A one year audit of the Colorectal Unit at Wits Donald Gordon Medical Centre: 2016-2017.
Colorectal surgery has developed into an established surgical subspecialty in South Africa, however there is a paucity of data regarding the epidemiology and surgical outcomes of patients with colorectal disease in this country. The objective is to present the findings of a one-year audit of the Wits Donald Gordon Medical Centre (WDGMC) Colorectal Unit with specific reference to indications, surgical procedures and patient outcomes. Patient files from December 2016 to November 2017 were included in a retrospective analysis. The Mann-Whitney U test was used to analyse continuous variables and the Chi-squared test was used to compare categorical variables. During the audit period, 1264 patients were admitted to the Colorectal Unit and a further 564 outpatient endoscopic procedures were performed. There were 306 emergency admissions. 139 elective colorectal resections took place, with a 16% major complication rate, a 12% anastomotic leak rate and no deaths. Rectal resections constituted 66% of the operations and 34% were colonic resections. The median length of stay for all patients undergoing resection was 9 days and there was no statistically significant difference in length of stay between open and laparoscopic cases. The WDGMC Colorectal Unit manages a high volume of patients presenting with the full spectrum of colorectal disease.


Distribution of Sorangium cellulosum.
The myxobacterium Sorangium cellulosum exists and plays an important role in soil ecology by its ability to degrade cellulosic materials; it was found to be widely distributed in North American soils.


Comparison of the baseline disease activity of early oligo- and polyarthritis in sequential years.
Many early arthritis clinics (EACs) have been started in the last decade in order to detect and treat rheumatoid arthritis early. The present study evaluates whether the disease activity at admission of patients with early oligo- and polyarthritis changed during the period 1993--1998 in two EACs in the Netherlands. Patients were selected who were diagnosed after one year as having rheumatoid arthritis (RA) or oligo- or polyarthritis (UPA), had a symptom duration of less than 2 years, and were referred from two Dutch EACs between 1993 and 1998. The data from the two clinics were combined and stratified by referral year. Differences in baseline disease characteristics as well as changes in radiological and functional scores after two years of follow-up between referral years were analysed by ANOVA using Bonferroni corrected p levels. A total of 405 patients (66% females; median age 57 yrs (18-93): 80% diagnosed as RA, the remainder as UPA) were included in the study. The year-groups did not differ significantly in demographic characteristics or in the duration of complaints (median 6 months). The number of patients with a diagnosis of RA declined over the years, as did the mean baseline erythrocyte sedimentation rate (ESR), in RA and UPA patients. The functional status (Health Assessment Questionnaire: HAQ) was enhanced in 1998 compared with the previous years (p < 0.001). Radiographic progression (Sharp/van der Heijde score) after the 2-year follow-up decreased (p < 0.001) in the later referral years compared to the referral group of 1994. Disease modifying anti-rheumnatic drugs (DMARDs) were started in an earlier stage and the prescription rate of sulfasalazine and methotrexate increased over the years, whereas the number of patients not treated with DMARDs declined. The pattern of patient referral changed over 6 years towards fewer patients who fulfilled the RA diagnosis and a lower ESR (among UPA as well as RA patients), whereas the number of swollen joints and the duration of complaints remained the same. The radiological progression declined over time, probably due to less inflammation at the first visit and the increased use of DMARDs.


Phylogenetic Analyses Suggest that Factors Other Than the Capsid Protein Play a Role in the Epidemic Potential of GII.2 Norovirus.
Norovirus is the leading cause of acute gastroenteritis worldwide. For over two decades, a single genotype (GII.4) has been responsible for most norovirus-associated cases. However, during the winter of 2014 to 2015, the GII.4 strains were displaced by a rarely detected genotype (GII.17) in several countries of the Asian continent. Moreover, during the winter of 2016 to 2017, the GII.2 strain reemerged as predominant in different countries worldwide. This reemerging GII.2 strain is a recombinant virus that presents a GII.P16 polymerase genotype. In this study, we investigated the evolutionary dynamics of GII.2 to determine the mechanism of this sudden emergence in the human population. The phylogenetic analyses indicated strong linear evolution of the VP1-encoding sequence, albeit with minor changes in the amino acid sequence over time. Without major genetic differences among the strains, a clustering based on the polymerase genotype was observed in the tree. This association did not affect the substitution rate of the VP1. Phylogenetic analyses of the polymerase region showed that reemerging GII.P16-GII.2 strains diverged into a new cluster, with a small number of amino acid substitutions detected on the surface of the associated polymerase. Thus, besides recombination or antigenic shift, point mutations in nonstructural proteins could also lead to novel properties with epidemic potential in different norovirus genotypes. IMPORTANCE Noroviruses are a major cause of gastroenteritis worldwide. Currently, there is no vaccine or specific antiviral available to treat norovirus disease. Multiple norovirus strains infect humans, but a single genotype (GII.4) has been regarded as the most important cause of viral gastroenteritis outbreaks worldwide. Its persistence and predominance have been explained by the continuous replacement of variants that present new antigenic properties on their capsid protein, thus evading the herd immunity acquired to the previous variants. Over the last three seasons, minor genotypes have displaced the GII.4 viruses as the predominant strains. One of these genotypes, GII.2, reemerged as predominant during 2016 to 2017. Here we show that factors such as minor changes in the polymerase may have driven the reemergence of GII.2 during the last season. A better understanding of norovirus diversity is important for the development of effective treatments against noroviruses.


Timing and patterns of exposures during pregnancy and their implications for study methods.
Patterns of exposure variability across pregnancy were examined for medical, lifestyle, residential, and occupational exposures in a population-based sample of 357 livebirths from 10 rural California counties. A new measure of variability, the ratio of overall prevalence to time-window-specific prevalence, is introduced. The higher the overall : time window (OTW) ratio, the greater the potential for misclassification when using anytime-during-pregnancy prevalence for an agent that exerts its effect in a smaller time window. Exposures to cigarettes, marijuana, saunas/hot tubs, factors related to location of residence, and some workplace substances tended to be of longer duration. Intertrimester concordance was high (kappa's > 0.8) for smoking, residential proximity to crops, and use of video display terminals; moderately high (kappa's between 0.4 and 0.8) for many occupational exposures; and low (kappa's < 0.4) for illnesses, which tended to be of short duration. The lowest OTW ratios were for smoking and some residential exposures (1.1-1.3), while OTW ratios were much higher for paint applications, influenza, vaginal infections, and ultrasound (reaching, e.g., 4-6). Use of anytime-during-pregnancy exposure indices can bias measures of association between risk factors and adverse pregnancy outcomes, particularly if the OTW ratio is high. Misclassification bias occurs if there is a vulnerable time window during which the exposure exerts its effect. The misclassification can be differential when the average length of gestation of cases is shorter than that of controls. For exposures that vary, investigations of pregnancy outcome should collect as much detail as feasible regarding timing.


Pokeweed mitogen-, concanavalin A-, and phytohemagglutinin-induced development of cytotoxic effector lymphocytes. An evaluation of the mechanisms of T cell-mediated cytotoxicity.
Cultures of chicken lymphoid tissues were tested for their capacity to lyse (51)Cr-labeled chicken, burro (BRC), and human red blood cells (HRC) in the presence of phytomitogens. PHA-stimulated cultures lysed all three types of targets, while PWM and Con A showed a "target cell specificity" for HRC and BRC, respectively. In mixtures of target cells only the appropriate targets were lysed by lymphocytes activated by either Con A or PWM indicating that soluble lymphotoxins do not play a major role in these reactions. Preincubation experiments suggested that there may be a population of pre-existing aggressor cells which only require linking to the targets by the mitogens for activation of their cytotoxic potential. Strong cytotoxic reactions were found with spleen cells, peripheral blood leucocytes, and bone marrow cells. Thymocytes were less active but could be stimulated for significant cytotoxicity, while bursal cells were generally unreactive. Spleen cells from agammaglobulinemic chickens totally lacking serum immunoglobulins and B cells with surface-bound immunoglobulins were as active as cells from normal chickens. The activity of spleen cells, from which phagocytic cells were removed was also unimpaired. These results indicate that the development of cytotoxic effector lymphocytes in mitogen-treated leucocyte cultures is a property of T lymphocytes. Although bone marrow cells fail to proliferate in response to these phytomitogens, they do have strong cytotoxic reactivity suggesting that different subsets of thymic-derived lymphocytes are responsible for mitogen-induced transformation and mitogen-induced cytotoxicity.


Bioelectrical impedance plethysmographic analysis of body composition in critically injured and healthy subjects.
Determination of body composition during critical illness is complex because of various patient-related and technical factors. Bioelectrical impedance is a promising technique for the analysis of body composition; however, its clinical utility in critically injured patients is unknown. The purpose of this study was to compare bioelectrical impedance with metabolic activity in healthy and critically injured patients. If bioelectrical impedance accurately determines body composition during critical illness, the slope between body-composition variables and oxygen consumption would be the same in critically injured and healthy subjects. There is a strong linear relation between body composition and metabolic activity. In the present study, body composition (fat-free mass and body cell mass) was determined by using bioelectrical impedance and resting metabolic activity (metabolic rate and oxygen consumption) by using gas exchange analysis in a group of healthy and critically injured subjects. The relation between these variables was compared by using linear regression to a similar relation established by hydrostatic weighing in a large historical control group. The slope of the line relating fat-free mass to resting metabolic rate was the same in the healthy and critically ill groups (P = 0.62) and each was similar to the slope of the line for the control group. However, in 37% of the critically injured group, overhydration contributed to an increase in fat-free mass, disturbing the relation with resting metabolic rate. The slope of the line relating body cell mass to oxygen consumption in our healthy and critically ill groups was almost identical. These results support the use of bioelectrical impedance to determine body cell mass in healthy and critically ill subjects.


Adrenal venous sampling for primary aldosteronism and clinical outcomes after unilateral adrenalectomy: a single-center experience.
Adrenal venous sampling (AVS) remains controversial in the management of primary aldosteronism. Retrospective chart review was conducted at the Hospital of the University of Pennsylvania from July 2001 to September 2007. A total of 113 patients underwent AVS, 16 patients were excluded as records were unavailable. Among 97 remaining patients, 61 had unilateral disease and 57 underwent unilateral adrenalectomy. Blood pressure (BP) improved significantly with less antihypertensive medication requirement. Among those with different BP responses to adrenalectomy (cure, improvement, or no change), a higher number of preoperative antihypertensive medications was associated with persistent hypertension (P = .03). There were no significant differences in age (P = .14), duration of hypertension (P = .60), family history of hypertension (P = .68), or serum creatinine (P = .34). When AVS shows lateralization, age, duration of hypertension, family history, or renal dysfunction did not predict BP response to adrenalectomy. Results suggest that these factors should not preclude AVS and subsequent adrenalectomy. Further studies are indicated to confirm these findings.


Treatment head design for multileaf collimated high-energy electrons.
This paper describes how a conventional treatment head can be modified for use of multileaf collimated electron beams. Automatic and dynamic beam delivery are possible for both electrons and photons by using the computer controlled multileaf collimator (MLC) for both photon and electron beams. Thereby, the electron beams can be mixed more freely into the treatment to take advantage of the specific depth modulation characteristics of electrons. The investigation was based on Monte Carlo calculations using the software package BEAM. The physical parameters used in this optimization were the beam penumbra and the virtual/effective point source position. These parameters are essential for shaping beams, beam matching and for dosimetry calculations. The optimization was carried out by modifying a number of parameters: replacing the air atmosphere in the treatment head with helium, adding a helium bag below the MLC, changing the position of the scattering foils, modifying the monitor chamber, and adjusting the position of the MLC. The beam characteristics for some of these designs were found to fulfil our criteria for clinically useful beams down to at least 9 MeV.


Development and Therapeutic Potential of Small-Molecule Modulators of Circadian Systems.
Circadian timekeeping systems drive oscillatory gene expression to regulate essential cellular and physiological processes. When the systems are perturbed, pathological consequences ensue and disease risks rise. A growing number of small-molecule modulators have been reported to target circadian systems. Such small molecules, identified via high-throughput screening or derivatized from known scaffolds, have shown promise as drug candidates to improve biological timing and physiological outputs in disease models. In this review, we first briefly describe the circadian system, including the core oscillator and the cellular networks. Research progress on clock-modulating small molecules is presented, focusing on development strategies and biological efficacies. We highlight the therapeutic potential of small molecules in clock-related pathologies, including jet lag and shiftwork; various chronic diseases, particularly metabolic disease; and aging. Emerging opportunities to identify and exploit clock modulators as novel therapeutic agents are discussed.


Genetics in Idiopathic Pulmonary Fibrosis: A Clinical Perspective.
Unraveling the genetic background in a significant proportion of patients with both sporadic and familial IPF provided new insights into the pathogenic pathways of pulmonary fibrosis. The aim of the present study is to overview the clinical significance of genetics in IPF. It is fascinating to realize the so-far underestimated but dynamically increasing impact that genetics has on aspects related to the pathophysiology, accurate and early diagnosis, and treatment and prevention of this devastating disease. Genetics in IPF have contributed as no other in unchaining the disease from the dogma of a "a sporadic entity of the elderly, limited to the lungs" and allowed all scientists, but mostly clinicians, all over the world to consider its many aspects and "faces" in all age groups, including its co-existence with several extra pulmonary conditions from cutaneous albinism to bone-marrow and liver failure. By providing additional evidence for unsuspected characteristics such as immunodeficiency, impaired mucus, and surfactant and telomere maintenance that very often co-exist through the interaction of common and rare genetic variants in the same patient, genetics have created a generous and pluralistic yet unifying platform that could lead to the understanding of the injurious and pro-fibrotic effects of many seemingly unrelated extrinsic and intrinsic offending factors. The same platform constantly instructs us about our limitations as well as about the heritability, the knowledge and the wisdom that is still missing.


Enrichment and Aggregation of Purple Non-sulfur Bacteria in a Mixed-Culture Sequencing-Batch Photobioreactor for Biological Nutrient Removal From Wastewater.
Mixed-culture biotechnologies are widely used to capture nutrients from wastewater. Purple non-sulfur bacteria (PNSB), a guild of anoxygenic photomixotrophic organisms, rise interest for their ability to directly assimilate nutrients in the biomass. One challenge targets the aggregation and accumulation of PNSB biomass to separate it from the treated water. Our aim was to enrich and produce a concentrated, fast-settling PNSB biomass with high nutrient removal capacity in a 1.5-L, stirred-tank, anaerobic sequencing-batch photobioreactor (SBR). PNSB were rapidly enriched after inoculation with activated sludge at 0.1 gVSS L-1 in a first batch of 24 h under continuous irradiance of infrared (IR) light (>700 nm) at 375 W m-2, with Rhodobacter reaching 54% of amplicon sequencing read counts. SBR operations with decreasing hydraulic retention times (48 to 16 h, i.e., 1-3 cycles d-1) and increasing volumetric organic loading rates (0.2-1.3 kg COD d-1 m-3) stimulated biomass aggregation, settling, and accumulation in the system, reaching as high as 3.8 g VSS L-1. The sludge retention time (SRT) increased freely from 2.5 to 11 days. Acetate, ammonium, and orthophosphate were removed up to 96% at a rate of 1.1 kg COD d-1 m-3, 77% at 113 g N d-1 m-3, and 73% at 15 g P d-1 m-3, respectively, with COD:N:P assimilation ratio of 100:6.7:0.9 m/m/m. SBR regime shifts sequentially selected for Rhodobacter (90%) under shorter SRT and non-limiting concentration of acetate during reaction phases, for Rhodopseudomonas (70%) under longer SRT and acetate limitation during reaction, and Blastochloris (10%) under higher biomass concentrations, underlying competition for substrate and photons in the PNSB guild. With SBR operations we produced a fast-settling biomass, highly (>90%) enriched in PNSB. A high nutrient removal was achieved by biomass assimilation, reaching the European nutrient discharge limits. We opened further insights on the microbial ecology of PNSB-based processes for water resource recovery.


Human melanoma targeting with alpha-MSH-melphalan conjugate.
A conjugate made of alpha-MSH as a drug carrier and melphalan has been designed in order to target human melanoma cells. Iodination of the alpha-MSH moiety led to a relatively stable tracer which could be easily separated and analysed by reverse phase high pressure liquid chromatography. The conjugate was found to be unstable at neutral pH and a serious denaturation can take place at concentrations exceeding 100 micrograms/ml, especially in plasma. Receptor-mediated cytotoxicity has been shown by the use of cultured alpha-MSH receptor positive/negative cells as well as in vivo B16 murine melanoma model. Body distribution and uptake of the labelled compound were unaltered as compared to those of labelled free hormone. alpha-MSH receptor recognition properties also remained unchanged with a better apparent affinity of the conjugate probably due to the alkylating activity of melphalan itself. Using human melanoma dendritic cells expressing more than 10,000 alpha-MSH binding sites per cell as an in vitro model, we were able to demonstrate higher cytotoxicities as compared to melphalan-treated cells. In contrast, melanoma cells with low receptivity did not show higher cytotoxicity. P388D1 mouse plasmocytoma cells lacking receptors were much more sensitive to melphalan than the conjugate. This phenomenon appeared to be related with the number of binding sites expressed at the time of the experiment as well as cell differentiation and the doubling time. Our findings strongly support the concept of a receptor-mediated cytotoxicity and may enable the in vivo melphalan delivery to target tissues to be increased, achieving an improvement of drug penetration inside melanoma cells.


Hepatic cholesterol synthesis in man: effect of diazepam and other drugs.
The hepatic synthesis of cholesterol-measured as incorporation of 14C-acetate into digitonin-precipitable sterols-has been evaluated in needle biopsy material of normal untreated patients and patients under short-term treatment with Diazepam, Phenobarbital, Chlorpromazine or Diphenylhydantoin. A significant increase of cholesterol synthesis was observed in the first two groups with much higher levels of incorporation in the Diazepam-treated patients. Moreover in this group the levels were still elevated from 4 to 7 days after drug withdrawl. Ultrastructural and morphometric studies performed on the same biopsy material showed a significant increase of smooth endoplasmic reticulum in hepatocytes of Diazepam-treated patients; in addition, there seemed to be a positive correlation between the increased cholesterol synthesis and the formation of areas of non vesicular, type 2, smooth endoplasmic reticulum. These findings suggest an early stimulation of the liver cell microsomal system by Diazepam in man; they also point to side effects of some drugs, which are not predictable from studies in Wistar rats.


Neuroimaging-aided differential diagnosis of the depressive state.
A serious problem in psychiatric practice is the lack of specific, objective biomarker-based assessments to guide diagnosis and treatment. The use of such biomarkers could assist clinicians in establishing differential diagnosis, which may improve specific individualised treatment. This multi-site study sought to develop a clinically suitable neuroimaging-guided diagnostic support system for differential diagnosis at the single-subject level among multiple psychiatric disorders with depressive symptoms using near-infrared spectroscopy, which is a compact and portable neuroimaging method. We conducted a multi-site, case-control replication study using two cohorts, which included seven hospitals in Japan. The study included 673 patients (women/men: 315/358) with psychiatric disorders (major depressive disorder, bipolar disorder, or schizophrenia) who manifested depressive symptoms, and 1007 healthy volunteers (530/477). We measured the accuracy of the single-subject classification in differential diagnosis among major psychiatric disorders, based on spatiotemporal characteristics of fronto-temporal cortical haemodynamic response patterns induced by a brief (<3 min) verbal fluency task. Data from the initial site were used to determine an optimal threshold, based on receiver-operator characteristics analysis, and to generate the simplest and most significant algorithm, which was validated using data from the remaining six sites. The frontal haemodynamic patterns detected by the near-infrared spectroscopy method accurately distinguished between patients with major depressive disorder (74.6%) and those with the two other disorders (85.5%; bipolar disorder or schizophrenia) that presented with depressive symptoms. These results suggest that neuroimaging-guided differential diagnosis of major psychiatric disorders developed using the near-infrared spectroscopy method can be a promising biomarker that should aid in personalised care in real clinical settings. Potential confounding effects of clinical (e.g., age, sex) and systemic (e.g., autonomic nervous system indices) variables on brain signals will need to be clarified to improve classification accuracy.


Genetics of Vesicoureteral Reflux.
Vesicoureteral reflux (VUR) is the retrograde passage of urine from the bladder to the upper urinary tract. It is the most common congenital urological anomaly affecting 1-2% of children and 30-40% of patients with urinary tract infections. VUR is a major risk factor for pyelonephritic scarring and chronic renal failure in children. It is the result of a shortened intravesical ureter with an enlarged or malpositioned ureteric orifice. An ectopic embryonal ureteric budding development is implicated in the pathogenesis of VUR, which is a complex genetic developmental disorder. Many genes are involved in the ureteric budding formation and subsequently in the urinary tract and kidney development. Previous studies demonstrate an heterogeneous genetic pattern of VUR. In fact no single major locus or gene for primary VUR has been identified. It is likely that different forms of VUR with different genetic determinantes are present. Moreover genetic studies of syndromes with associated VUR have revealed several possible candidate genes involved in the pathogenesis of VUR and related urinary tract malformations. Mutations in genes essential for urinary tract morphogenesis are linked to numerous congenital syndromes, and in most of those VUR is a feature. The Authors provide an overview of the developmental processes leading to the VUR. The different genes and signaling pathways controlling the embryonal urinary tract development are analyzed. A better understanding of VUR genetic bases could improve the management of this condition in children.


Unnecessary orchiectomy due to atypical sarcoidosis manifesting as a unilateral scrotal mass: a case report and literature review.
Sarcoidosis is a disease characterised primarily by lung tissue involvement. Extrapulmonary involvement, particularly in the genitourinary tract, is extremely rare, particularly when it comes to primary disease detection in this location. The gold standard in establishing a definitive diagnosis of sarcoidosis is a combination of the clinical picture, the results of imaging methods, and histopathological examination from the biopsy taken (thus ruling out other causes of granulomatous inflammation). However, it is common for the biopsy to be infeasible or for the patient to refuse such an examination, resulting in the neglect of this critical verification. We introduce the case of a young 29-year-old man of Czech nationality who had been complaining for some time about non-specific pain above the pubic bone and in the lower abdomen, which was combined with a painless enlargement of the right half of the scrotum. Due to suspected malignancy, it was, after considering clinical, imaging, and laboratory findings, decided to perform a radical orchiectomy as a treatment option. The histological examination revealed that it was not cancer, but rather a rare genitourinary form of extrapulmonary sarcoidosis. In this case, radical resection had been, therefore, unnecessary. We also present a review of the literature on published extrapulmonary, genitourinary, and testicular sarcoidosis cases. All the above demonstrates the importance of considering a possible atypical sarcoidosis manifestation and histological confirmation before pursuing radical solutions.


Mental and personality disorders in infertile women with polycystic ovary: a case-control study.
Polycystic Ovarian Syndrome (PCOS) is one of the most common causes of infertility in women. The current study investigated mental and personality disorders in infertile women with and without PCOS. This case-control study evaluated 400 infertile women who referred to the Infertility Center in Babol city (North of Iran). Participants were categorized into the case group (201 PCOS) and the control group (199 without PCOS). All of the participants completed the Millon Clinical Multi-axial Inventory-III (MCMI-III). The mean scores for clinical personality patterns were significantly higher for six personality disorders (schizoid, avoidant, antisocial, depressive, sadistic, and negativistic) and for three classes of severe personality disorder patterns (schizotypal, borderline, and paranoid) in infertile women with PCOS than in women without PCOS. The mean scores for eight clinical disorders (somatoform, manic disorder, dysthymia, alcohol-dependence, drug-dependence, post-trauma stress disorder, major depression, and delusion disorder) were also higher in infertile women with PCOS than in women without PCOS. The scores of many mental and personality disorders are higher in infertile women with PCOS than in women without PCOS. Thus, clinicians should prioritize recognizing and treating psychological problems of infertile women with PCOS.


Actions of insulin and vitamin A on Sertoli cells.
The synthesis of androgen-binding protein by cultured Sertoli cells is increased by insulin, retinol, follitropin and testosterone. Only follitropin will stimulate an increase in cyclic AMP in these cells, yet each agent individually increased the synthesis of androgen-binding protein in short-term cultures. For long-term culture of Sertoli cells follitropin, testosterone, insulin, and retinol appeared to act synergistically to prolong the ability of the cells to secrete androgen-binding protein. These are the first reported results which suggest an action of insulin and retinol directly on Sertoli cells even though the importance of both factors in male reproduction is known.


[Development of a dissociated state in the rat after administration of carbacholine into the hippocampus].
Food-procuring conditioned reaction was elaborated in rats. A single injection of carbacholine into the dorsal area of the hippocampus elicited prolonged (for several days) stable amnesia, during which elaboration of a second habit in the same animals proved possible. After cessation of the agent action, the initially elaborated conditioned reaction recurred. The obtained results allow to interpret the amnesia elicited by injection of carbacholine into the hippocampus as a dissociated state.


Regression error estimation significantly improves the region-of-interest statistics of noisy MR images.
The region-of-interest (ROI) selection and evaluation is one of the key factors in the successful evaluation of radiological images. However, the presence of noise in images may lead to incorrect diagnosis. The aim of this study was to test the hypothesis that the weighting by error estimation in ROI assessment might significantly improve the validity of the results. As a model, the data maps of the transverse relaxation time constants (T2) from patients who underwent a matrix-associated chondrocyte transplantation procedure on the femoral condyle were analyzed. Artificial noise with a Rician density probability distribution was added to each TE image. ROIs were processed either as a regular arithmetic mean or as a weighted mean, in which weighted coefficients were calculated with regard to fitting error estimates [coefficient of determination (R2); root mean squared error (RMSE), mean absolute error (MSE), mean squared error (MAE), and chi-squared error (chi2)]. The global T2 values in repair tissue (mean +/- standard deviation, 62 +/- 7 ms; range 51-70 ms) and in healthy cartilage (mean +/- SD, 49 +/- 6 ms; range 40-60 ms) were significantly different (p < 0.001). With a 45% or greater decrease from the original SNR value (corresponding to a noise level of 35% of random value), the statistical significance was lost (P > 0.05); however, the use of the coefficient of determination (R2) as a correction factor was able to maintain the p-value of < 0.05 up to a 56% decrease from the original SNR value. The results of this study can prospectively be applied in a wide range of radiological imaging techniques in cases when error estimation is possible. Our analysis on MR images with artificially added noise showed that utilization of the correlation of determination (R2) as a weighting parameter in ROI evaluation may significantly improve the differentiation between native and transplanted cartilage tissue in noisy images. This could be an added benefit in the non-invasive monitoring of the post-operative status of patients with cartilage transplants if the MR images are not ideal (e.g., lower field strength or lower SNR).


Molecular profile of the T cell receptor beta variable in peripheral blood lymphocytes from chronic asymptomatic HBV carriers.
T cell receptor beta variable (TCRBV) repertoire could imply the composition and function status of T cells in subjects with HBV infection. The gene melting spectral pattern (GMSP) can be used to determine the profile of TCRBV gene family. The molecular profile of TCRBV in peripheral lymphocytes from asymptomatic HBV carriers (AsC) remains ill-defined. Peripheral blood mononuclear cells (PBMCs) were separated and sorted, and the profiles of TCRBV complementarity-determining region 3 (CDR3) in CD4(+), CD8(+) T subsets and PBMCs were assayed using GMSP. The number of skewed TCRBV in the PBMCs was significantly lower than that in the CD4(+) or CD8(+) T subsets, and the number of monoclonal TCRBV families in the CD8(+) T subset was significantly higher than that in CD4(+) T subset. Compared to healthy donors, TCRBV11, BV13.1, BV20 and BV24 were used more frequently than other TCRBV members in PBMCs from AsC subjects. Furthermore, the relatively conserved CDR3 motifs were detected in these TCRBVs. The results indicate that the T cell response in AsC subjects involves several TCRBVs, and that the CD8(+) T subset maybe more relevant to pathogenesis of AsC. Moreover, the four relative conserved TCRBVs maybe a target for personalized treatments for persistent HBV infection.


Coencapsulation of butyl-methoxydibenzoylmethane and octocrylene into lipid nanocarriers: UV performance, photostability and in vitro release.
The coencapsulation of two UV filters, butyl-methoxydibenzoylmethane (BMDBM) and octocrylene (OCT), into lipid nanocarriers was explored to develop stable cosmetic formulations with broad-spectrum photoprotection and slow release properties. Different types of nanocarriers in various concentrations of the two UV filters were tested to find the combination with the best absorption and release properties. Solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs) have been the two types of lipid nanocarriers used. The NLCs were based on either medium chain triglycerides (MCT) or squalene (Sq). The following physicochemical properties of the nanocarriers have been evaluated: particle size, morphology, zeta potential (ZP), entrapment efficiency, loading capacity, and thermal behavior. The nanocarriers have been formulated into creams containing low amounts of UV filters (2.5% BMDBM and 1% OCT). The best photoprotection results were obtained with the cream based on NLCs prepared with MCT, having a sun protection factor (SPF) of 17.2 and an erythemal UVA protection factor (EUVA-PF) of 50.8. The photostability of the encapsulated BMDBM filter was confirmed by subjecting the nanocarriers-based creams to in vitro irradiation. The prolonged UV-protection efficacy was coupled with a slow in vitro release of the synthetic UV filters, which followed the Higuchi release model.


Genital findings in adolescent girls referred for suspected sexual abuse.
Sexual abuse is a common problem affecting adolescent girls, but the frequency of medical findings in this population has not been specifically described. To describe the frequency of specific genital findings in a group of pubertal girls who had experienced probable or definite sexual abuse. Patient series, medical chart and photograph review. Specialty referral clinic for abused children. PATIENTS AND SELECTION: Referred sample of female patients, examined between January 1, 1987, and June 30, 1994, with Tanner genital stages 3, 4, or 5, who reported a history of penile-vaginal penetration, had colposcopic photographs taken, and were determined, by means of a previously described classification system, to have experienced probable or definite abuse. None. The study included 204 girls, aged 9 to 17 years (mean, 13 years); race or ethnicity was Mexican American in 57%, white in 34%, and other in 9%. Abnormal genital findings were documented in 32% of patients overall but were more common when the girls had reported bleeding at the time of the assault (50% vs 26%; P = .004, chi 2 analysis), or when the examination occurred within 72 hours of the last episode of abuse (69% vs 26%; P < .001, chi 2 analysis). Transections of the hymen were unusual (8%), but notches in the hymen were more common (25%). Normal or nonspecific results of genital examinations are commonly found in adolescents who have been sexually abused, unless the abuse was very recent. Further studies are needed to document the healing of genital injuries in victims of acute assault and the frequency of hymenal findings in nonabused, non-sexually active adolescents.


Presence of follipsin complexed with alpha 2-macroglobulin in porcine ovarian follicular fluid.
Using the follicular fluid of porcine ovaries as a source, a high-molecular-weight protein having enzyme activity toward t-butyloxycarbonyl-Gln-Arg-Arg-4-methylcoumaryl-7-amide was purified to an apparent homogeneity. Its molecular weight was estimated to be approximately 720,000. The protein was immunoprecipitated with antihuman alpha 2-macroglobulin antibody and was visualized in Western blot analysis using the same antibody. Using Western blot analysis, an 85 kDa polypeptide, which was recognized by antiporcine follipsin antibodies, was detected in this high-molecular-weight protein. When tested for substrates and inhibitors with low molecular weight, the protein showed substrate specificity and an inhibitor profile similar to those of follipsin. On the basis of these results, it was concluded that the protein is a complex of follipsin with alpha 2-macroglobulin. The complexed enzyme activated a single-chain precursor tissue-type plasminogen activator at a drastically reduced rate compared with free follipsin. The present results suggest that intrafollicular follipsin activity is regulated by the proteinase inhibitor alpha 2-macroglobulin.


Recovery of a function involving gene duplication by retroposition in Saccharomyces cerevisiae.
The duplication of DNA sequences contributes to genomic plasticity and is known to be one of the key factors responsible for evolution. The mechanisms underlying these rare events, which have been frequently mentioned by authors performing genomic analysis, have not yet been completely elucidated. These mechanisms were approached here in the yeast Saccharomyces cerevisiae, using a positive selection screen based on a particular mutated allele of the URA2 gene. Spontaneous revertants containing a duplication of the terminal part of the URA2 gene were selected and analyzed. Some important features of the duplicated regions, such as their chromosome location, size, and insertion sites, were characterized. The events selected correspond to a single inter- or intrachromosomal gene duplication process. The duplicated ATCase sequence is generally punctuated by a poly(A) tract and is always located in Ty1 sequences. In addition, the activation of a Ty1 transcription process increased the frequency of the duplication events. All in all, these data suggest that the duplication mechanism involves the reverse transcription of mRNA and the subsequent integration of the cDNA into a Ty1 area. The Ty1 elements and the retrotransposon-encoded function are key factors contributing to chromosomal reshaping. The genomic rearrangements described constitute experimental evidence for the recovery of a function involving duplication by retroposition.


Machine Learning Approach to Decision Making for Insulin Initiation in Japanese Patients With Type 2 Diabetes (JDDM 58): Model Development and Validation Study.
Applications of machine learning for the early detection of diseases for which a clear-cut diagnostic gold standard exists have been evaluated. However, little is known about the usefulness of machine learning approaches in the decision-making process for decisions such as insulin initiation by diabetes specialists for which no absolute standards exist in clinical settings. The objectives of this study were to examine the ability of machine learning models to predict insulin initiation by specialists and whether the machine learning approach could support decision making by general physicians for insulin initiation in patients with type 2 diabetes. Data from patients prescribed hypoglycemic agents from December 2009 to March 2015 were extracted from diabetes specialists' registries, resulting in a sample size of 4860 patients who had received initial monotherapy with either insulin (n=293) or noninsulin (n=4567). Neural network output was insulin initiation ranging from 0 to 1 with a cutoff of >0.5 for the dichotomous classification. Accuracy, recall, and area under the receiver operating characteristic curve (AUC) were calculated to compare the ability of machine learning models to make decisions regarding insulin initiation to the decision-making ability of logistic regression and general physicians. By comparing the decision-making ability of machine learning and logistic regression to that of general physicians, 7 cases were chosen based on patient information as the gold standard based on the agreement of 8 of the 9 specialists. The AUCs, accuracy, and recall of logistic regression were higher than those of machine learning (AUCs of 0.89-0.90 for logistic regression versus 0.67-0.74 for machine learning). When the examination was limited to cases receiving insulin, discrimination by machine learning was similar to that of logistic regression analysis (recall of 0.05-0.68 for logistic regression versus 0.11-0.52 for machine learning). Accuracies of logistic regression, a machine learning model (downsampling ratio of 1:8), and general physicians were 0.80, 0.70, and 0.66, respectively, for 43 randomly selected cases. For the 7 gold standard cases, the accuracies of logistic regression and the machine learning model were 1.00 and 0.86, respectively, with a downsampling ratio of 1:8, which were higher than the accuracy of general physicians (ie, 0.43). Although we found no superior performance of machine learning over logistic regression, machine learning had higher accuracy in prediction of insulin initiation than general physicians, defined by diabetes specialists' choice of the gold standard. Further study is needed before the use of machine learning-based decision support systems for insulin initiation can be incorporated into clinical practice.


Virtual reality for treatment compliance for people with serious mental illness.
Virtual reality (VR) is computerised real-time technology, which can be used an alternative assessment and treatment tool in the mental health field. Virtual reality may take different forms to simulate real-life activities and support treatment. To investigate the effects of virtual reality to support treatment compliance in people with serious mental illness. We searched the Cochrane Schizophrenia Group Trials Register (most recent, 17th September 2013) and relevant reference lists. All relevant randomised studies comparing virtual reality with standard care for those with serious mental illnesses. We defined virtual reality as a computerised real-time technology using graphics, sound and other sensory input, which creates the interactive computer-mediated world as a therapeutic tool. All review authors independently selected studies and extracted data. For homogeneous dichotomous data the risk difference (RD) and the 95% confidence intervals (CI) were calculated on an intention-to-treat basis. For continuous data, we calculated mean differences (MD). We assessed risk of bias and created a 'Summary of findings' table using the GRADE approach. We identified three short-term trials (total of 156 participants, duration five to 12 weeks). Outcomes were prone to at least a moderate risk of overestimating positive effects. We found that virtual reality had little effects regarding compliance (3 RCTs, n = 156, RD loss to follow-up 0.02 CI -0.08 to 0.12, low quality evidence), cognitive functioning (1 RCT, n = 27, MD average score on Cognistat 4.67 CI -1.76 to 11.10, low quality evidence), social skills (1 RCT, n = 64, MD average score on social problem solving SPSI-R (Social Problem Solving Inventory - Revised) -2.30 CI -8.13 to 3.53, low quality evidence), or acceptability of intervention (2 RCTs, n = 92, RD 0.05 CI -0.09 to 0.19, low quality evidence). There were no data reported on mental state, insight, behaviour, quality of life, costs, service utilisation, or adverse effects. Satisfaction with treatment - measured using an un-referenced scale - and reported as "interest in training" was better for the virtual reality group (1 RCT, n = 64, MD 6.00 CI 1.39 to 10.61,low quality evidence). There is no clear good quality evidence for or against using virtual reality for treatment compliance among people with serious mental illness. If virtual reality is used, the experimental nature of the intervention should be clearly explained. High-quality studies should be undertaken in this area to explore any effects of this novel intervention and variations of approach.


SARS-CoV-2 replicon for high-throughput antiviral screening.
The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) virus, which is highly pathogenic and classified as a biosafety level 3 (BSL-3) agent, has greatly threatened global health and efficacious antivirals are urgently needed. The high requirement of facilities to manipulate the live virus has limited the development of antiviral study. Here, we constructed a reporter replicon of SARS-CoV-2, which can be handled in a BSL-2 laboratory. The Renilla luciferase activity effectively reflected the transcription and replication levels of the replicon genome. We identified the suitability of the replicon in antiviral screening using the known inhibitors, and thus established the replicon-based high-throughput screening (HTS) assay for SARS-CoV-2. The application of the HTS assay was further validated using a few hit natural compounds, which were screened out in a SARS-CoV-2 induced cytopathic-effect-based HTS assay in our previous study. This replicon-based HTS assay will be a safe platform for SARS-CoV-2 antiviral screening in a BSL-2 laboratory without the live virus.


Changes in speed distribution: Applying aggregated safety effect models to individual vehicle speeds.
This study investigated the effect of applying two aggregated models (the Power model and the Exponential model) to individual vehicle speeds instead of mean speeds. This is of particular interest when the measure introduced affects different parts of the speed distribution differently. The aim was to examine how the estimated overall risk was affected when assuming the models are valid on an individual vehicle level. Speed data from two applications of speed measurements were used in the study: an evaluation of movable speed cameras and a national evaluation of new speed limits in Sweden. The results showed that when applied on individual vehicle speed level compared with aggregated level, there was essentially no difference between these for the Power model in the case of injury accidents. However, for fatalities the difference was greater, especially for roads with new cameras where those driving fastest reduced their speed the most. For the case with new speed limits, the individual approach estimated a somewhat smaller effect, reflecting that changes in the 15th percentile (P15) were somewhat larger than changes in P85 in this case. For the Exponential model there was also a clear, although small, difference between applying the model to mean speed changes and individual vehicle speed changes when speed cameras were used. This applied both for injury accidents and fatalities. There were also larger effects for the Exponential model than for the Power model, especially for injury accidents. In conclusion, applying the Power or Exponential model to individual vehicle speeds is an alternative that provides reasonable results in relation to the original Power and Exponential models, but more research is needed to clarify the shape of the individual risk curve. It is not surprising that the impact on severe traffic crashes was larger in situations where those driving fastest reduced their speed the most. Further investigations on use of the Power and/or the Exponential model at individual vehicle level would require more data on the individual level from a range of international studies.


[Segmental instability and lumbar spinal canal stenosis. Theoretical, clinical and surgical aspects].
Clinical, radiological, surgical and experimental observations make it possible to construct a rational theory of the segmental lumbar instability which explains the pathogenesis of spondylosis, of stenosis and of several cases of disc displacement as well. This theory helps us to choose the right surgical procedure. Spinal stenosis, degenerative spondylolisthesis and some cases of disc displacement are seen as part of a degenerative process with its starting point in regressive changes of the intervertebral disc and lateral joint. This leads to a marked instability of the affected mobile segment. Signs and symptoms of unstable lumbar spine are discussed. In such cases, if decompression of the roots by laminectomy, facetectomy or discectomy is performed without fusion, the consequences are usually treated but not the main cause of the trouble, namely the instability. Finally, we describe a procedure which we prefer as the safest and best of all, using selected decompression by means of joint screws (spondylodesis) as suggested by F. Magerl (41-44).


Acyclovir therapy of varicella-zoster virus infections in immunocompromised patients.
In a randomized, placebo-controlled, double-blind trial of intravenous acyclovir in the treatment of varicella zoster virus (VZV) infections, 8 of 20 immunocompromised children with varicella received acyclovir (500 mg/m2/dose three times daily for 7 days). There was no significant difference in skin healing between the acyclovir and placebo groups although there was a significant reduction in the incidence of development of pulmonary involvement during acyclovir treatment. Nineteen out of 34 patients received vidarabine (10 mg/kg/day for 5 days). Vidarabine significantly shortened the duration of new vesicle formation. Both drugs significantly reduced the incidence of visceral varicella, the most serious complication of VZV infection. An open trial also concluded that early treatment of varicella in these patients is essential. Of the 94 patients with zoster infection, 52 received acyclovir (500 mg/m2/dose infused over one hour three times daily for 7 days). Acyclovir recipients healed more rapidly, had fewer days of pain and shorter duration of viral shedding compared with placebo patients. The most important finding was that acyclovir significantly protected against progression of zoster as defined by development or progression of cutaneous dissemination and development of visceral zoster. Vidarabine seemed to be equally effective in this respect. The likelihood of cutaneous dissemination is related to the nature of the underlying condition. The in vitro sensitivity of VZV isolates from patients with second episode VZV infection during the trial did not change appreciably which suggests that VZV does not become resistant to acyclovir during therapy.


The prevalence of traumatic brain injury among young offenders in custody: a systematic review.
To examine the prevalence of traumatic brain injury (TBI) among young people in custody and to compare this with estimates within the general youth population. Systematic review of research from various national contexts. Included studies were assessed for the relevance of the definition of TBI and the research population, and the quality of the study design. Ten studies were identified for inclusion in the review. Four of these studies included control groups. No studies examining comorbidity of TBI and other neurodevelopmental disorders among incarcerated young people were identified. Reported prevalence rates of brain injury among incarcerated youth range from 16.5% to 72.1%, with a rate of 100% reported among a sample of young people sentenced to death. This suggests considerable levels of need among incarcerated young people. Where control groups or directly comparable studies within the general population exist, there is strong and consistent evidence of a prevalence of TBI among incarcerated youth that is substantially greater than that in the general population. This disparity is seemingly more pronounced as the severity of the injury increases.


Lecithin-cholesterol acyltransferase (LCAT) activity in chronic uremia.
High plasma concentrations of triglycerides and low plasma concentrations of esterified cholesterol and lysolecithin, with an impaired rate of VLDL and LDL catabolism, have been reported in chronic uremic patients. An important contribution to these abnormalitites might be an impaired activity of the (LCAT). Serum LCAT activity and cholesteryl ester clearance were determined in 11 patients with chronic renal failure and in 10 controls. LCAT activity was determined by using the serum of each patient both as a source of enzyme and as a substrate ("intrinsic" activity) and was compared with the activity determined on a standard substrate ("extrinsic activity), so as to ascertain the presence of inhibitory factors in the patients' sera. Both activityes have been found to be significantly (P less than 0.01) lower in chronic uremic patients than in controls. The cholesteryl ester clearance apparently did not respond to the stimulatory effect of hypertriglyceridemia, as observed in other cases of dislipoproteinemias. The parallel decrease of both enzyme activities makes it unlikely that it is due to the presence of "uremic toxins" inhibiting the enzyme activity. LCAT synthesis in the liver is probably reduced in chronic uremia. These results suggest that in chronic uremia the VLDL fail to cooperate in their own catabolism.


Psychopharmacogenetic basis of medication-induced movement disorders.
In light of the emerging evidence for genetic vulnerability to adverse drug reactions, this article attempts to elucidate the natural history of medication-induced movement disorders from a psychopharmacogenetic perspective. Studies of the risk factors, neurobiology, and pharmacogenetics are reviewed concurrently. The relevant variables associated with 10 genetically mediated movement disorders are tabulated and compared with those of medication-induced movement disorders without a clear-cut genetic basis. As a result of this integrated analysis, it is evident that there is an intimate genetic and pathophysiological link between neuropsychiatric movement disorders of diverse origins. The emergence of drug-induced movement disorders seems to reflect a spectrum of basal ganglia derangement attributable to genetic predisposition; psychotropic medications only augment the genetic vulnerability to clinical phenotypes. It is proposed that a multidimensional analysis of the interacting variables is essential for understanding the natural history of these conditions, and that the scope of psychopharmacology should be broadened to include psychopharmacogenetics for improving therapeutic objectivity and prevention research.


HeLa cell response proteome alterations induced by mammalian reovirus T3D infection.
Cells are exposed to multiple stressors that induce significant alterations in signaling pathways and in the cellular state. As obligate parasites, all viruses require host cell material and machinery for replication. Virus infection is a major stressor leading to numerous induced modifications. Previous gene array studies have measured infected cellular transcriptomes. More recently, mass spectrometry-based quantitative and comparative assays have been used to complement such studies by examining virus-induced alterations in the cellular proteome. We used SILAC (stable isotope labeling with amino acids in cell culture), a non-biased quantitative proteomic labeling technique, combined with 2-D HPLC/mass spectrometry and reciprocal labeling to identify and measure relative quantitative differences in HeLa cell proteins in purified cytosolic and nuclear fractions after reovirus serotype 3 Dearing infection. Protein regulation was determined by z-score analysis of each protein's label distribution. A total of 2856 cellular proteins were identified in cytosolic fractions by 2 or more peptides at >99% confidence and 884 proteins were identified in nuclear fractions. Gene ontology analyses indicated up-regulated host proteins were associated with defense responses, immune responses, macromolecular binding, regulation of immune effector processes, and responses to virus, whereas down-regulated proteins were involved in cell death, macromolecular catabolic processes, and tissue development. These analyses identified numerous host proteins significantly affected by reovirus T3D infection. These proteins map to numerous inflammatory and innate immune pathways, and provide the starting point for more detailed kinetic studies and delineation of virus-modulated host signaling pathways.


DNA sequence recognition by bleomycin.
The ability of bleomycin to cleave DNA fragments containing (TA)n and (GA)n/(CT)n inserts has been investigated. Bleomycin cuts efficiently after AT and GA though not at TA, AG and CT or TC. In the presence of ligands which modify DNA structure new cleavage products appear. It seems that the ability of bleomycin to discriminate between various sequences arises from their structural characteristics rather than merely from base-specific hydrogen bond contacts.


The response of streams in the Adirondack region of New York to projected changes in sulfur and nitrogen deposition under changing climate.
Modeling studies project that in the future surface waters in the northeast US will continue to recover from acidification over decades following reductions in atmospheric sulfur dioxide and nitrogen oxides emissions. However, these studies generally assume stationary climatic conditions over the simulation period and ignore the linkages between soil and surface water recovery from acid deposition and changing climate, despite fundamental impacts to watershed processes and comparable time scales for both phenomena. In this study, the integrated biogeochemical model PnET-BGC was applied to two montane forest watersheds in the Adirondack region of New York, USA to evaluate the recovery of surface waters from historical acidification in response to possible future changes in climate and atmospheric sulfur and nitrogen deposition. Statistically downscaled climate scenarios on average project warmer temperatures and greater precipitation for the Adirondack by the end of the century. Model simulations suggest under constant climate, acid-sensitive Buck Creek would gain 12.8 μeq L-1 of acid neutralizing capacity (ANC) by 2100 from large reductions in deposition, whereas acid insensitive Archer Creek is projected to gain 7.9 μeq L-1 of ANC. However, climate change could limit those improvements in acid-base status. Under climate change, a negative offset relative to the ANC increases with no climate change are projected for both streams by 2100. In acid-insensitive Archer Creek the negative offset (-8.5 μeq L-1) was large enough that ANC is projected to decrease by -0.6 μeq L-1, whereas in acid-sensitive Buck Creek, the negative offset (-0.4 μeq L-1) resulted in a slight decline of the projected future ANC increase to 12.4 μeq L-1. Calculated target loads for 2150 for both sites decreased when future climate change was considered in model simulations, which suggests further reductions in acid deposition may be necessary to restore ecosystem structure and function under a changing climate.


Final report of the amended safety assessment of myristic acid and its salts and esters as used in cosmetics.
This report addresses the safety of the inorganic salts and esters of various fatty alcohols of myristic acid. Most of the esters are used as skin conditioning agents in many types of cosmetics in a range of concentrations. Myristate esters are readily hydrolyzed to the corresponding alcohols and acids, which are then further metabolized. Myristate salts readily dissociate in any likely cosmetic formulation. The Cosmetic Ingredient Review (CIR) Panel recognized that much of the data supporting the ingredients in this group were previously reviewed in safety assessments for related ingredients. Where specific data did not exist, the Panel considered structure-activity relationships in determining the safety of these ingredients as used in cosmetics. The Panel determined that myristic acid and its salts and esters are safe as cosmetic ingredients in the current practices of use and concentration.


Lead the way, Leader.
Effective leaders show objectivity, creativity, and knowledge. They develop personal leadership styles from existing models and use their strength to motivate staff, accomplish organizational goals, and develop future leaders.


Serial changes in lymphocyte subsets in patients with newly diagnosed high grade astrocytomas treated with standard radiation and temozolomide.
The immune system plays a significant role in cancer prevention and outcome. In high grade astrocytomas (HGA), severe lymphopenia is associated with shortened survival due to tumor progression. This study was performed to quantify serial changes in lymphocyte subsets in HGA following standard radiation (RT) and temozolomide (TMZ). Adults (KPS >60, HIV negative) with newly diagnosed HGA scheduled to receive concurrent RT and TMZ and adjuvant TMZ were eligible. Blood was collected before beginning concurrent RT/TMZ and at weeks 6, 10, 18, and 26, and 3 months after completing adjuvant TMZ. Lymphocyte subsets were analyzed by flow cytometry. Twenty patients (70% glioblastoma, median age 53, 50% male, 80% Caucasian) who enrolled from January 2014 to August 2014 were followed until April 2016. Baseline dexamethasone dose was 0.5 mg/day and 15% had absolute lymphocyte counts (ALC) <1000 cells/mm3 before starting RT/TMZ. However, 75% developed lymphopenia with ALC <1000 cells/mm3 after completion of RT/TMZ. NK cells, B cells and all T lymphocytes subsets dropped significantly after concurrent RT/TMZ and remained depressed for the 48 weeks of observation. The CD4+/CD8+ ratio was not affected significantly during follow-up. Severe lymphopenia involving all subsets occurred early in treatment and remained present for nearly 1 year. To our knowledge, this is the first report of serial trends in lymphocyte subsets following standard RT and TMZ for HGA.


Prolonged end-stage fetal heart rate deceleration: a reanalysis.
In an earlier publication we reported on 18 patients who had what was described as prolonged fetal heart rate (FHR) deceleration in the second stage of labor and who had uniformly good outcome. It was believed that the reason for the good outcome was early diagnosis and rapid delivery when the FHR deceleration did not improve by the time the patient entered the delivery room. We have continued to handle this type of deceleration pattern in a similar fashion with good results, but 18 additional cases of prolonged end-stage FHR deceleration associated with a poor outcome are reported along with 1 case of intrauterine demise. Physicians who use the fetal monitor need to be aware of this potentially ominous FHR pattern.


Brunner syndrome associated MAOA mutations result in NMDAR hyperfunction and increased network activity in human dopaminergic neurons.
Monoamine neurotransmitter abundance affects motor control, emotion, and cognitive function and is regulated by monoamine oxidases. Among these, Monoamine oxidase A (MAOA) catalyzes the degradation of dopamine, norepinephrine, and serotonin into their inactive metabolites. Loss-of-function mutations in the X-linked MAOA gene have been associated with Brunner syndrome, which is characterized by various forms of impulsivity, maladaptive externalizing behavior, and mild intellectual disability. Impaired MAOA activity in individuals with Brunner syndrome results in bioamine aberration, but it is currently unknown how this affects neuronal function, specifically in dopaminergic (DA) neurons. Here we generated human induced pluripotent stem cell (hiPSC)-derived DA neurons from three individuals with Brunner syndrome carrying different mutations and characterized neuronal properties at the single cell and neuronal network level in vitro. DA neurons of Brunner syndrome patients showed reduced synaptic density but exhibited hyperactive network activity. Intrinsic functional properties and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR)-mediated synaptic transmission were not affected in DA neurons of individuals with Brunner syndrome. Instead, we show that the neuronal network hyperactivity is mediated by upregulation of the GRIN2A and GRIN2B subunits of the N-methyl-d-aspartate receptor (NMDAR), resulting in increased NMDAR-mediated currents. By correcting a MAOA missense mutation with CRISPR/Cas9 genome editing we normalized GRIN2A and GRIN2B expression, NMDAR function and neuronal population activity to control levels. Our data suggest that MAOA mutations in Brunner syndrome increase the activity of dopaminergic neurons through upregulation of NMDAR function, which may contribute to the etiology of Brunner syndrome associated phenotypes.


Knowledge and attitudes to personal genomics testing for complex diseases among Nigerians.
The study examined the knowledge and attitudes to personal genomics testing for complex diseases among Nigerians and identified how the knowledge and attitudes vary with gender, age, religion, education and related factors. Data were collected using qualitative method in 2 districts of the Federal Capital Territory. In the study, eight (8) Focused Group Discussions (FGDs) and twenty seven (27) Key Informant Interviews (KIIs) were conducted. Participants for the research were recruited among healthy Nigerians, individuals with complex diseases, health care professionals, community leaders and health policy makers. Analysis of the result showed that most respondents in both FGDs and KIIs had limited knowledge about genomics test initially. Their understanding of the test however improved after explanation on its concept. Participants showed positive attitude towards genomics tests. Nevertheless they expressed fear over direct to consumer personal genomics testing, testing unborn babies and disclosure of results to third parties. Culture and religion were found to influence the perspectives of respondents on genomics test particularly those aspects that could either directly contradict their beliefs and practices or lead to actions which contradict them. In conclusion, most Nigerians interviewed had limited knowledge of genomics test but with supportive attitude towards its use in predicting future risk of complex diseases after understanding the test concept. Genomics testing for complex diseases was not a common practice in Nigeria.


Electrically evoked responses in onset chopper neurons in guinea pig cochlear nucleus.
Extracellular recordings were obtained from single cochlear nucleus neurons in guinea pigs anesthetized with Nembutal and Hypnorm. Neurons were classified by their spontaneous firing rates and responses to acoustic stimuli. In addition, electrical shocks were applied to the midline at the level of the IVth ventricle and spike responses were recorded. Spikes were evoked by shocks only in neurons that were classified as onset choppers (O(c)). The shock-evoked spikes could be extinguished by acoustically evoked action potentials in the same neurons. In roughly 30% of the sample of O(c) neurons, quantitative aspects of the timing of this extinction were not compatible with the shock-evoked spike being antidromically conducted from O(c) output axons. Together with the presence of temporal jitter at high shock rates, the data suggest the possibility that at least some of the shock-evoked spikes may be generated by excitatory synaptic input to the O(c) neurons, most likely from the collaterals of the medial olivocochlear system (MOCS), whose axons pass close to the floor of the IVth ventricle. This excitatory synaptic input may operate to modulate the activity of O(c) neurons in addition to MOCS actions in the auditory periphery.


Facultative anoxygenic photosynthesis in the cyanobacterium Oscillatoria limnetica.
An isolate from H2S-rich layers of the Solar Lake, the cyanobacterium Oscillatoria limnetica, exhibits both oxygenic and anoxygenic photosynthesis. It can use Na2S as an electron donor for CO2 photoassimilation (photosystem I supplies the energy) in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea or 700-nm light. A stoichiometric ratio of approximately 2 is observed between the Na2S consumed and the photoassimilated CO2. The anoxygenic phototrophic capability of this cyanobacterium explains its growth in nature in high sulfide concentrations and indicates a selective advantage.


[The significance of goniometric data for the estimation of the harm to health inflicted to the victims of ankle joint injuries].
The present paper is devoted to the forensic medical estimation of the harm to health inflicted to the victims of ankle joint injuries with reference to the role and significance of traditional goniometry in the practical work of traumatologists, orthopedists and forensic medical experts. It is shown that the use of goniometric data alone as the sole criterion for the estimation of the harm to health in case of ankle joint injuries is unjustified and hampers further development of this field of forensic medicine.


Mixture models and wavelet transforms reveal high confidence RNA-protein interaction sites in MOV10 PAR-CLIP data.
The Photo-Activatable Ribonucleoside-enhanced CrossLinking and ImmunoPrecipitation (PAR-CLIP) method was recently developed for global identification of RNAs interacting with proteins. The strength of this versatile method results from induction of specific T to C transitions at sites of interaction. However, current analytical tools do not distinguish between non-experimentally and experimentally induced transitions. Furthermore, geometric properties at potential binding sites are not taken into account. To surmount these shortcomings, we developed a two-step algorithm consisting of a non-parametric two-component mixture model and a wavelet-based peak calling procedure. Our algorithm can reduce the number of false positives up to 24% thereby identifying high confidence interaction sites. We successfully employed this approach in conjunction with a modified PAR-CLIP protocol to study the functional role of nuclear Moloney leukemia virus 10, a putative RNA helicase interacting with Argonaute2 and Polycomb. Our method, available as the R package wavClusteR, is generally applicable to any substitution-based inference problem in genomics.


Use of prescription medication in the last years of life: a population-based comparison of two oldest old Danish birth cohorts born 10 years apart.
Age limits for diagnostics and treatments have been largely removed and replaced by an active diagnostic and treatment practice among the oldest old and has led to concerns about potential overtreatment during the last years of life. Use of prescription medication in the last years of life was assessed from 1995 to 2012 for the entire 1905 and 1915 Danish birth cohorts using nationwide register data. Medication use was quantified as the number different pharmacy-redeemed drugs during 120 days up to a given date. For both cohorts, prescription medication use increased with proximity to death and calendar year, while age at death had little impact; use in the 1915 cohort was markedly higher than in the 1905 cohort. Average number of prescription medications varied from below 3 to above 9 depending on age, calendar year and proximity to death. From 1995 to 2005, average number of prescription medications for a 90-year-old person in the last month of life increased from 6.0 to 8.7. Out of 90-year-old persons dying in 2005, 82% were exposed to polypharmacy, up from 63% in 1995. Prescription medication use accelerates throughout the last of years life among two Danish oldest old cohorts born 10 years apart, with substantially larger use in the most recent cohort. This pattern suggests an increase in drug prescribing regimens in the period 1995-2012, reinforcing the need for evidence-based guidelines on medications in the particularly vulnerable population of the oldest old patients in their last years of life.


Hydrophilic stationary phases: a practical approach for the co-analysis of compounds with varying polarity in biological matrices.
The aim was to simultaneously extract, separate and detect not only the opioid methadone and its primary metabolites (2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine and 2-ethyl-5-methyl-3,3-diphenyl-1-pyrroline), but also creatinine from urine, plasma and fingerprints. Creatinine is highly polar and analysis by RP chromatography using conventional stationary phases such as silica-bonded C8 and C18 is unsuitable. Hydrophilic stationary phases are increasingly being applied for the analysis of highly polar analytes, this chemistry being investigated as a suitable alternative. A hydrophilic interaction liquid chromatography phase column successfully retained creatinine and permitted the co-analysis of methadone and its metabolites by LC-MS/MS. Prior to analysis, an extraction protocol for urine and plasma was required but for fingerprint deposits this was not necessary. Alteration of sample pH, necessary to extract methadone, and its metabolites led to difficulties associated with the extraction of creatinine. This problem was addressed by first performing an SPE incorporating a hydrophilic interaction phase to extract creatinine, and the eluent then combined with the opioid extract from a mixed-mode cation exchange phase. The assay for creatinine, methadone and its primary phase I metabolites met validation criteria. LC-MS/MS analysis of creatinine and drug compounds together offers considerable advantages over traditional approaches that necessitate the quantification of creatinine using spectrophotometric approaches.


Generalized provisional seed zones for native plants.
Deploying well-adapted and ecologically appropriate plant materials is a core component of successful restoration projects. We have developed generalized provisional seed zones that can be applied to any plant species in the United States to help guide seed movement. These seed zones are based on the intersection of high-resolution climatic data for winter minimum temperature and aridity (as measured by annual heat : moisture index), each classified into discrete bands. This results in the delineation of 64 provisional seed zones for the continental United States. These zones represent areas of relative climatic similarity, and movement of seed within these zones should help to minimize maladaptation. Superimposing Omernik's level III ecoregions over these seed zones distinguishes areas that are similar climatically yet different ecologically. A quantitative comparison of provisional seed zones with level III ecoregions and provisional seed zones within ecoregions for three species showed that provisional seed zone within ecoregion often explained the greatest proportion of variation in a suite of traits potentially related to plant fitness. These provisional seed zones can be considered a starting point for guidelines for seed transfer, and should be utilized in conjunction with appropriate species-specific information as well as local knowledge of microsite differences.


Designing a Care Pathway Model - A Case Study of the Outpatient Total Hip Arthroplasty Care Pathway.
Although the clinical attributes of total hip arthroplasty (THA) care pathways have been thoroughly researched, a detailed understanding of the equally important organisational attributes is still lacking. The aim of this article is to contribute with a model of the outpatient THA care pathway that depicts how the care team should be organised to enable patient discharge on the day of surgery. The outpatient THA care pathway enables patients to be discharged on the day of surgery, shortening the length of stay and intensifying the provision and organisation of care. We utilise visual care modelling to construct a visual design of the organisation of the care pathway. An embedded case study was conducted of the outpatient THA care pathway at a teaching hospital in the Netherlands. The data were collected using a visual care modelling toolkit in 16 semi-structured interviews. Problems and inefficiencies in the care pathway were identified and addressed in the iterative design process. The results are two visual models of the most critical phases of the outpatient THA care pathway: diagnosis & preparation (1) and mobilisation & discharge (4). The results show the care team composition, critical value exchanges, and sequence that enable patient discharge on the day of surgery. The design addressed existing problems and is an optimisation of the case hospital's pathway. The network of actors consists of the patient (1), radiologist (1), anaesthetist (1), nurse specialist (1), pharmacist (1), orthopaedic surgeon (1,4), physiotherapist (1,4), nurse (4), doctor (4) and patient application (1,4). The critical value exchanges include patient preparation (mental and practical), patient education, aligned care team, efficient sequence of value exchanges, early patient mobilisation, flexible availability of the physiotherapist, functional discharge criteria, joint decision making and availability of the care team.


Expression of PD-1 and PD-L1 in Extramammary Paget Disease: Implications for Immune-Targeted Therapy.
Extramammary Paget disease (EMPD) is a locally aggressive cutaneous malignancy that usually arises in anogenital or axillary skin. Immune checkpoint inhibitors targeting programmed cell death receptor (PD-1) and/or its ligand (PD-L1) are approved for the treatment of several types of cancer, and response to these generally correlates with increased PD-L1 expression by tumor cells. The expression of PD-L1 and composition and density of the tumor-associated immune infiltrate in EMPD have been little studied. To determine whether EMPD might be amenable to immune checkpoint blockade, we analyzed the expression of PD-1 and PD-L1 and the composition and density of the tumor-associated immune infiltrate in EMPD and evaluated associations between biomarker expression and clinicopathologic parameters. Twenty-one EMPD tumors were evaluated for tumor cell PD-L1 expression and for relative expression and distribution of CD3, CD8, PD-1, and PD-L1 in the tumor-associated immune infiltrate by using a combination of visual and image analysis (Aperio ImageScope). In addition, PD-L1 expression was assessed in 10 cases of mammary Paget disease (MPD). In EMPD cases, PD-L1 was expressed by tumor cells (3/21; 14%) and the tumor-associated immune infiltrate (15/21; 71%), and PD-1 was expressed by the tumor-associated immune infiltrate in all cases analyzed (18/18). However, PD-L1 expression by EMPD tumor cells did not correlate with the density of CD3-, CD8-, or PD-1-positive cells in the tumor-associated immune infiltrate or other clinicopathologic parameters. Furthermore, the density of CD3, CD8, PD-1, and PD-L1 in the tumor-associated immune infiltrate did not correlate with any clinicopathologic parameters evaluated with the exception that CD3 positive values were significantly higher in patients who were still alive (median, 1310 cells/mm2; range, 543-2115;) than in those who died (median, 611 cells/mm2; range, 481-908; p = 0.049). In all MPD cases, PD-L1 was absent in tumor cells but present in the tumor-associated immune infiltrate, and PD-L1 expression in lymphocytes was lower in patients with HER2/neu-positive than in those with HER2/neu-negative disease (p = 0.07). Our findings raise the possibility of therapeutic targeting of the PD-1/PD-L1 axis in EMPD.


[Embolism in the right heart chambers: the diagnostic and therapeutic aspects].
Deep venus thrombosis may result in pulmonary embolism. In rare instances, embolization has occurred, not directly to the pulmonary arterial tree, but to the right heart chambers. Although the value of echocardiography in the diagnosis is well recognised, their is no consensus for the appropriate treatment. We report herein six cases of floating right atrial thrombi, diagnosed by echocardiography, in patients with pulmonary embolism, or unexplained shock or syncope. Surgical embolectomy was carried out in 4 patients, and thrombolytic therapy in 2, without in-hospital mortality. The high mortality associated to this entity may be improved by rapid echocardiographic recognition and emergency treatment with thrombolysis or surgery. Our data suggest the possible use of thrombolysis as a first-choice therapy in selected patients.


Differential regulation of the cell wall integrity mitogen-activated protein kinase pathway in budding yeast by the protein tyrosine phosphatases Ptp2 and Ptp3.
Mitogen-activated protein kinases (MAPKs) are inactivated by dual-specificity and protein tyrosine phosphatases (PTPs) in yeasts. In Saccharomyces cerevisiae, two PTPs, Ptp2 and Ptp3, inactivate the MAPKs, Hog1 and Fus3, with different specificities. To further examine the functions and substrate specificities of Ptp2 and Ptp3, we tested whether they could inactivate a third MAPK, Mpk1, in the cell wall integrity pathway. In vivo and in vitro evidence indicates that both PTPs inactivate Mpk1, but Ptp2 is the more effective negative regulator. Multicopy expression of PTP2, but not PTP3, suppressed growth defects due to the MEK kinase mutation, BCK1-20, and the MEK mutation, MKK1-386, that hyperactivate this pathway. In addition, deletion of PTP2, but not PTP3, exacerbated growth defects due to MKK1-386. Other evidence supported a role for Ptp3 in this pathway. Expression of MKK1-386 was lethal in the ptp2Delta ptp3Delta strain but not in either single PTP deletion strain. In addition, the ptp2Delta ptp3Delta strain showed higher levels of heat stress-induced Mpk1-phosphotyrosine than the wild-type strain or strains lacking either PTP. The PTPs also showed differences in vitro. Ptp2 was more efficient than Ptp3 at binding and dephosphorylating Mpk1. Another factor that may contribute to the greater effectiveness of Ptp2 is its subcellular localization. Ptp2 is predominantly nuclear whereas Ptp3 is cytoplasmic, suggesting that active Mpk1 is present in the nucleus. Last, PTP2 but not PTP3 transcript increased in response to heat shock in a Mpk1-dependent manner, suggesting that Ptp2 acts in a negative feedback loop to inactivate Mpk1.


Overexpression of XIAP expression in renal cell carcinoma predicts a worse prognosis.
X-linked inhibitor of apoptosis protein (XIAP) is the most potent caspase-inhibitory IAP family member and a negative regulator of various apoptotic stimuli. Thus, XIAP overexpression in cancer cells may select for tumor cell survival following various cytotoxic therapeutic modalities. The anatomical staging system in renal cell carcinoma (RCC) currently provides good prognostic information, albeit insufficient. We hypothesize that overexpression of XIAP in RCC may serve as a molecular prognostic marker in RCC and improve the staging of RCC. This study examined the protein level of XIAP in lysates from surgical specimens of 109 patients with RCC and 109 normal kidney specimens from the same patients. The level of XIAP expression was quantified by Western blot analysis using non-fixed fresh frozen tissues of RCCs and normal kidneys. Results indicated that the mean level of XIAP expression was higher in RCC compared to autologous normal kidney, and the XIAP expression level in 38/109 (35%) of RCC was more than 2-fold greater than that in normal kidney tissue. In Stage I/II RCC, the mean XIAP expression level was almost identical to that detected in normal kidney, whereas XIAP expression in Stage III/IV was 2.5-fold higher than that in Stage I/II RCC. Levels of XIAP expression also correlated with the grade of RCC. Patients with RCC with low XIAP expression had a longer postoperative disease-specific survival as compared to those with high expression in the 5-year follow-up. The suggested role of XIAP in the regulation of resistance in apoptosis was examined in vitro following treatment of RCC cell lines with XIAP antisense oligonucleotide and the cells were sensitized to both Fas-mediated and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis. The present study demonstrates at the protein level that XIAP is overexpressed in RCC, and that high XIAP expression in RCC predicted a worse prognosis. In addition, XIAP antisense oligonucleotide sensitized RCC to Fas/TRAIL-induced apoptosis. These results suggest that XIAP expression in RCC may be used as a prognostic parameter, and that downregulation or inhibition of XIAP expression in RCC may reverse immune resistance.


System influences on posthomicide beliefs and distress.
Criminal justice system experiences in 150 family members of homicide victims were investigated. The study had two goals: (a) to document the experiences of homicide survivors in the criminal justice system, including case outcomes, criminal justice system activities, and satisfaction with system personnel; and (b) to determine if experiences with the police impacted posthomicide beliefs (cognitive assumptions about the world and one's relationship to it) and psychological distress. The sample, which was identified through the Medical Examiner's Office, was drawn from all criminal homicides from 1.5 to 5 years prior to selection. Results showed that family members of homicide victims were very dissatisfied with their experiences in the criminal justice system. Additionally, whereas objective system outcomes (e.g., arrest) did not directly affect posthomicide beliefs and distress, subjective system outcomes (e.g., police satisfaction) directly affected beliefs and indirectly affected distress. There was some support for both equity theory and a cognitive theory of change, the two theories that guided the model specification.


[Urogenital myiasis with Fannia scalaris].
A 15-year-old boy excreted a fly larva in his urine. It constituted the third larval state of the fly Fannia scalaris. Microhaematuria, mild albuminuria and leucocyturia ceased after the parasite had been passed.


Disruption of electrostatic contacts in the HNH nuclease from a thermophilic Cas9 rewires allosteric motions and enhances high-temperature DNA cleavage.
Allosteric signaling within multidomain proteins is a driver of communication between spatially distant functional sites. Understanding the mechanism of allosteric coupling in large multidomain proteins is the most promising route to achieving spatial and temporal control of the system. The recent explosion of CRISPR-Cas9 applications in molecular biology and medicine has created a need to understand how the atomic level protein dynamics of Cas9, which are the driving force of its allosteric crosstalk, influence its biophysical characteristics. In this study, we used a synergistic approach of nuclear magnetic resonance (NMR) and computation to pinpoint an allosteric hotspot in the HNH domain of the thermostable GeoCas9. We show that mutation of K597 to alanine disrupts a salt-bridge network, which in turn alters the structure, the timescale of allosteric motions, and the thermostability of the GeoHNH domain. This homologous lysine-to-alanine mutation in the extensively studied mesophilic S. pyogenes Cas9 similarly alters the dynamics of the SpHNH domain. We have previously demonstrated that the alteration of allostery via mutations is a source for the specificity enhancement of SpCas9 (eSpCas9). Hence, this may also be true in GeoCas9.


Coronal suturectomy does not cause acute postoperative displacement in the cranial bases of craniosynostotic rabbits.
Approximately 300 to 500 infants per 1,000,000 have prematurely fused cranial sutures (craniosynostosis). Craniosynostosis can result in increased intracranial pressure and craniofacial deformities, which often require extensive and costly craniofacial surgery. Because the neurocranium and basicranium are developmentally interrelated, understanding their influence on one another is important for surgical planning. Although surgery has been found to have long-term effects on the cranial bases of craniosynostotic human beings and rabbits, the biological mechanisms behind these effects remain uncertain. Some researchers have suggested that the surgical release of synostosed sutures alters long-term growth patterns, resulting in cranial base differences between craniosynostotic individuals who undergo surgery and those who do not. Additionally, some investigators have proposed that an acute and surgically related displacement of basicranial elements may contribute to the observed differences. The current study examines acute postoperative changes in four lengths, three angles, and three triangles between cranial base landmarks in a sample of seven New Zealand white rabbits with familial nonsyndromic craniosynostosis. Results indicate that suturectomy caused no statistically significant (P < 0.05) acute length, angular, or shape differences in the cranial base. Thus, previous long-term cranial base differences found between rabbits that were operated on and those that were not were probably not caused by an acute displacement of skeletal elements as a result of surgery. These findings suggest that postoperative cranial base changes may be related more to chronically altered growth patterns than to acutely altered changes in intracranial pressure or dural tension.


Use of fluorescence spectroscopy for quantitative investigations of ubiquitin interactions with the ubiquitin-binding domains of NEMO.
Ubiquitin serves as a signal for a variety of cellular processes and its specific interaction with ubiquitin-binding domain (UBD) regulates key cellular events including protein degradation, cell-cycle control, DNA repair, and kinase activation. Several binding mechanisms for isolated UBDs have been reported in recent years. However, little is known about the mechanism through which proteins containing multiple-UBDs achieve specificity for a particular oligomer of polyUb. The NF-κB essential modulator (NEMO, also known IKKγ), which plays a key role in the NF-κB signaling pathway, belongs to the latter family of proteins since it contains two distal NOA (also known UBAN/CC2-LZ/NUB) and ZF UBDs, separated by an unstructured proline-rich linker of about 40 residues in length. Here, we show a new procedure for fast purification of this bipartite domain. We also describe the use of intrinsic fluorescence spectroscopy for quantitative investigations of ubiquitin interactions between two distal ubiquitin-binding domains of NEMO (NOA and ZF). This spectroscopic method has many advantages over other techniques like GST pulldown and Biacore's SPR for monitoring avid interactions between two UBDs, especially when UBDs are located at significant distance from each other within the protein.


Hemobilia in systemic lupus erythematosus.
Symptoms of acute cholecystitis developed in a 22-year-old woman with active SLE. Abdominal ultrasonography and biliary patency scan showed evidence of acalculous cholecystitis and common bile duct obstruction, respectively. Operation revealed acalculous cholecystitis and hemobilia; a liver biopsy specimen also showed hemobilia. Surgery relieved the patient's symptoms. This case demonstrates a new complication of SLE.


Opportunities for healthcare digitalization in Europe: Comparative analysis of inequalities in access to medical services.
Digitalization of healthcare systems is a great opportunity to address inequalities in access to healthcare in the European Union. There is an urgent need to build on what we learned from the COVID-19 pandemic, where digital health technologies were integrated swiftly to limit challenges in healthcare delivery. We created a database for the 27 European Union countries from the European Health Interview Survey (EHIS), the Digital Economy and Society Index (DESI), and other Eurostat databases. We performed k-means cluster analysis to group EU countries along two dimensions: inequalities in access to medical services and level of digitalization. We identified five distinct clusters: two clusters with high, two clusters with moderate, and one cluster with low unmet need for healthcare. Regarding digitalization, only one cluster comprising the Nordic countries, Spain and Cyprus exhibit high digital readiness. A cluster comprising the most developed countries in Western Europe represents moderate levels of both unmet need for healthcare and digitalization. For most EU countries, there is still a need to build digital infrastructure for the healthcare industry, which in the long term may increase the number of digital solutions used by both patients and healthcare professionals. Policy makers across the EU need to consider investing in initiatives that would support digital health solutions as an effective means of healthcare provision and healthcare management.


Optimization of operating room allocation using linear programming techniques.
New and innovative approaches must be used to rationally allocate scarce resources such as operating room time while simultaneously optimizing the associated financial return. In this article we use the technique of linear programming to optimize allocation of OR time among a group of surgeons based on professional fee generation. For the period of December 1, 2000, to July 31, 2002, the following individualized data were obtained for the Division of General Surgery at Duke University Medical Center: allocated OR time (hours), case mix as determined by CPT codes, total OR time used, and normalized professional charges and receipts. Inpatient, outpatient, and emergency cases were included. The Solver linear programming routine in Microsoft Excel (Microsoft Corp.) was used to determine the optimal mix of surgical OR time allocation to maximize professional receipts. Our model of optimized OR allocation would maximize weekly professional revenues at 237,523 US dollars, a potential increase of 15% over the historical value of 207,700 US dollars or an annualized increase of approximately 1.5 million US dollars. Our results suggest that mathematical modeling techniques used in operations research, management science, or decision science may rationally optimize OR allocation to maximize revenue or to minimize costs. These techniques may optimize allocation of scarce resources in the context of the goals specific to individual academic departments of surgery.


Retrieval of precursors for white-type adipose conversion in brown adipose tissue.
A cellular compartment from brown adipose tissue (BAT) of newborn rats was isolated by Percoll-density-gradient centrifugation and was shown to proliferate and to undergo adipose conversion in vitro in primary culture. The features of the effector requirement for adipose conversion as well as the differentiated morphological and biochemical phenotype are almost identical with that of a compartment designated HCF, from white adipose tissue (WAT). A possible role for these precursors from BAT and WAT in the involution of BAT into WAT, on the one hand, and in the development of brown adipose cells among typical WAT deposits, on the other, is discussed.


Sleep behavior and chronotype before and throughout pregnancy.
To compare sleep behavior before and during pregnancy. In this prospective cohort study, healthy women were followed from pre-pregnancy until delivery. At pre-pregnancy and each trimester, participants completed validated questionnaires of chronotype and sleep quality and timing, including the Munich ChronoType Questionnaire, Epworth Sleepiness Scale, and Pittsburgh Sleep Quality Index. The primary outcomes were sleep period start and end times, sleep duration, sleep midpoint, and social jetlag, compared between pre-pregnancy and each trimester. Wrist actigraphy was used to measure the same outcomes in a subset of participants. Eighty-six women were included in analysis of questionnaires. Of these, 37 provided complete actigraphy data. Questionnaire and actigraphy data indicate that participants had less social jetlag during pregnancy than before pregnancy. Sleep period start times were earlier on both work and free days in the first and second trimesters than pre-pregnancy, and returned to pre-pregnancy times by the third trimester. Actigraphy data revealed that, compared to pre-pregnancy, participants had longer sleep periods in all trimesters on work days and in the first trimester on free days. Sleep surveys revealed that participants had poorer sleep quality in the first and third trimesters and more sleepiness in the first trimester than pre-pregnancy. The first trimester of pregnancy is characterized by earlier sleep period start time, longer sleep duration, and poorer sleep quality than pre-pregnancy. Sleep quality temporarily improves in the second trimester, and sleep period start time returns to pre-pregnancy time by the third trimester. Multiple parameters of sleep have been studied in the context of pregnancy and pregnancy outcomes, but rarely in comparison to pre-pregnancy or longitudinally through pregnancy. Actigraphy and questionnaire data reveal sleep timing and quality change throughout pregnancy. These data on sleep changes in healthy pregnancy can be used as a baseline to identify sleep-related risk factors throughout pregnancy.