imperative to distinguish simple steatosis from NASH since the latter has a progressive disease course and can lead to end-stage liver disease. Liver biopsy has been considered as the gold standard for the diagnosis of NASH. However, liver biopsy is invasive, costly, and can rarely cause significant morbidity (risk of morbidity, 0.06-0.35%; risk of mortality, 0.1-0.01%). Imaging studies such as ultrasonography, computed tomography, and magnetic resonance imaging have limited sensitivity in detecting steatosis and cannot distinguish steatosis from NASH. Alanine aminotransferase (ALT) has been used as a surrogate marker for liver injuries. However, ALT is not an ideal marker for either diagnosis of NAFLD or distinguishing steatosis from NASH. Better noninvasive biomarkers or panels of biomarkers that are cheaper, reliable, and reproducible are urgently needed for patients with NASH to assist in establishing diagnosis, providing risk information, and monitoring disease progression and treatment response. In this article, we plan to concisely review the current advances in the use of biomarkers for the diagnosis of NASH.


Slow drying of a spray of nanoparticles dispersion. In situ SAXS investigation.
Nanoparticles confined in droplets of less than a picoliter are forced to organize in submicronic dry grains through solvent evaporation. The evolution of structures of the grains and the constituent nanoparticles during the slow drying process are investigated in situ with small-angle X-ray scattering (SAXS) for the first time. The scattering results have been explained on the basis of the equilibrium thermodynamics of the droplets in the drying tube. We demonstrate that this technique is really efficient in describing the internal arrangement of the nanoparticles inside the drying droplets. Distinction between an almost homogeneous repartition of the nanoparticles in droplets and formation of core shell like particles even in strongly polydispersed droplets can be made using SAXS.


Evaluation and optimization of preparative variables for controlled-release floatable microspheres prepared by poor solvent addition method.
The aim of this study was to evaluate and optimize preparative parameters for floatable theophylline microspheres prepared by the emulsion-solvent evaporation method. A three-factor three-level Box-Behnken design was employed using amount of poor solvent, temperature-increase rate and drug loading as independent factors, and percentage floating at 3 h and time required for 50% drug release as dependent variables. Simultaneous optimization of the parameters for maximum buoyancy and desirable drug release was conducted using a partitioned artificial neural network. A microsphere using 27.6% of drug loading, 0.29 degrees C/min of temperature-increase rate, and 1.7 mL of poor solvent was identified for maximizing buoyancy and sustaining drug release.


A model for empowerment of nursing in Iran.
While the Iranian nursing profession tries to reach to its full capacity for participating in the maintenance of public health, its desire to develop is strongly influenced by cultural, economic, and religious factors. The concept of empowerment is frequently used in nursing and the health services, particularly in relation to the quality of care, since the mission of nursing is to provide safe and quality nursing care thereby enabling patients to achieve their maximum level of wellness. When considering the importance of nursing services in any health system, the 54th World Health Assembly recommended that programs be designed to strengthen and promote the nursing profession. Since empowerment is crucial to the role of nurses, a qualitative study was conducted and aimed at designing a model for empowering nurses in Iran. A grounded theory approach was used for analyzing the participants' experiences, their perceptions and the strategies affecting empowerment. Data collection was done through Semi-structured interviews and participant observation. Forty-four participants were interviewed and 12 sessions of observation were carried out. Three main categories emerged from the data collected; these are "personal empowerment", "collective empowerment", and "the culture and structure of the organization." From the participants' perspective, empowerment is a dynamic process that results from mutual interaction between personal and collective traits of nurses as well as the culture and the structure of the organization. Impediments, such as power dynamics within the health care system hinder nurses from demonstrating that they possess the essential ingredients of empowerment. A model was designed for empowering the nursing profession in Iran. Implementing this model will not only define nursing roles, identify territories in the national healthcare system, but it will restructure nursing systems, sub-systems, and services. Currently no such model exists; therefore, restructuring of the nursing system, including its services, education and research subsystems is recommended.


Rapid signal transduction in living cells is a unique feature of mechanotransduction.
It is widely postulated that mechanotransduction is initiated at the local force-membrane interface by inducing local conformational changes of proteins, similar to soluble ligand-induced signal transduction. However, all published reports are limited in time scale to address this fundamental issue. Using a FRET-based cytosolic Src reporter in a living cell, we quantified changes of Src activities as a local stress via activated integrins was applied. The stress induced rapid (<0.3 s) activation of Src at remote cytoplasmic sites, which depends on the cytoskeletal prestress. In contrast, there was no Src activation within 12 s of soluble epidermal growth factor (EGF) stimulation. A 1.8-Pa stress over a focal adhesion activated Src to the same extent as 0.4 ng/ml EGF at long times (minutes), and the energy levels for mechanical stimulation and chemical stimulation were comparable. The effect of both stress and EGF was less than additive. Nanometer-scale cytoskeletal deformation analyses revealed that the strong activation sites of Src by stress colocalized with large deformation sites of microtubules, suggesting that microtubules are essential structures for transmitting stresses to activate cytoplasmic proteins. These results demonstrate that rapid signal transduction via the prestressed cytoskeleton is a unique feature of mechanotransduction.


External Validation of a Tube Thoracostomy Complication Classification System.
Tube thoracostomy (TT) is a commonly performed procedure which is associated with significant complication rates. Currently, there is no validated taxonomy to classify and compare TT complications across different populations. This study aims to validate such TT complication taxonomy in a cohort of South African trauma patients. Post hoc analysis of a prospectively collected trauma database from Pietermaritzburg Metropolitan Trauma Service (PMTS) in South Africa was performed for the period January 2010 to December 2013. Baseline demographics, mechanism of injury and complications were collected and categorized according to published classification protocols. All patients requiring bedside TT were included in the study. Patients who necessitated operatively placed or image-guided TT insertion were excluded. Summary and univariate analyses were performed. A total of 1010 patients underwent TT. The mean age was (±SD) of 26 ± 8 years. Unilateral TTs were inserted in n = 966 (96%) and bilateral in n = 44 (4%). Complications developed in 162 (16%) patients. Penetrating injury was associated with lower complication rate (11%) than blunt injury (26%), p = 0.0001. Higher complication rate was seen in TT placed by interns (17%) compared to TT placed by residents (7%), p = 0.0001. Complications were classified as: insertional (38%), positional (44%), removal (9%), infective/immunologic (9%), and instructional, educational or equipment related (0%). Despite being developed in the USA, this classification system is robust and was able to comprehensively assign and categorize all the complications of TT in this South African trauma cohort. A universal standardized definition and classification system permits equitable comparisons of complication rates. The use of this classification taxonomy may help develop strategies to improve TT placement techniques and reduce the complications associated with the procedure. V. Single Institution Retrospective review.


Formation of triads without the dihydropyridine receptor alpha subunits in cell lines from dysgenic skeletal muscle.
Muscular dysgenesis (mdg/mdg), a mutation of the skeletal muscle dihydropyridine receptor (DHPR) alpha 1 subunit, has served as a model to study the functions of the DHPR in excitation-contraction coupling and its role in triad formation. We have investigated the question of whether the lack of the DHPR in dysgenic skeletal muscle results in a failure of triad formation, using cell lines (GLT and NLT) derived from dysgenic (mdg/mdg) and normal (+/+) muscle, respectively. The lines were generated by transfection of myoblasts with a plasmid encoding a Large T antigen. Both cell lines express muscle-specific proteins and begin organization of sarcomeres as demonstrated by immunocytochemistry. Similar to primary cultures, dysgenic (GLT) myoblasts show a higher incidence of cell fusion than their normal counterparts (NLT). NLT myotubes develop spontaneous contractile activity, and fluorescent Ca2+ recordings show Ca2+ release in response to depolarization. In contrast, GLTs show neither spontaneous nor depolarization-induced Ca2+ transients, but do release Ca2+ from the sarcoplasmic reticulum (SR) in response to caffeine. Despite normal transverse tubule (T-tubule) formation, GLT myotubes lack the alpha 1 subunit of the skeletal muscle DHPR, and the alpha 2 subunit is mistargeted. Nevertheless, the ryanodine receptor (RyR) frequently develops its normal, clustered organization in the absence of both DHPR alpha subunits in the T-tubules. In EM, these RyR clusters correspond to T-tubule/SR junctions with regularly spaced feet. These findings provide conclusive evidence that interactions between the DHPR and RyR are not involved in the formation of triad junctions or in the normal organization of the RyR in the junctional SR.


Long-term incense use and the risk of end-stage renal disease among Chinese in Singapore: the Singapore Chinese health study.
Experimental studies have shown that exposure to incense burning may have deleterious effects on kidney function and architecture. However, the association between chronic exposure to incense smoke and risk of end-stage renal disease (ESRD) has not been reported in epidemiologic studies. We investigated this association in the Singapore Chinese Health Study, a prospective population-based cohort of 63,257 Chinese men and women of 45-74 years of age in Singapore during recruitment from 1993 to 1998. Information on the practice of incense burning at home, diet, lifestyle and medical history was collected at baseline interviews. ESRD cases were identified through linkage with the nationwide Singapore Renal Registry through 2015. We used Cox proportional hazards regression analysis to estimate hazard ratio (HR) and 95% confidence interval (CI) of ESRD associated with domestic incense burning. Among cohort participants, 76.9% were current incense users. After an average 17.5 years of follow-up, there were 1217 incident ESRD cases. Compared to never users, the multivariable-adjusted HR for ESRD risk was 1.05 (95% CI, 0.80 to 1.38) for former users and 1.26 (95% CI, 1.02 to1.57) for current users of incense. In analysis by daily or non-daily use and duration, the increased ESRD risk was observed in daily users who had used incense for > 20 years; HR was 1.25 (95% CI, 1.07 to 1.46). Conversely, the risk was not increased in those who did not use incense daily or who had used daily but for ≤20 years. Our findings demonstrate that long-term daily exposure to domestic incense burning could be associated with a higher risk of ESRD in the general population.


Antibacterial activity of new hydrophilic sealants: In vitro study.
Pits and fissures sealing and modulation of oral microbiota through probiotics are important preventive measures against dental decays. The aim of this study was to investigate the antibacterial activity of the Embrace™ WetBond™ Pit and Fissure Sealant (Pulpdent, USA) and UltraSeal XT® Hydro™ (Ultradent, USA) against selected oral bacteria and probiotics. The antibacterial effect of both sealants was tested both through planktonic growth inhibition test - 96-well microtiter plates and agar disk diffusion assay containing light-cured Embrace™ and UltraSeal XT® against Streptococcus mutans and two oral probiotics (Streptococcus salivarius and Lactobacillus reuteri). Embrace™ showed a stronger and broad activity against all the bacterial strains tested (P < 0.05) in planktonic growth inhibition test even at its lowest dose (10 μl), with inhibition rates higher than 90% in all cases. UltraSeal XT® Hydro™ showed a mild antibacterial activity against L. reuteri, with growth inhibition rates being 19% and 23% for 20 μl and 50 μl, respectively. Regarding agar disk diffusion test, both sealants showed exclusively an antibacterial activity by contact. According to these findings, it is recommended to carefully plan the timing for the administration of different preventive interventions, such as oral probiotics assumption and sealant application, to maximize their specific effectiveness. We suggest prescribing oral probiotics first and putting off the Embrace™ sealant application to the end of probiotic treatment. On the contrary, it is possible to administer L. reuteri simultaneously with the application of UltraSeal XT® since it elicits a minimal antibacterial action against this oral probiotic.


The location of the inferior angle of the scapula in relation to the spine in the upright position: a systematic review of the literature and meta-analysis.
Practitioners in several of the health care professions use anatomical landmarks to identify spinal levels, both in order to enhance diagnostic accuracy and to specifically target the site of intervention. Authoritative sources usually state the upright inferior scapular angle (IAS) aligns with the spinous process (SP) of T7, but some specify the T7-8 interspace or the T8 SP. The primary goals of this study were to systematically review the relevant literature; and conduct a meta-analysis of the pooled data from retrieved studies to increase their statistical power. Electronic searching retrieved primary studies relating the IAS to a spinal level, as determined by an imaging reference standard, using combinations of these search terms: scapula, location, landmark, spinous process, thoracic vertebrae, vertebral level, palpation, and spine. Only primary studies were included; review articles and reliability studies related to scapular position but lacking spinal correlations were excluded. Eight-hundred and eighty (880) articles of interest were identified, 43 abstracts were read, 22 full text articles were inspected, and 5 survived the final cut. Each article (with one exception) was rated for quality using the QUADAS instrument. Pooling data from 5 studies resulted in normal distribution in which the upright IAS on average aligns closely with the T8 SP, range T4-T11. Since on average the IAS most closely identifies the T8 SP in the upright position, it is very likely that health professionals, both manual therapists and others, who have been diagnosing and treating patients based on the IAS = T7 SP rule (the conventional wisdom), have not been as segmentally accurate as they may have supposed. They either addressed non-intended levels, or made numeration errors in their charting. There is evidence that using the IAS is less preferred than using the vertebra prominens, and may be less preferred than using the iliac crest for identifying spinal levels Manual therapists, acupuncturists, anesthesiologists, nurses, and surgeons should reconsider their procedures for identifying spinal sites in light of this modified information. Inaccurate landmark benchmark rules will add to patient variation and examiner errors in producing spine care targeting errors, and confound research on the importance of specificity in treating spinal levels.


B cells from patients with Graves' disease aberrantly express the IGF-1 receptor: implications for disease pathogenesis.
Graves' disease (GD) is an autoimmune process involving the thyroid and connective tissues in the orbit and pretibial skin. Activating anti-thyrotropin receptor Abs are responsible for hyperthyroidism in GD. However, neither these autoAbs nor the receptor they are directed against have been convincingly implicated in the connective tissue manifestations. Insulin-like growth factor-1 receptor (IGF-1R)-bearing fibroblasts overpopulate connective tissues in GD and when ligated with IgGs from these patients, express the T cell chemoattractants, IL-16, and RANTES. Disproportionately large fractions of peripheral blood T cells also express IGF-1R in patients with GD and may account, at least in part, for expansion of IGF-1R(+) memory T cells. We now report a similarly skewed B cell population exhibiting the IGF-1R(+) phenotype from the blood, orbit, and bone marrow of patients with GD. This expression profile exhibits durability in culture and is maintained or increased with CpG activation. Moreover, IGF-1R(+) B cells produce pathogenic Abs against the thyrotropin receptor. In lymphocytes from patients with GD, IGF-1 enhanced IgG production (p < 0.05) and increased B cell expansion (p < 0.02) in vitro while those from control donors failed to respond. These findings suggest a potentially important role for IGF-1R display by B lymphocytes in patients with GD in supporting their expansion and abnormal Ig production.


Substituent effects and chemoselectivity of the intramolecular Buchner reaction of diazoacetamide derivatives catalyzed by the di-Rh(ii)-complex.
A density functional theory (DFT) study was performed to reveal that the substituent effects in the α-site have an effect on the chemoselectivity of the intramolecular Buchner reaction of diazoacetamide catalyzed by Rh2(OAc)4. The substituent effect is investigated considering five different groups (Z = -Me, -OMe, -H, -CN and -C(O)Me) in the substrates. The substituent group in the α-site changes the electronegativity of the C-atom in carbene and affects the chemoselectivity. The basis of chemoselectivity is the distribution of products that was analyzed by DFT calculations. The barrier energy of the favorable pathway is clearly lower than that of the other pathways. Nucleophilic substituent groups, such as -H, -OMe and -Me, are regarded as electron-donating groups, which increase the electropositivity of the C-atom in carbene compounds and improve the reactivity of the aromatic addition reaction. Electrophilic substituent groups, such as -CN and -C(O)Me, are regarded as electron-withdrawing groups, which decrease the electropositivity of the C-atom in carbene compounds and favor the C-H activation step. The computational results showed that the main product is cycloheptatriene when Z = -H/-OMe. The main product is β-lactam when the substituent group is -CN/-C(O)Me. When the substituent group is -Me, the products are a mixture of γ-lactams, β-lactams and cycloheptatriene.


Charge-Segregated Stacking Structure with Anisotropic Electric Conductivity in NIR-Absorbing and Emitting Positively Charged π-Electronic Systems.
Squarylium-based π-electronic cation with an augmented dipole was synthesized by methylation of zwitterionic squarylium. The cation formed various ion pairs in combination with anions, and the ion pairs exhibited distinct photophysical properties in the dispersed state, ascribed to the formation of J- and H-aggregates. The ion pairs provided solid-state assemblies based on cation stacking. It is noteworthy that complete segregation of cations and anions was observed in a pseudo-polymorph of the ion pair with pentacyanocyclopentadienide as a π-electronic anion. In the crystalline state, the ion pairs exhibited photophysical properties and electric conductivity derived from cation stacking. In particular, the charge-segregated ion-pairing assembly induces an electric conductive pathway along the stacking axis. The charge-segregated mode and fascinating properties were derived from the reduced electrostatic repulsion between adjacent π-electronic cations via dipole-dipole interactions.


Differences between cytoplasmic surfaces of deep-etched heart and liver gap junctions.
We have compared the ultrastructures of the cytoplasmic surfaces (CS) of isolated, glutaraldehyde-fixed gap junctional pellets from rat ventricles and liver by rapid freezing on a liquid helium-cooled surface, freeze fracture, deep etching, and double-axis rotary replication (J. Microsc. Oxford 137: 121-123, 1984). Deep-etched unproteolyzed cardiac junctions [protein subunit relative molecular wt (Mr) 44,000-47,000], isolated with phenylmethylsulfonylfluoride (PMSF) [Am. J. Physiol. 246 (Heart Circ. Physiol. 15): H865-H875, 1984; C.K. Manjunath, G.E. Goings, and E. Page. Proteolysis of cardiac gap junctions during their isolation from rat hearts. J. Membr. Biol. In press.] had particulate CS, while proteolyzed cardiac junctions (subunit Mr 29,500) made without PMSF and liver junctions (Mr 28,000) made with or without PMSF had nonparticulate CS. Taken together with our previous findings that electron micrographs of thin-sectioned isolated unproteolyzed cardiac junctions have urea-resistant fuzzy CS coatings [Am. J. Physiol. 246 (Heart Circ. Physiol. 15): H865-H875, 1984], that proteolyzed cardiac junctions and isolated liver junctions lack this fuzzy layer, and that the CS of in situ cardiac junctions is fuzzy [J. Membr. Biol. 78: 147-155, 1984; Am. J. Physiol. 246 (Heart Circ. Physiol 15): H865-H875, 1984], and published data that in situ freeze-etched liver gap junctions are nonparticulate (Cell 30: 395-406, 1982), these new observations strongly indicate that CS components of heart and liver gap junction proteins are structurally different.


Surgical treatment of patients with lumbar spinal stenosis with associated scoliosis.
Spinal stenosis in combination with scoliosis frequently is seen in elderly patients. Patients typically present with a combination of symptoms attributable to neurogenic claudication and radicular pain, and symptoms of lower back pain. For patients in whom conservative treatment is not sufficient, surgical treatment can be done with careful consideration of the overall patient and his or her medical status. Surgical treatment is twofold; one purpose is to decompress the neural elements, the other purpose is to stabilize and realign the spine to as great a degree as possible. Appropriate balance of the spine at the end of the procedure is more important than the absolute amount of correction obtained. Stabilization and correction of the spine is done with pedicle screw-rod instrumentation and fusion, and the procedure must be done in an efficient and timely manner to involve the least amount of morbidity. There are two types of deformity typically seen, one is a degenerative lumbar scoliosis with no or minimal rotational deformity (Type I), and the other is a degenerative scoliosis often superimposed on a preexisting scoliosis with greater rotational deformity and greater loss of lordosis (Type II). Instrumentation and correction techniques differ for these two types of deformities, with shorter instrumentation procedures usually possible for the Type I deformity and longer instrumentation with sagittal plane reconstitution necessary for Type II deformity.


The effect of self-sorting and co-assembly on the mechanical properties of low molecular weight hydrogels.
Self-sorting in low molecular weight hydrogels can be achieved using a pH triggered approach. We show here that this method can be used to prepare gels with different types of mechanical properties. Cooperative, disruptive or orthogonal assembled systems can be produced. Gels with interesting behaviour can be also prepared, for example self-sorted gels where delayed switch-on of gelation occurs. By careful choice of gelator, co-assembled structures can also be generated, which leads to synergistic strengthening of the mechanical properties.


A Bayesian Network Approach for Friedreich Ataxia Severity Classification using Probability Modelling.
Friedreich ataxia (FRDA) requires an objective measure of severity to overcome the shortcoming of clinical scales when applied to trials for treatments. This is hindered due to the rarity of the disease resulting in small datasets. Further, the published quantitative measures for ataxia do not incorporate or underutilise expert knowledge. Bayesian Networks (BNs) provide a structure to adopt both subjective and objective measures to give a severity value while addressing these issues. The BN presented in this paper uses a hybrid learning approach, which utilises both subjective clinical assessments as well as instrumented measurements of disordered upper body movement of individuals with FRDA. The final model's estimates gave a 0.93 Pearson correlation with low error, 9.42 root mean square error and 7.17 mean absolute error. Predicting the clinical scales gave 94% accuracy for Upright Stability and Lower Limb Coordination and 67% accuracy for Functional Staging, Upper Limb Coordination and Activities of Daily Living.Clinical relevance- Due to the nature of rare diseases conventional machine learning is difficult. Most clinical trials only generate small datasets. This approach allows the combination of expert knowledge with instrumented measures to develop a clinical decision support system for the prediction of severity.


Development of the first axillary in vitro shoot multiplication protocol for coconut palms.
The coconut palm or "tree of life" is one of nature's most useful plants and the demand for its fruit is increasing. However, coconut production is threatened by ageing plantations, pests and diseases. Currently, the palm is exclusively propagated via seeds, limiting the amount of planting material. A novel micropropagation method is presented, based on axillary shoot formation. Apical meristems of in vitro coconut seedlings are cultured onto Y3 medium containing 1 µM TDZ. This induces the apical meristem to proliferate through axillary shoots in ~ 27% of the initiated explants. These axillary shoots are seen as white clumps of proliferating tissue and can be multiplied at a large scale or regenerated into rooted in vitro plantlets. This innovative micropropagation method will enable the production of disease-free, high quality in vitro plantlets, which will solve the worldwide scarcity of coconut planting material.


Clinical validation of qPCR Target Selector™ assays using highly specific switch-blockers for rare mutation detection.
The identification of actionable DNA mutations associated with a patient's tumour is critical for devising a targeted, personalised cancer treatment strategy. However, these molecular analyses are typically performed using tissue obtained via biopsy, which involves substantial risk and is often not feasible. In addition, biopsied tissue does not always reflect tumour heterogeneity, and sequential biopsies to track disease progression (eg, emergence of drug resistance mutations) are not well tolerated. To overcome these and other biopsy-associated limitations, we have developed non-invasive 'liquid biopsy' technologies to enable the molecular characterisation of a patient's cancer using peripheral blood samples. The Target Selector ctDNA platform uses a real-time PCR-based approach, coupled with DNA sequencing, to identify cancer-associated genetic mutations within circulating tumour DNA. This is accomplished via a patented blocking approach suppressing wild-type DNA amplification, while allowing specific amplification of mutant alleles. To promote the clinical uptake of liquid biopsy technologies, it is first critical to demonstrate concordance between results obtained via liquid and traditional biopsy procedures. Here, we focused on three genes frequently mutated in cancer: EGFR (Del19, L858, and T790), BRAF (V600) and KRAS (G12/G13). For each Target Selector assay, we demonstrated extremely high accuracy, sensitivity and specificity compared with results obtained from tissue biopsies. Overall, we found between 93% and 96% concordance to blinded tissue samples across 127 clinical assays. The switch-blocker technology reported here offers a highly effective method for non-invasively determining the molecular signatures of patients with cancer.


Analysis of ROR1 Protein Expression in Mice with Reconstituted Human Immune System Components.
Receptor tyrosine kinase-like orphan receptor 1 (ROR1) is an oncofetal antigen expressed on multiple tumors and has no significant expression on normal human tissues. ROR1 is highly upregulated in chronic lymphocytic leukemia (CLL) B cells. NOD-scid IL2rg-/- (NSG) mice engrafted with human CD34+ hematopoietic progenitor cells (huNSG) achieved multilineage human immune cell reconstitution including B cells, T cells, NK cells, and DCs. Like the CLL patients, huNSG mice have abnormally high percentage of CD5-expressing B cells in the periphery. In light of this, we aim to determine whether ROR1 is expressed on huNSG B cells. Using flow cytometry analysis, we found that ROR1 was highly expressed in a proportion of bone marrow, spleen, and blood B cells, which were mostly immature B cells. Transplantation of the oncogene TCL-1-transduced CD34+ cells in neonatal NSG mice did not increase the frequency of ROR1-expressing B cells, but the mouse with the highest engraftment of transduced cells developed a tumor-like lump consisting of a high percentage of ROR1-expressing B cells. This study highlights the potential use of huNSG mice to study B cell malignant diseases and to evaluate immunotherapeutics targeting ROR1.


Syntheses, structures, properties and DFT study of hybrid inorganic-organic architectures constructed from trinuclear lanthanide frameworks and Keggin-type polyoxometalates.
In this paper we report the synthesis and X-ray characterization of four novel hybrid inorganic-organic assemblies generated from H4SiW12O40 as Keggin-type polyoxometalates (POM) and, in three of them, a trinuclear lanthanide cluster of type {Na(H2O)3[Ln(HCAM)(H2O)3]3}(4+) is formed, where Ln metal is La in compound 1, Ce in compound 2, and Eu in compound 3 (H3CAM = chelidamic acid or 2,6-dicarboxy-4-hydroxypyridine). These compounds represent the first POM-based inorganic-organic assemblies using chelidamic acid as an organic ligand. The thermal stability of the organic ligand is crucial, since pyridine-2,6-bis(monothiocarboxylate) instead of chelidamic acid is used (compound 4) under the same synthesis conditions, the decomposition of the ligand to pyridine was observed leading to the formation of colorless crystals of a pseudo hybrid inorganic-organic assembly. In compound 4 the hybrid inorganic-organic assembly is not formed and the organic part simply consists of four molecules of protonated pyridine acting as counterions of the [SiW12O40](4-) counterpart. The luminescent properties of compounds and have been investigated and their solid state architectures have been analyzed. Whereas compound only shows ligand emission, the Eu(3+) emission in compound 3 is discussed in detail. We have found that unprecedented anion-π interactions between the POM, which is a tetra-anion, and the aromatic rings play a crucial role in the crystal packing formation. To the best of our knowledge, this is the first report that describes and analyzes this interaction in Keggin-type POM based inorganic-organic frameworks. The energetic features of these interactions in the solid state have been analyzed using DFT calculations in some model systems predicted by us.


Adverse effects of granulocyte donations.
Various reactions occur during cytapheresis by either intermittent or continuous flow centrifugation, because of the nature of the procedures themselves and because of the exposure of the donor to steroids, macromolecular agents, anticoagulants, and saline. Most reactions are vasovagal (commoner in intermittent flow procedures) and citrate-related (commoner in continuous flow). Only very rarely do these require the procedure to be stopped. Other less common reactions are discussed. In addition to actual reactions, blood component depletion may occur, especially with frequently repeated procedures in the same donor. Red cell, platelet, and plasma depletion can all force limitation of the frequency of cytapheresis. While most reactions are of little consequence, they can cause procedural failure or refusal of a donor to permit continuation of cytapheresis, and can interfere with donor recruitment.


Intravenous Procainamide for Predicting the Response of Sustained Ventricular Tachycardia to Type III Antiarrhythmic Drugs.
The use of serial electrophysiology studies to guide antiarrhythmic drug therapy in patients with ventricular tachycardia is both costly and time consuming. Intravenous procainamide administered during of the initial electrophysiology study has previously been shown to be useful in predicting the efficacy of oral antiarrhythmic medications (type I and III). The purpose of this study is to confirm that ventricular tachycardia suppression after intravenous procainamide correlates with suppression on oral class III antiarrhythmic medications (amiodarone and sotalol). This study included all patients with sustained ventricular tachycardia who underwent an initial electrophysiology study including an acute suppression trial with intravenous procainamide and a subsequent restudy on oral amiodarone or sotalol. The response to intravenous procainamide was then compared with these type III antiarrhythmic medications. Between January 1993 and May 1995, 360 patients underwent electrophysiology studies for suspected or documented ventricular arrhythmias. One hundred patients (28%) had an inducible sustained ventricular tachycardia, and 26 patients received both intravenous procainamide and subsequently oral amiodarone or sotalol. Acute infusion of procainamide provided a highly specific method for predicting suppression of oral amiodarone and sotalol (82% and 100% respectively). However, several patients who were not suppressed by intravenous procainamide were suppressed by oral sotalol resulting in lower overall predictive accuracy 12/15 (80%) for amiodarone vs. 5/11 (45%) for sotalol treated group. We conclude that the acute infusion of procainamide may help to predict ventricular tachycardia suppression after oral amiodarone and sotalol. A larger prospective trial is warranted to confirm this finding.


Electrodeposition of gallium in the presence of NH₄Cl in an ionic liquid: hints for GaN formation.
Group III-V semiconductors are important in the production of a variety of optoelectronic devices. At present, these semiconductors are synthesized by high vacuum techniques. Here we report on the electrochemical deposition of GaN which seems to form in quite a thin layer from NH4Cl and GaCl3 in an ionic liquid.


Hypomagnesaemia in chronic, stable asthmatics: prevalence, correlation with severity and hospitalization.
Magnesium (Mg) deficiency is associated with airway hyperreactivity, wheeze and impairment of lung function. The prevalence of hypomagnesaemia, and its effect on asthma is unclear. Therefore, this prospective study was undertaken to evaluate the prevalence of low serum Mg level, to correlate whether hypomagnesaemia (serum Mg level <0.74 mmol x L(-1)) is associated with severity of asthma and increased frequency of hospitalizations in chronic, stable asthmatics. Ninety-three chronic stable asthmatics, who were on regular follow-up in an asthma clinic, were studied. On attendance to the asthma clinic, each subject was interviewed and clinically evaluated. The serum Mg level (normal 0.74-1.2 mmol x L(-1)), as well as the severity of asthma was measured. The medications used were recorded. The total number of hospitalizations due to asthma exacerbation was scored for the last three months that preceded the date of measuring the serum Mg level. Twenty-five patients (27%) had a low Mg level with a mean+/-SD of 0.69+/-0.04 mmol x L(-1), while 68/93 (73%) had a normal Mg level (0.82+/-0.08 mmol x L(-1)). The number of hospitalizations in asthmatics with low Mg levels was 10/25 (40%), compared to 8/68 (12%) (p<0.01) in asthmatics with normal Mg levels. Multiple logistic regression analysis showed that severe asthma was the only factor associated significantly with hypomagnesaemia (p=0.04). No effect was observed for inhaled beta-agonist, inhaled steroid or theophylline therapy on serum Mg level. To conclude, hypomagnesaemia is common in chronic asthmatics. Chronic asthmatics with low Mg tend to have more hospitalizations than chronic asthmatics with normal Mg. Hypomagnesaemia was also associated with more severe asthma.


Perioperative microbiologic profile of the conjunctiva in photorefractive keratectomy.
Therapeutic soft contact lenses are used commonly as an adjunctive treatment after photorefractive keratectomy (PRK) to decrease postoperative pain caused by the movement of lids over the corneal epithelial defect and to facilitate epithelial healing. We assessed the microbiological profile of the conjunctiva of patients undergoing PRK for myopia, before and after the concurrent use of a therapeutic soft contact lens, and compared the effect on ocular bacterial colonization of prophylactic administration of topical tobramycin 0.3% versus ofloxacin 0.3%. Forty-three consecutive eyes from 37 patients underwent PRK for myopia or myopic astigmatism. Eyes were assigned randomly to prophylactic antibacterial treatment with either topical ofloxacin 0.3% or tobramycin 0.3%, applied prior to surgery and three times daily after surgery until therapeutic soft contact lens removal. Material from the conjunctival sac was obtained for bacteriologic cultures prior to surgery. Clinical evaluation of all eyes was conducted prospectively. Three days after PRK, the therapeutic soft contact lenses were removed and cultured. Cultures from the conjunctival sac were then repeated. No statistically significant differences were observed in culture positivity between the two groups of eyes, in spite of some positive preoperative and postoperative cultures. Only one out of 43 eyes (assigned to the ofloxacin group) developed a peripheral corneal infiltrate. The corneal infiltrate healed completely without sequelae using antibiotic and corticosteroid therapy. The use of therapeutic soft contact lenses after PRK with either topical tobramycin 0.3% or ofloxacin 0.3% were well tolerated. However, perioperative positive conjunctival cultures were relatively frequent and prophylactic antibiotics should be used in the setting of an epithelial defect and therapeutic soft contact lens following PRK.


Interpretation of respiratory functional explorations of deficiency and incapacity in adult.
Routine respiratory functional explorations (RFEs) performed as a part of the exploration of the respiratory deficiency include spirometry, plethysmography, bronchodilation test, measurement of the carbon monoxide diffusion capacity (DLCO), non-specific bronchial challenge (NSBC) and the measurement of the exhaled fraction of nitric oxide (FeNO). Functional exploration of disability is most often achieved by the 6-minute walk test (6MWT). All these RFEs were the subject of European and/or American consensus conferences in 2005 for the spirometry and plethysmography, in 2014 for the 6MWT, in 2015 for the FeNO, in 2017 for the NSBC and for the DLCO. The interpretation of the RFEs must therefore rationally be carried out according to the most recent international recommendations and requires reference to the lower and sometimes upper limits of normal (for spirometry, plethysmography, DLCO and 6-min walk distance), z-scores (for spirometry) or to fixed threshold values (for the bronchodilation test, FeNO and delivered dose of methacholine causing a 20% fall in forced expiratory volume in one second). Therefore, the main objective of this update was to expose the definitions and classifications of the various 'abnormalities' noted during the RFEs requested in the context of the exploration of the deficiency and incapacity of adult patients. This update will assist clinicians, especially pulmonologists, in the interpretation of the most commonly requested RFEs in practice.


Measurement of Clozapine, Norclozapine, and Amisulpride in Plasma and in Oral Fluid Obtained Using 2 Different Sampling Systems.
There is a poor correlation between total concentrations of proton-accepting compounds (most basic drugs) in unstimulated oral fluid and in plasma. The aim of this study was to compare clozapine, norclozapine, and amisulpride concentrations in plasma and in oral fluid collected using commercially available collection devices [Thermo Fisher Scientific Oral-Eze and Greiner Bio-One (GBO)]. Oral-Eze and GBO samples and plasma were collected in that order from patients prescribed clozapine. Analyte concentrations were measured by liquid chromatography-tandem mass spectrometry. There were 112 participants [96 men, aged (median, range) 47 (21-65) years and 16 women, aged 44 (21-65) years]: 74 participants provided 2 sets of samples and 7 provided 3 sets (overall 2 GBO samples not collected). Twenty-three patients were co-prescribed amisulpride, of whom 17 provided 2 sets of samples and 1 provided 3 sets. The median (range) oral fluid within the GBO samples was 52 (13%-86%). Nonadherence to clozapine was identified in all 3 samples in one instance. After correction for oral fluid content, analyte concentrations in the GBO and Oral-Eze samples were poorly correlated with plasma clozapine and norclozapine (R = 0.57-0.63) and plasma amisulpride (R = 0.65-0.72). Analyte concentrations in the 2 sets of oral fluid samples were likewise poorly correlated (R = 0.68-0.84). Mean (SD) plasma clozapine and norclozapine were 0.60 (0.46) and 0.25 (0.21) mg/L, respectively. Mean clozapine and norclozapine concentrations in the 2 sets of oral fluid samples were similar to those in plasma (0.9-1.8 times higher), that is, approximately 2- to 3-fold higher than those in unstimulated oral fluid. The mean (±SD) amisulpride concentrations (microgram per liter) in plasma (446 ± 297) and in the Oral-Eze samples (501 ± 461) were comparable and much higher than those in the GBO samples (233 ± 318). Oral fluid collected using either the GBO system or the Oral-Eze system cannot be used for quantitative clozapine and/or amisulpride therapeutic drug monitoring.


LR3M: Robust Low-Light Enhancement via Low-Rank Regularized Retinex Model.
Noise causes unpleasant visual effects in low-light image/video enhancement. In this paper, we aim to make the enhancement model and method aware of noise in the whole process. To deal with heavy noise which is not handled in previous methods, we introduce a robust low-light enhancement approach, aiming at well enhancing low-light images/videos and suppressing intensive noise jointly. Our method is based on the proposed Low-Rank Regularized Retinex Model (LR3M), which is the first to inject low-rank prior into a Retinex decomposition process to suppress noise in the reflectance map. Our method estimates a piece-wise smoothed illumination and a noise-suppressed reflectance sequentially, avoiding remaining noise in the illumination and reflectance maps which are usually presented in alternative decomposition methods. After getting the estimated illumination and reflectance, we adjust the illumination layer and generate our enhancement result. Furthermore, we apply our LR3M to video low-light enhancement. We consider inter-frame coherence of illumination maps and find similar patches through reflectance maps of successive frames to form the low-rank prior to make use of temporal correspondence. Our method performs well for a wide variety of images and videos, and achieves better quality both in enhancing and denoising, compared with the state-of-the-art methods.


A facile synthesis of a theranostic nanoparticle by oxidation of dopamine-DTPA-Gd conjugates.
Photothermal therapies (PPTs) with various light-absorbing materials have shown very promising therapeutic effects against cancers. However, their application was severely limited by the lack of accurate localization of tumors and real-time monitoring of the therapeutic process. Theranostic nanoparticles with both imaging and therapeutic functions are highly desired to develop imaging-mediated PPTs. Herein, we develop a facile one-pot method to synthesize a nanoparticle with functions of an MRI contrast agent and a PTT agent through oxidization of dopamine-DTPA-Gd conjugates and PEG-dopamine conjugates. The oxidized dopamine nanoparticles (ODNP) had a high R1 up to 9.6 mM-1 s-1, 2.2 times higher than that of Omniscan, and showed significantly higher MRI contrast enhancement than Omniscan in tumor. Meanwhile, the ODNP showed strong NIR light absorption and significant antitumor efficacy both in vitro and in vivo as a PPT agent. The ODNP with excellent MRI contrasting capability and PTT efficacy plus its facile synthesis and good biocompatibility are a very promising theranostic agent for MRI-mediated PTT.


Grazing management contributions to net global warming potential: a long-term evaluation in the Northern Great Plains.
The role of grassland ecosystems as net sinks or sources of greenhouse gases (GHGs) is limited by a paucity of information regarding management impacts on the flux of nitrous oxide (N(2)O) and methane (CH(4)). Furthermore, no long-term evaluation of net global warming potential (GWP) for grassland ecosystems in the northern Great Plains (NGP) of North America has been reported. Given this need, we sought to determine net GWP for three grazing management systems located within the NGP. Grazing management systems included two native vegetation pastures (moderately grazed pasture [MGP], heavily grazed pasture [HGP]) and a heavily grazed crested wheatgrass [Agropyron desertorum (Fisch. ex. Link) Schult.] pasture (CWP) near Mandan, ND. Factors evaluated for their contribution to GWP included (i) CO(2) emissions associated with N fertilizer production and application, (ii) literature-derived estimates of CH(4) production for enteric fermentation, (iii) change in soil organic carbon (SOC) over 44 yr using archived soil samples, and (iv) soil-atmosphere N(2)O and CH(4) fluxes over 3 yr using static chamber methodology. Analysis of SOC indicated all pastures to be significant sinks for SOC, with sequestration rates ranging from 0.39 to 0.46 Mg C ha(-1) yr(-1). All pastures were minor sinks for CH(4) (<2.0 kg CH(4)-C ha(-1) yr(-1)). Greater N inputs within CWP contributed to annual N(2)O emission nearly threefold greater than HGP and MGP. Due to differences in stocking rate, CH(4) production from enteric fermentation was nearly threefold less in MGP than CWP and HGP. When factors contributing to net GWP were summed, HGP and MGP were found to serve as net CO(2equiv.) sinks, while CWP was a net CO(2equiv.) source. Values for GWP and GHG intensity, however, indicated net reductions in GHG emissions can be most effectively achieved through moderate stocking rates on native vegetation in the NGP.


Papillary renal cell-like carcinoma in a retroperitoneal teratoma.
We report a case of somatic type malignancy with papillary renal cell carcinoma differentiation arising in a retroperitoneal mixed germ cell tumor. The patient was a 36-year-old man with a synchronous mediastinal teratoma. The somatic type malignancy in the retroperitoneal tumor was composed of papillary structures covered by atypical epithelial cells with eosinophilic cytoplasm, prominent nucleoli and pseudostratified nuclei. Papillary cores contained numerous aggregates of foamy macrophages, typical of type I papillary renal cell carcinomas. The immunohistochemical profile was consistent with papillary renal cell carcinoma, including positive reactions for cytokeratin 7 and alpha-methyl acyl CoA racemase. There was no somatic type malignancy component in the synchronous mediastinal teratoma. Both the retroperitoneal and the mediastinal tumor showed gains of 12p and chromosome 17 material. There was no c-MET mutation in the somatic type malignancy. To our knowledge, this is the first report of a somatic type malignancy with features of papillary renal cell carcinoma arising in a germ cell tumor. It is important not to confuse such a retroperitoneal tumor with a conventional papillary renal cell carcinoma, because presence of other malignant histologies within the germ cell tumor may warrant different treatment. In such cases, the presence of isochromosome 12p can be helpful to the diagnosis.


Glycemia Regulation: From Feedback Loops to Organizational Closure.
Endocrinologists apply the idea of feedback loops to explain how hormones regulate certain bodily functions such as glucose metabolism. In particular, feedback loops focus on the maintenance of the plasma concentrations of glucose within a narrow range. Here, we put forward a different, organicist perspective on the endocrine regulation of glycaemia, by relying on the pivotal concept of closure of constraints. From this perspective, biological systems are understood as organized ones, which means that they are constituted of a set of mutually dependent functional structures acting as constraints, whose maintenance depends on their reciprocal interactions. Closure refers specifically to the mutual dependence among functional constraints in an organism. We show that, when compared to feedback loops, organizational closure can generate much richer descriptions of the processes and constraints at play in the metabolism and regulation of glycaemia, by making explicit the different hierarchical orders involved. We expect that the proposed theoretical framework will open the way to the construction of original mathematical models, which would provide a better understanding of endocrine regulation from an organicist perspective.


Dysmorphogenesis of lymph nodes in Foxc2 haploinsufficient mice.
Dysmorphogenesis of lymph nodes displayed in a fork head transcription factor Foxc2 haploinsufficient mice--a model for lymphedema-distichiasis syndrome--was studied by immunohistochemistry and electron microscopy. The Foxc2 heterozygous mice manifested lymph node hyperplasia composed of conspicuous proliferation of endothelial cells forming the lymphatic sinus and α-smooth muscle actin (SMA)-immunopositive fibroblast-like cells in the lymphatic pulp, particularly around the sinus. The hyperplastic sinus endothelial cells and the SMA-positive cells demonstrated distinct immunolocalization of platelet-derived growth factor (PDGF)-B, a crucial chemoattractant for vascular mural cell recruitment, and its receptor, PDGFR-β, respectively. The observations suggest that the sinus endothelial cells elicit abnormal recruitment of the fibroblast-like cells as a type of vascular mural cells via PDGF-B/PDGFR-β signaling in lymph nodes of the Foxc2 heterozygotes. Furthermore, in Foxc2 heterozygous lymph nodes, recruited SMA-positive cells displayed an intense immunoreaction for vascular endothelial growth factor (VEGF)-C, a highly specific lymphangiogenic factor, and its receptor, VEGFR-3, was preferentially distributed in the lymphatic sinus endothelial cells. These findings suggest that an interactive cycle between lymphatic sinus endothelial cells and the fibroblast-like cells, which involves PDGF-B/PDGFR-β and VEGF-C/VEGFR-3 signaling, is essential for aberrant hyperplasia of the lymphatic sinus and the fibroblast-like cells in Foxc2 haploinsufficiency.


Usefulness of BP180 NC16a enzyme-linked immunosorbent assay in the serodiagnosis of pemphigoid gestationis and in differentiating between pemphigoid gestationis and pruritic urticarial papules and plaques of pregnancy.
Pemphigoid gestationis (PG) is a rare pregnancy-associated subepidermal immunobullous disease that targets hemidesmosomal proteins, particularly BP180. Clinically, PG can resemble the eruption known as polymorphic urticarial papules and plaques of pregnancy (PUPPP), and accurate differentiation between these 2 pruritic pregnancy dermatoses has important implications for fetal and maternal prognoses. Results of epitope mapping studies show that IgG autoantibodies in up to 90% of PG serum samples target the well-defined membrane-proximal NC16a domain of BP180. To examine the usefulness of a commercially available NC16a domain enzyme-linked immunosorbent assay in the serodiagnosis of PG and in the differentiation of PG from PUPPP. A total of 412 women consisting of pretreatment patients with PG (n = 82), patients with PUPPP (n = 164), and age- and sex-matched controls (n = 166). All serum samples were assayed in duplicate. Receiver operating characteristic analyses were performed to determine a cutoff value for the diagnosis of PG and for differentiation from PUPPP and controls. A cutoff value of 10 enzyme-linked immunosorbent assay units was associated with specificity and sensitivity of 96%. The NC16a enzyme-linked immunosorbent assay is highly sensitive and highly specific in differentiating PG from PUPPP, and it is potentially a valuable tool in the serodiagnosis of PG.


Fecal source tracking, the indicator paradigm, and managing water quality.
Fecal source tracking is used because standard methods of measuring fecal contamination in water by enumerating fecal indicator bacteria (FIB) do not identify the sources of the contamination. This paper presents a critical review of source tracking with emphasis on the extent to which methods have been tested (especially in comparison with other methods and/or with blind samples), when methods are applicable, their shortcomings, and their usefulness in predicting public health risk or pathogen occurrence. In addition, the paper discusses the broader question of whether fecal source tracking and fecal indicator monitoring is the best approach to regulate water quality and protect human health. Many fecal source-tracking methods have only been tested against sewage or fecal samples or isolates in laboratory studies (proof of concept testing) and/or applied in field studies where the "real" answer is not known, so their comparative performance and accuracy cannot be assessed. For source tracking to be quantitative, stability of ratios between host-specific markers in the environment must be established. In addition, research is needed on the correlation between host-specific markers and pathogens, and survival of markers after waste treatments. As a result of the exclusive emphasis on FIB in legislation, monitoring has concentrated on FIB and lost sight of pathogens. A more rational approach to regulating water quality would start with available epidemiological data to identify pathogens of concern in a particular water body, and then use targeted pathogen monitoring coupled with targeted fecal source tracking to control them. Baseline monitoring of indicators would become just one tool among many.


[Inactivating effect of N-methyl-N'-nitro-N-nitrosoguanidine on yeasts with different ploidies].
Haploid and diploid strains of Saccharomyces cerevisiae, Sacch. ellipsoideus and Pichia pinus were studied. Differences in N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) sensitivity were detected both between haploids and diploids of the same species and between the corresponding strains of different species. Survival curves after MNNG treatment of all strains irrespectively of ploidy were exponential with "a tail". All the strains also exhibited the delayed appearance of clones from MNNG-treated cells. Three different forms of cell inactivation after MNNG treatment were detected similar to those observed after irradiation.


Modulation of apoptosis by cigarette smoke and cancer chemopreventive agents in the respiratory tract of rats.
Preclinical studies may elucidate the meaning of biomarkers applicable to epidemiologic studies and to clinical trials for cancer prevention. No study has explored so far the effect of cigarette smoke on apoptosis in vivo. We evaluated modulation of apoptosis in cells of the respiratory tract of smoke-exposed Sprague-Dawley rats both by morphological analysis and TUNEL method. In a first study, exposure of rats to mainstream cigarette smoke for either 18 or 100 consecutive days produced a significant and time-dependent increase in the proportion of apoptotic cells in the bronchial and bronchiolar epithelium. Oral N:-acetylcysteine did not affect the background frequency of apoptosis but significantly and sharply decreased smoke-induced apoptosis. In a second study, exposure of rats to a mixture of sidestream and mainstream smoke for 28 consecutive days resulted in a >10-fold increase in the frequency of pulmonary alveolar macrophages undergoing apoptosis. Dietary administration of either 5,6-benzoflavone, 1,2-dithiole-3-thione or oltipraz did not affect the frequency of smoke-induced apoptosis, whereas phenethyl isothiocyanate produced a further significant enhancement. Again, N-acetylcysteine and its combination with oltipraz significantly decreased smoke-induced apoptosis. In both studies exposure to smoke resulted in a sharp increase of cells positive for proliferating cell nuclear antigen (PCNA), which was unaffected by the examined chemopreventive agents. These findings highlight the concept that modulation of apoptosis has diversified meanings. Different meanings (as explained in the following lines). First, the apoptotic process is triggered as a defense system against genotoxic agents, such as the components of cigarette smoke. The further induction produced by phenethyl isothiocyanate, favoring removal of damaged cells, represents an example of a detoxification mechanism. Inhibition of smoke-induced apoptosis by N:-acetylcysteine should be interpreted as an epiphenomenon of antigenotoxic mechanisms, as shown in parallel studies evaluating modulation of DNA alterations in the respiratory tract of the same animals. Thus, it is important to discriminate between whether the opposite modulation of apoptosis is per se a protective mechanism or the beneficial outcome of other mechanisms inhibiting genotoxicity.


Dracunculiasis in Igbon, Oyo State, Nigeria.
Studies conducted between 1978 and 1979 to determine the prevalence and site of guinea-worm transmission in Igbon, Nigeria, showed that in less than 5 years of the introduction of dracunculiasis into this community, the infection rate had reached over 75% among the economically active group. Multiple infections were rampant and the frequency of occurrence of severe incapacitation was correspondingly high as reflected by prolonged and excessive absenteeism from schools, inability to engage in farming, trading and domestic work. The peak of transmission occurred in November which coincided with the retreat of rain and the abundance of Thermocyclops nigerianus in the two adjacent ponds where active transmission occurred. While ignorance was primarily responsible for annual transmission, secondary infection resulting from unsanitary methods of local treatment, seemed to be the major cause of prolonged and severe incapacitation.


Thyroglobulin biosynthesis in "cold" and "hot" nodules in the human thyroid gland.
Studies on "cold" and "hot" nodules from human thyroid gland have been carried out to investigate thyroglobulin biosynthesis and to correlate carbohydrates incorporation and thyroid hormone formation in thyroglobulin. The aim of the investigation was to ascertain the role of carbohydrates of the protein molecule in its migration to the iodinating site of the cell. Thyroid slices from two cases with "cold" and two with "hot" nodules were incubated for 30 to 120 min with [3H]leucine, [3H]carbohydrates (ManNAc, Gal, Man, GlcNAc, GalNAc) and 125I. Soluble and particulate iodoproteins, solubilized by digitonin, were identified by density gradient centrifugation and immunoprecipitation; following hydrolysis carbohydrates and iodothyronines were identified by paper chromatography and chemically determined. While the rate of leucine incorporation into TG increased with time in both "cold" and "hot" nodules, the "cold" nodule, with respect to the "hot" tissue, showed: much lower efficiency incorporating carbohydrates and iodine into TG; lower thyroid/medium ratio of radioiodine; less total soluble protein and soluble TG; more particulate protein and solubilized TG; 1/3 of the carbohydrate and iodine chemical content; only minute amounts of labeled iodothyronines in TG compared to normal levels in "hot" tissue. The formation of thyroid hormone, in enzymatically iodinated TG, and the iodinating activity of the 105,000 X g pellet, incubated with low iodine TG, were similar in both "cold" and "hot" tissues. These results demonstrate that the "cold" nodule is able to synthesize thyroglobulin but the protein is defective in its carbohydrate content. The deficient iodine transport into "cold" tissue and impaired hormonal synthesis are confirmed. The incomplete incorporation of carbohydrates into thyroglobulin in the "cold" nodule could be important for the maturation and migration of the molecule to the iodinating site of the cell


Dipole preserving and polarization consistent charges.
A method for estimating dipole preserving and polarization consistent (DPPC) charges is described, which reproduces exactly the molecular dipole moment as well as the local, atomic hybridization dipoles determined from the corresponding wave function and can yield accurate molecular polarization. The method is based on a model described by Thole and van Duijnen and a new feature is introduced to treat molecular polarization. Thus, the DPPC method offers a convenient procedure to describe molecular polarization in applications using semiempirical models and ab initio molecular orbital theory with relatively small basis functions such as 6-31+G(d,p) or without inclusion of electron correlation; these methods tend to underestimate molecular polarizability. The trends of the DPPC partial atomic charges are found to be in good accord with those of the CM2 model, a class IV charge analysis method that has been used in a variety of applications. The DPPC method is illustrated to mimic the correct molecular polarizability in a water dimer test case and in water-halide ion complexes using the explicit polarization (X-Pol) potential with the Austin model 1 Hamiltonian.


HIV-(1/2) indeterminate western blot results: follow-up of asymptomatic blood donors in belo horizonte, minas gerais, brazil
The clinical and public health importance of indeterminate results in HIV-(1/2) testing is still difficult to evaluate in volunteer blood donors. At Fundacao Hemominas, HIV-(1/2) ELISA is used as the screening test and, if reactive, is followed by Western blot (WB). We have evaluated 84 blood donors who had repeatedly reactive ELISA tests for HIV-(1/2), but indeterminate WB results. Sixteen of the 84 donors (19.0%) had history of sexually transmitted diseases; 18/84 (21.4%) informed receiving or paying for sex; 3/84 (3.6%) had homosexual contact; 2/26 women (7.6%) had past history of multiple illegal abortions and 3/84 (3.6%) had been previously transfused. Four out of 62 donors (6.5%) had positive anti-nuclear factor (Hep2), with titles up to 1:640. Parasitological examination of the stool revealed eggs of S. mansoni in 4/62 (6.4%) donors and other parasites in 8/62 (12.9%). Five (5.9%) of the subjects presented overt seroconversion for HIV-(1/2), 43/84 (51.2%) had negative results on the last visit, while 36/84 (42.9%) remained WB indeterminate. Although some conditions could be found associated with the HIV-(1/2) indeterminate WB results and many donors had past of risky behavior, the significance of the majority of the results remains to be determined.


Gel-in-water nanodispersion for potential application in intravenous delivery of anticancer drugs.
Organogels are semi-solid systems that can gel organic liquids at low concentrations. The use of organogels in drug delivery has grown rapidly in the last decade owing to their fibrous microstructure and suitability for different routes of administration. The current study is characterized by nanogel dispersion (NGD) development based on emulsion technology. The efficiency of this organogel based NGD as a carrier for anticancer drugs was assessed both in vitro and in vivo. 12-Hydroxystearic acid formed an organogel with lipiodol and encapsulated the anticancer drug paclitaxel. The gel-in-water (G/W) nanodispersion was prepared via ultrasonication and stabilized by a nonionic surfactant. The results showed that the organogel enabled sustained drug release from G/W nanodispersion over time, along with enhanced cellular uptake. The prepared G/W nanodispersion was found to be biocompatible with mouse hepatocytes and fibroblast cells in vitro, whereas paclitaxel-loaded G/W nanodispersion showed cytotoxicity (p <0.05) against lung cancer (A549) cell lines. Similarly, intravenous administration of paclitaxel-loaded G/W nanodispersion exerts an anticancer effect against lung cancer in vivo, with a significant decrease in tumor volume (p <0.05). Therefore, the proposed G/W nanodispersion could be a promising carrier for chemotherapy agents with sustained drug release and better therapeutic outcomes against cancer.


Factors associated with vaccine intention in adults living in England who either did not want or had not yet decided to be vaccinated against COVID-19.
Early studies showed that 28-36% of UK adults were unsure or unwilling to be vaccinated against COVID-19. We wanted to identify which socio-demographic, socio-economic, personal health and psychological factors were associated with COVID-19 vaccine intentions (CVI) in adults living in England who did not want, yet to consider, or not sure whether to vaccinate. In October/November 2020, prior to vaccine availability, we surveyed adults stratified by gender, region, and deprivation, with additional purposive sampling of those aged 50 and over and those from an ethnic minority. Two hundred and ten did not want; 407 had yet to consider; and 1,043 were not sure whether to be vaccinated. Factors positively associated with CVI were: favorable vaccine views, trust in institutions associated with vaccine approval, vaccine subjective norms, anticipated regret of not having a vaccine, perceived vaccine benefits, perceived safety knowledge sufficiency, and a history of having an influenza vaccine. Factors negatively associated were: anti-lockdown views, and being a health or social care worker. Whilst showing significant relationships with CVI when analyzed in isolation, neighborhood deprivation and ethnicity did show an independent relationship to intention when all study measures were controlled for. Our findings suggest vaccine promotion focusing on the anticipated regret of not having a vaccine, the benefits of a mass COVID-19 immunization program, and the safety of a vaccine whilst ensuring or engendering trust in those bodies that brand a campaign may be most supportive of COVID-19 vaccine uptake.


Computer assisted instruction on "learning nutrition flags for deaf 5th grade and 6th grad students": effectiveness of instruction.
Deaf students are of a number of under privilege group for whom there are limited resources for their use, related to health including nutrition. The purpose of this research was to create computer-assisted instruction for "nutrition flags" for 5 and 6th grade students. The content of nutrition included the concept of a healthy balance diets and portion sizes of each food group. The content and pictures for computer-assisted instruction came from existing curriculum, and focused on nutritional content. The contents in this instruction were divided into three units according to students' learning capacity. The story boards were developed by staff including nutritionists, Thai sign language interpreters, and deaf students. Then, the contents and nutrition vocabulary were translated into Thai sign language. After recording the sign language on video, this material was merged with the contents and converted into a computer program. The computer assisted instruction was tested with students from Nakon Pathom School for the Deaf The first trial was conducted with three students, the second with five students, and the third with 15 students during the academic year 2009. The computer- assisted instruction was revised until it met the standard criteria of 80/80. Effectiveness testing was carried out with 36 students for five consecutive days. On the first day, the pre-test was completed, and on days 2-4, the students performed self-study and completed the exercises for units 1-3, with 50 minutes spent on each unit. The post-test was completed on the last day. The study was conducted during the 2010 academic year Data analysis was performed using the t-test. The results showed an effectiveness of 81.85/82.22, which was higher than the standard criteria of 80/80. The post-test average score was higher than the pre-test average score with a statistical significance level at p < 0.0001. Suggestions for instruction for the deaf are that the length of the instruction in each unit should be no longer than 30 minutes and the sign language window should be large because this mode of communication relies on sight.


Rorschach score validation as a model for 21st-century personality assessment.
Recent conceptual and methodological innovations have led to new strategies for documenting the construct validity of test scores, including performance-based test scores. These strategies have the potential to generate more definitive evidence regarding the validity of scores derived from the Rorschach Inkblot Method (RIM) and help resolve some long-standing controversies regarding the clinical utility of the Rorschach. After discussing the unique challenges in studying the Rorschach and why research in this area is important given current trends in scientific and applied psychology, I offer 3 overarching principles to maximize the construct validity of RIM scores, arguing that (a) the method that provides RIM validation measures plays a key role in generating outcome predictions; (b) RIM variables should be linked with findings from neighboring subfields; and (c) rigorous RIM score validation includes both process-focused and outcome-focused assessments. I describe a 4-step strategy for optimal RIM score derivation (formulating hypotheses, delineating process links, generating outcome predictions, and establishing limiting conditions); and a 4-component template for RIM score validation (establishing basic psychometrics, documenting outcome-focused validity, assessing process-focused validity, and integrating outcome- and process-focused validity data). The proposed framework not only has the potential to enhance the validity and utility of the RIM, but might ultimately enable the RIM to become a model of test score validation for 21st-century personality assessment.


Heterotaxy - Res ipsos loquitur.
The essence of so-called heterotaxy is the potential disharmony between the arrangement of the bronchuses, abdominal organs, and the atrial appendages. Accurate description of the heart, however, can only be provided by specific description of these features, all of which are readily discernible in the clinical setting. We argue that, when accurate description of the atrial and visceral arrangement is provided, along with appropriate description of the intracardiac findings, no further accuracy is gained by suggesting that an individual heart is "heterotaxic".


Safety and efficacy of the Derivo Embolization Device for the treatment of unruptured intracranial aneurysms: a multicentric study.
The Derivo Embolization Device (DED) is a novel flow diverter stent that provides increased x-ray visibility, an improved delivery system, and potentially reduced thrombogenicity. The objective of this study was to evaluate the early safety and efficacy of the second-generation DED. We retrospectively analyzed all patients with unruptured intracranial aneurysms (UIAs) treated with the DED between November 2015 and December 2017 in three German tertiary care centers. Procedural details, complications, and morbidity within 30 days after treatment, as well as the aneurysm occlusion rates after 6 months (O'Kelly-Marotta scale, OKM), were evaluated. Implantation of the DED was attempted in 42 patients with 42 aneurysms. All procedures were technically successful. Multiple DEDs were used in three aneurysms (7.2%) and adjunctive coiling in 11 (26.2%). Procedure-related complications occurred in four cases (9.5%) including three thromboembolic events and one aneurysm perforation. The morbidity rate was 2.4% and there was no mortality. One patient suffered an ischemic stroke with persistent aphasia at 30-day follow-up due to a thromboembolic infarct (modified Rankin Scale score 1). Among 33 patients (78.6%) available for angiographic follow-up, complete (OKM D) and favorable (OKM C+D) aneurysm occlusion was obtained in 72.7% (24/33) and 87.9% (29/33), respectively. Endovascular treatment of UIAs with the DED is associated with high procedural safety and adequate occlusion rates. Examinations at 1- and 2-year follow-up will provide data on the long-term safety and angiographic outcomes of this device.


Involvement of the NGFI-A gene in the differentiation of neuroblastoma cells.
The transcription factor NGFI-A is an early response gene that has been implicated in the regulation of cell growth and differentiation and, more recently, in apoptosis. This gene is expressed in many tissues, and is very abundant in the brain. However, little is known about its functional role in the differentiation of this tissue. In the present work we investigated the role of NGFI-A in serum withdrawal-induced differentiation in N2A neuroblastoma cells. To do so, we studied the effect of NGFI-A antisense oligonucleotides and NGFI-A overexpression on this process. We show that neuroblastoma cells treated with an NGFI-A antisense oligonucleotide do not undergo normal morphological differentiation after serum withdrawal, whereas N2A cells overexpressing this gene extend long neurites, even in the presence of serum. We also show that NGFI-A overexpression is accompanied by an increase in the amount of phosphorylated microtubule-associated protein MAP1B, which has been associated with neurite outgrowth. Our results suggest that the NGFI-A gene plays an important role in neurite extension.


Immunological reaction in TNF-α-mediated osteoclast formation and bone resorption in vitro and in vivo.
Tumor necrosis factor- α (TNF- α ) is a cytokine produced by monocytes, macrophages, and T cells and is induced by pathogens, endotoxins, or related substances. TNF- α may play a key role in bone metabolism and is important in inflammatory bone diseases such as rheumatoid arthritis. Cells directly involved in osteoclastogenesis include macrophages, which are osteoclast precursor cells, osteoblasts, or stromal cells. These cells express receptor activator of NF- κ B ligand (RANKL) to induce osteoclastogenesis, and T cells, which secrete RANKL, promote osteoclastogenesis during inflammation. Elucidating the detailed effects of TNF- α on bone metabolism may enable the identification of therapeutic targets that can efficiently suppress bone destruction in inflammatory bone diseases. TNF- α is considered to act by directly increasing RANK expression in macrophages and by increasing RANKL in stromal cells. Inflammatory cytokines such as interleukin- (IL-) 12, IL-18, and interferon- γ (IFN- γ ) strongly inhibit osteoclast formation. IL-12, IL-18, and IFN- γ induce apoptosis in bone marrow cells treated with TNF- α in vitro, and osteoclastogenesis is inhibited by the interactions of TNF- α -induced Fas and Fas ligand induced by IL-12, IL-18, and IFN- γ . This review describes and discusses the role of cells concerned with osteoclast formation and immunological reactions in TNF- α -mediated osteoclastogenesis in vitro and in vivo.


Modelling the central nervous system: tissue engineering of the cellular microenvironment.
With the increasing prevalence of neurodegenerative diseases, improved models of the central nervous system (CNS) will improve our understanding of neurophysiology and pathogenesis, whilst enabling exploration of novel therapeutics. Studies of brain physiology have largely been carried out using in vivo models, ex vivo brain slices or primary cell culture from rodents. Whilst these models have provided great insight into complex interactions between brain cell types, key differences remain between human and rodent brains, such as degree of cortical complexity. Unfortunately, comparative models of human brain tissue are lacking. The development of induced Pluripotent Stem Cells (iPSCs) has accelerated advancement within the field of in vitro tissue modelling. However, despite generating accurate cellular representations of cortical development and disease, two-dimensional (2D) iPSC-derived cultures lack an entire dimension of environmental information on structure, migration, polarity, neuronal circuitry and spatiotemporal organisation of cells. As such, researchers look to tissue engineering in order to develop advanced biomaterials and culture systems capable of providing necessary cues for guiding cell fates, to construct in vitro model systems with increased biological relevance. This review highlights experimental methods for engineering of in vitro culture systems to recapitulate the complexity of the CNS with consideration given to previously unexploited biophysical cues within the cellular microenvironment.


Medial Prefrontal Cortex Represents the Object-Based Cognitive Map When Remembering an Egocentric Target Location.
A cognitive map, representing an environment around oneself, is necessary for spatial navigation. However, compared with its constituent elements such as individual landmarks, neural substrates of coherent spatial information, which consists in a relationship among the individual elements, remain largely unknown. The present study investigated how the brain codes map-like representations in a virtual environment specified by the relative positions of three objects. Representational similarity analysis revealed an object-based spatial representation in the hippocampus (HPC) when participants located themselves within the environment, while the medial prefrontal cortex (mPFC) represented it when they recollected a target object's location relative to their self-body. During recollection, task-dependent functional connectivity increased between the two areas implying exchange of self-location and target location signals between the HPC and mPFC. Together, the object-based cognitive map, whose coherent spatial information could be formed by objects, may be recruited in the HPC and mPFC for complementary functions during navigation, which may generalize to other aspects of cognition, such as navigating social interactions.