The management and treatment of an abnormal Pap smear.
Table 2 summarizes the management of the abnormal Pap smear. Management of dysplasia in this institution is aggressive--as destructive therapy of mild dysplasia is advised, opposed to watching the patient and treating only if the disease persists. The rationale for this is the 33% to 45% failure rate for follow-up appointments in the primarily inner-city population served. The key to follow-up is to repeat cervical cytology in all patients treated, even those treated with hysterectomy, every three months until two consecutive normal smears are obtained. At that time, surveillance and intervals may be modified, but screening should continue at least annually. The mortality rate of carcinoma of the cervix has dropped precipitously during the last 40 years, in part, from simple screening of the cervix with the Papanicolaou smear. The effort to treat premalignant changes has been rewarded. The use of the colposcope and destructive forms of therapy have allowed successful treatment of patients with less morbidity and mortality than the immediate reliance on cervical conization. Remember, conization is still indicated and prudent in selected patients. Following these guidelines may contribute to the downward trend.


Evaluation of a chromogenic agar (MRSASelect) for the detection of meticillin-resistant Staphylococcus aureus with clinical samples in The Netherlands.
A novel chromogenic medium for the detection of meticillin-resistant Staphylococcus aureus (MRSA), MRSASelect (Bio-Rad), was evaluated with clinical samples in a public health laboratory in The Netherlands. In total, 3000 samples were tested in the period January to March 2005, including 972 nose, 972 throat, 968 perineum, and 88 wound or urine samples. Presumptive MRSA colonies appeared pink/mauve on the MRSASelect medium. The performance of MRSASelect medium was compared with the routine screening method. Evaluation of the colony morphology showed that all confirmed MRSA isolates grew as pink/mauve colonies. None of the white colonies were MRSA strains. The number of false-positive pink/mauve colonies increased after prolonged incubation from 20 to 48 h. The specificity decreased from 92 % after 20 h incubation to 89 % after 48 h incubation. In total 70 MRSA strains were isolated, 55 of which were detected by the MRSASelect medium and 55 were detected by the routine screening method. Sensitivity was 78.6 % for both test procedures, and specificities were 99.5 and 100 %, respectively for the MRSASelect medium and the routine screening method. The addition of an enrichment broth to the MRSASelect medium increased the number of MRSA strains detected by 12 %. In total, 18 patients were MRSA positive, 4 of these were detected by the MRSASelect medium only and 1 was detected by the routine screening method only. Sensitivity on patient level was 94.4 and 77.8 % for the MRSASelect medium and the routine screening method, respectively, while specificities were 99.7 and 99.0 %.


[Digestive hemorrhage caused by duodenal varices].
Duodenal varices are uncommon and are frequently diagnosed late. However, they are a potential cause of massive bleeding, with tendency to relapse. We would like to offer as an example a case diagnosed by means of an endoscopy. A patient without oesophageal varices and with obstruction of the splenic-portal axis and venous calcifications is reported. A diminution in the volume of the duodenal varices with inspiratory movements may help in the differential diagnosis during endoscopy. We emphasize the need of avoiding incorrect maneuvers during endoscopy and the value of ultrasonography and CAT scan to confirm the diagnosis. Sclerotherapy appears to be the best therapeutic approach. Percutaneous embolization and surgery are therapeutic alternatives if sclerosis is not effective. However, in some cases with self-limited bleeding and a low tendency to relapse, as with the patient described herein, a conservative attitude should be selected.


Assessing soil properties and nutrient availability under conservation agriculture practices in a reclaimed sodic soil in cereal-based systems of North-West India.
Soil quality degradation associated with resources scarcity is the major concern for the sustainability of conventional rice-wheat system in South Asia. Replacement of conventional management practices with conservation agriculture (CA) is required to improve soil quality. A field experiment was conducted to assess the effect of CA on soil physical (bulk density, penetration resistance, infiltration) and chemical (N, P, K, S, micronutrients) properties after 4 years in North-West India. There were four scenarios (Sc) namely conventional rice-wheat cropping system (Sc1); partial CA-based rice-wheat-mungbean system (RWMS) (Sc2); CA-based RWMS (Sc3); and CA-based maize-wheat-mungbean (Sc4) system. Sc2 (1.52 Mg m-3) showed significantly lower soil bulk density (BD). In Sc3 and Sc4, soil penetration resistance (SPR) was reduced and infiltration was improved compared to Sc1. Soil organic C was significantly higher in Sc4 than Sc1. Available N was 33% and 68% higher at 0-15 cm depth in Sc3 and Sc4, respectively, than Sc1. DTPA extractable Zn and Mn were significantly higher under Sc3 and Sc4 compared to Sc1. Omission study showed 30% saving in N and 50% in K in wheat after four years. Therefore, CA improved soil properties and nutrient availability and have potential to reduce external fertilizer inputs in long run.


Effects of Escherichia coli and Bacteroides fragilis on peritoneal host defenses.
Escherichia coli and Bacteroides fragilis are common copathogens in clinical intra-abdominal sepsis, yet it is unclear how they interact synergistically in vivo. We sought to determine whether E. coli and B. fragilis, in combination but not alone, could exert a detrimental effect on the peritoneal host defenses of translymphatic absorption and bacterial phagocytosis. Our data indicated that nonviable E. coli (O18ab:K56/K7:- and O111:B4), Klebsiella pneumoniae, B. fragilis, and Bacteroides thetaiotaomicron were handled in a similar fashion by both host defenses of the peritoneal cavity. The use of 2 X 10(8) nonviable radiolabeled E. coli as a tracer and either 2 X 10(9) B. fragilis or 2 X 10(9) E. coli (either viable or nonviable) as a competing agent to inhibit host defenses demonstrated that although clearance and phagocytosis could be inhibited, the inhibition occurred to a similar degree with either E. coli or B. fragilis. Thus, B. fragilis did not compete to any greater extent than E. coli did for peritoneal clearance or opsonization and phagocytosis in vivo. These data indicate that bacterial synergy probably does not occur on the basis of reduced peritoneal clearance or by a reduction in the opsonization and phagocytosis of either organism by the copathogen. These results provide indirect support for the hypothesis that in bacterial synergy, one organism directly stimulates the growth of the other, perhaps by providing a growth factor.


[Intraoperative echography in biliary tract surgery. A comparative study of intraoperative echography/cholangiography in the detection of calculi in the common bile duct].
We report 3 years of experience with operative ultrasonography on 200 patients who underwent cholecystectomy for biliary lithiasis. Intraoperative ultrasonography and intraoperative cholangiography were performed on all of these patients and compared with the operative findings. The diagnostic accuracy of sonography was 98% and that of cholangiography 96% in the whole patient group. The predictive value of a positive ultrasonography was 92.5% while that of a positive cholangiogram was lower at 81.5%. In 32 patients who underwent surgical exploration of the common duct the use of these two intraoperative screening tests together led to a positive common duct exploration in 75% of the patients. Overall morbidity in this series was 4.8% against 12.8% (common duct exploration) and retained stones following duct exploration were present in 1 patient in intra-hepatic situation. The overall mortality rate of the entire patient group was 0%. Operative ultrasonography of the biliary tract as a screening procedure is a reliable method, a possible substitute for operative cholangiography, but perhaps in reality both methods are complementary.


The effects of frequency response on speech perception for cochlear implant users.
The aim of this study was to investigate the effects on speech perception of manipulating filter gains in a cochlear implant speech processor. Five implantees, who use the CI22 implant and Spectra processor manufactured by Cochlear Ltd, participated. Four experimental maps were created that were identical to their clinical map except for the profile of gains across the filters. Experimental gain profiles had rising or falling gains across the frequency range, or emphasized or de-emphasized the middle frequencies, relative to the clinical map. Perception of CNC (consonant-vowel-consonant) words at 70 dB SPL was significantly better with the clinical map than with all experimental maps, whereas at the lower level (60 dB SPL) there was minimal difference between the maps, with the low-frequency emphasis map giving significantly better scores than the high-frequency emphasis map. Perception of sentences at 70 dB SPL with a signal-to-noise ratio of +10 dB was better with the high-frequency emphasis map than with the low-frequency emphasis map. None of these best-conditions, however, were statistically better than the clinical map. The results highlighted the importance of signal audibility for speech perception with cochlear implants.


Adaptive optics optical coherence tomography in glaucoma.
Since the introduction of commercial optical coherence tomography (OCT) systems, the ophthalmic imaging modality has rapidly expanded and it has since changed the paradigm of visualization of the retina and revolutionized the management and diagnosis of neuro-retinal diseases, including glaucoma. OCT remains a dynamic and evolving imaging modality, growing from time-domain OCT to the improved spectral-domain OCT, adapting novel image analysis and processing methods, and onto the newer swept-source OCT and the implementation of adaptive optics (AO) into OCT. The incorporation of AO into ophthalmic imaging modalities has enhanced OCT by improving image resolution and quality, particularly in the posterior segment of the eye. Although OCT previously captured in-vivo cross-sectional images with unparalleled high resolution in the axial direction, monochromatic aberrations of the eye limit transverse or lateral resolution to about 15-20 μm and reduce overall image quality. In pairing AO technology with OCT, it is now possible to obtain diffraction-limited resolution images of the optic nerve head and retina in three-dimensions, increasing resolution down to a theoretical 3 μm3. It is now possible to visualize discrete structures within the posterior eye, such as photoreceptors, retinal nerve fiber layer bundles, the lamina cribrosa, and other structures relevant to glaucoma. Despite its limitations and barriers to widespread commercialization, the expanding role of AO in OCT is propelling this technology into clinical trials and onto becoming an invaluable modality in the clinician's arsenal.


Host impact of monogenean Lepidotrema bidyana infection and intensity estimates for onsite monitoring.
We developed a rapid effective method for accurate estimation of intensity for the monogenean Lepidotrema bidyana, a gill parasite of silver perch Bidyanus bidyanus. This parasite requires monitoring because high-intensity infections reduce host growth and can lead to secondary bacterial and fungal infections. The most accurate method for counting L. bidyana was visual examination of fresh gills. There was a significant relationship between fish size and parasite intensity; however, there was no significant relationship between fish condition and parasite intensity. Parasite intensity estimates were generated by using the mean intensity of worms on the posterior hemibranch on the first left gill arch, compared to the total mean intensity of worms on all hemibranchs. Estimates were validated by predicting L. bidyana intensity from a random sample of silver perch obtained from aquaculture ponds. Parasite intensity estimates correlated strongly to real counts, and this method can be used to accurately predict parasite intensity on an individual host, and thus represents an improvement over previous methods.


Physicochemical Investigation into Major League Baseballs in the Era of Unprecedented Rise in Home Runs.
A major attraction of baseball is the home run. Throughout baseball's history, some seasonal increases in home run numbers have been tied to external influences, such as lowering the pitching mound or a change in the ball manufacturer. In Major League Baseball, a recent surge in home runs has led to speculation about the baseball being "juiced" or altered in a way to make it fly farther. To support multiple academic and journalistic studies, which have attempted to find evidence of the changes in the flight of the baseball, a systematic chemical analysis has been reported on the multicomponent baseballs. Thus, we undertook a study where we analyzed the core of the baseball using various chemical and physical techniques. Studies using computed tomography scans revealed that there is a drastic 56.7% difference in the density of the core of the baseballs used during the 2014 and pre-All-Star Game 2015 versus 2017 season in the Major League. Increased material porosity was observed using electron microscopy. Thermogravimetric and elemental analyses of the pill material showed a 7% difference in the ratio of organic to inorganic material and almost a 10% decrease in silicon, respectively. Overall, the data indicates a difference in the core of the baseballs between the two time periods, leaving the contribution of these measured differences in the explosion of home runs open to interpretation.


Patient safety instruction in US health professions education.
To describe patient safety instruction in health professional curricula, including medicine, nursing, pharmacy, and dentistry. A systematic review of the literature from 1966 through 2010 was conducted using 6 databases and based on 3 search criteria: safety management, patient safety, and curriculum. One hundred fifty-four articles were identified and 23 met inclusion criteria. A variety of educational methods have been used in health profession curricula to promote patient safety including lectures, workshops, objective structured clinical examinations, standardized patients, simulation exercises, root cause analysis, quality assurance projects, and other interactive learning methods. The development of patient safety curricula has been primarily discipline-specific, with little interdisciplinary research found. Safe, patient-centered care is directly influenced by the quality of education that healthcare professions students receive. From this literature review, research is needed to guide curricular change, specifically focusing on instructional methods and interdisciplinary collaborations.


Quantitative evaluation of interstitial fibrosis with Sirius Red in IgA nephritis.
Tubulointerstitial fibrosis is one of the strongest independent predictive factors in determining the prognosis in IgA nephritis. Recently, software-based quantitative measurement of interstitial fibrosis with Sirius Red staining has entered the practice. The objective of this study was to investigate the prognostic value of measurement of interstitial nephritis with this method in IgA nephritis. Forty-three patients diagnosed with IgA nephritis with renal biopsy between the years 2005 and 2009 were included in this retrospective observational study. The diagnostic biopsies of 37 patients were examined. Basal data included age, gender, creatinine level, glomerular filtration rate (GFR), presence of proteinuria, hypertension, glomerulosclerosis, mesangial proliferation, and interstitial fibrosis and fibrosis index calculated by the measurement of computed images of Sirius Red positive areas. Final visit included evaluation of development of end-stage renal disease (ESRD), and GFR (whether = 60 mL/min or <60 mL/min). Numbers of patients with hypertension (75% vs. 34.5%; p = 0.050), ESRD development (62.5% vs. 20.7%, p = 0.035), GFR <60 mL/min (87.5% vs. 31%; p = 0.007) were greater; and basal GFR (34.25 ± 25.29 vs. 64.14 ± 35.34; p = 0.048) was lower in high-intensity interstitial fibrosis group (>1000 μm2) compared to low-intensity interstitial fibrosis group (≤1000 μm(2)). Quantitative analysis of computed imaging of areas of Sirius Red positive tubulointerstitial fibrosis might serve as an effective novel method to determine the prognosis in IgA nephritis.


MOCCA: Multilayer One-Class Classification for Anomaly Detection.
Anomalies are ubiquitous in all scientific fields and can express an unexpected event due to incomplete knowledge about the data distribution or an unknown process that suddenly comes into play and distorts the observations. Usually, due to such events' rarity, to train deep learning (DL) models on the anomaly detection (AD) task, scientists only rely on "normal" data, i.e., nonanomalous samples. Thus, letting the neural network infer the distribution beneath the input data. In such a context, we propose a novel framework, named multilayer one-class classification (MOCCA), to train and test DL models on the AD task. Specifically, we applied our approach to autoencoders. A key novelty in our work stems from the explicit optimization of the intermediate representations for the task at hand. Indeed, differently from commonly used approaches that consider a neural network as a single computational block, i.e., using the output of the last layer only, MOCCA explicitly leverages the multilayer structure of deep architectures. Each layer's feature space is optimized for AD during training, while in the test phase, the deep representations extracted from the trained layers are combined to detect anomalies. With MOCCA, we split the training process into two steps. First, the autoencoder is trained on the reconstruction task only. Then, we only retain the encoder tasked with minimizing the L2 distance between the output representation and a reference point, the anomaly-free training data centroid, at each considered layer. Subsequently, we combine the deep features extracted at the various trained layers of the encoder model to detect anomalies at inference time. To assess the performance of the models trained with MOCCA, we conduct extensive experiments on publicly available datasets, namely CIFAR10, MVTec AD, and ShanghaiTech. We show that our proposed method reaches comparable or superior performance to state-of-the-art approaches available in the literature. Finally, we provide a model analysis to give insights regarding the benefits of our training procedure.


Clinical and Echocardiographic Risk Factors Predict Late Recurrence after Radiofrequency Catheter Ablation of Atrial Fibrillation.
The benefits of radiofrequency catheter ablation (RFCA) for patients with atrial fibrillation (AF) significantly decrease with late recurrence (LR). We aimed to develop a scoring system to identify patients at high and low risk for LR following RFCA, based on a comprehensive evaluation of multiple risk factors for AF recurrence, including echocardiographic parameters. We studied 2,352 patients with AF undergoing first-time RFCA in a single institution. The LR-free survival rate up to 5 years was measured using a Kaplan-Meier analysis. The influence of clinical and echocardiographic parameters on LR was calculated with a Cox-regression analysis. Duration of AF ≥4 years (hazard ratio [HR] = 1.75; p < 0.001), non-paroxysmal AF (HR = 3.18; p < 0.001), and diabetes (HR = 1.34; p = 0.015) were associated with increased risk of LR. Left atrial (LA) diameter ≥45 mm (HR = 2.42; p < 0.001), E/e' ≥ 10 (HR = 1.44; p < 0.001), dense SEC (HR = 3.30; p < 0.001), and decreased LA appendage flow velocity (≤40 cm/sec) (HR = 2.35; p < 0.001) were echocardiographic parameters associated with increased risk of LR following RFCA. The LR score based on the aforementioned risk factors could be used to predict LR (area under curve = 0.717) and to stratify the risk of LR (HR = 1.45 per 1 point increase in the score; p < 0.001). In conclusion, LR after RFCA is affected by multiple clinical and echocardiographic parameters. This study suggests that combining these multiple risk factors enables the identification of patients with AF at high or low risk for having arrhythmia recurrence.


Clinical and Financial Implications of Positive Margins After Nonmelanoma Skin Cancer Resection: A Longitudinal Evaluation.
Management of positive margins after nonmelanoma skin cancer (NMSC) excision is debated in the literature. The purpose of this study is to determine the rate of residual tumor in reexcised NMSC specimens after previous excision with positive margins, to determine the rate of recurrence in patients who had positive margins but did not undergo reexcision, and to define the financial burden of negative reexcisions. An Institutional Review Board-approved retrospective review was conducted on all patients with NMSC excision over a 15-year period. Patients who met inclusion criteria and underwent initial NMSC excision with negative frozen section margins, but had positive permanent section margins were divided into 2 groups: those who underwent reexcision for clearance of tumor (n = 161) or those who did not undergo further reexcision (n = 105). Variables collected include demographics, previous skin cancer, tumor location, cancer subtype, excision measurements, and time between first and second excisions. For those patients who did not undergo reexcision, charts were examined for recurrence. Two hundred sixty-six patients met inclusion criteria with mean follow-up of 60 months. Eighty-three (52%) of 161 patients with positive margins on initial excision had no evidence of residual cancer upon reexcision. Residual tumor on permanent section was confirmed in 48% of patients. Patients with a previous history of basal cell carcinoma were more likely to have a true-positive margin after reexcision (P = 0.02). Larger reexcisions were more likely to harbor residual cancer (5.9 cm2, P = 0.04). Patients with positive margins that did not undergo reexcision, only 7 of 105 patients (6.6%) had recurrence. No mortalities were reported from NMSC recurrence. US $247,672 was spent in reexcision for negative margins in 98 patients for an average cost of US $2984 per case. Forty-eight percent of NMSC patients with positive margins had residual tumor upon reexcision. There were 6.6% of the patients who did not undergo resection after positive margins developed recurrence of disease at 5 years. Patients requiring larger reexcisions or those with a prior history of BCC were more likely to have residual cancer upon reexcision. This study suggests that observation is an appropriate option of care for certain patients with residual NMSC on permanent pathology.


The Relationships Between Chocolate Consumption and Endothelial Dysfunction in Patients with Heart Failure.
Dietary recommendations, in addition to medications, have recently become important in the treatment of heart failure. Our study aimed to show the positive effects of both milk chocolate and dark chocolate on heart failure through endothelial functions. Twenty patients with heart failure and reduced ejection fraction were included in the study. In this randomized, crossover study, some of the patients consumed milk chocolate and some consumed dark chocolate. We recorded the patients' 6-minute walking tests, flow- mediated dilatation values, plasma catechin, epicatechin, and N-terminal pro-brain natri- uretic peptide values before and after chocolate consumption. After 2 weeks, their chocolate consumption was changed. The same parameters were measured again. A significant decrease was observed in N-terminal pro-brain natriuretic peptide values after consumption of both milk chocolate (356 ± 54.2 and 310 ± 72.1 pg/mL; P = .007) and dark chocolate (341 ± 57 and 301 ± 60.1 pg/mL;P=.028). Flow-mediated dilation values increased after dark chocolate consumption (8.9 ± 3% and 14 ± 4.5%; P = .019). Chocolate consumption acutely decreases N-terminal pro-brain natriuretic pep- tide values in heart failure. Dark chocolate consumption also seems to improve endothelial functions by increasing flow-mediated dilation values.


Neuroimaging in nine patients with inversion duplication of the short arm of chromosome 8.
Inversion duplication 8p is a rare chromosome disorder characterised by severe mental retardation, minor facial dysmorphisms, and corpus callosum agenesis. Few cerebral imaging studies have been reported. We describe 9 patients with an inversion duplication 8p, involving variable segments of the short arm of chromosome 8. MRI was performed in 8 patients and 2 patients underwent CT scanning. The images were systematically reviewed. All patients suffered from severe mental retardation. Head circumference was between 0 and + 2 SD in 7 patients. The corpus callosum was absent in 6, and thin (but complete) in 3 patients. Hypoplasia of the (inferior) cerebellar vermis was observed in 6 patients. Enlargement of the ventricular system and associated hippocampal maldevelopment were found in all patients. The supratentorial external CSF spaces were enlarged in 6 patients, and in 3 patients there was a remarkable enlargement of the retrocerebellar arachnoidal space. Cerebral white matter showed mildly delayed myelination in 7 patients, and periventricular lesions of variable extent in 6 patients. The pattern of imaging abnormalities was non-specific, but remarkably similar between patients. We found no correlation between the severity of the clinical features, imaging results, and extent of the chromosomal aberration.


AIM1, a novel non-lens member of the betagamma-crystallin superfamily, is associated with the control of tumorigenicity in human malignant melanoma.
AIM1 is a novel gene whose expression is associated with the experimental reversal of tumorigenicity of human malignant melanoma. The predicted protein product of the major 4.1-kb transcript shows striking similarity to the betagamma-crystallin superfamily. All known members of this superfamily contain two or four characteristic motifs arranged as one or two symmetrical domains. AIM1, in contrast, contains 12 betagamma motifs, suggesting a 6-domain structure resembling a trimer of beta- or gamma-crystallin subunits. The structure of the AIM1 gene shows remarkable similarity to beta-crystallin genes, with homologous introns delineating equivalent protein structural units. AIM1 is the first mammalian member of the betagamma superfamily with a primarily non-lens role. Other parts of the predicted AIM1 protein sequence have weak similarity with filament or actin-binding proteins. AIM1 is a good candidate for the putative suppressor of malignant melanoma on chromosome 6, possibly exerting its effects through interactions with the cytoskeleton.


Cluster Monte Carlo algorithm for the quantum rotor model.
We propose a highly efficient "worm"-like cluster Monte Carlo algorithm for the quantum rotor model in the link-current representation. We explicitly prove detailed balance for the algorithm even in the presence of disorder. For the pure quantum rotor model with mu=0, the algorithm yields high- precision estimates for the critical point K(c)=0.333 05(5) and the correlation length exponent nu=0.670(3). For the disordered case, mu=1 / 2+/-1 / 2, we find nu=1.15(10).


Autofluorescent Granules of the Human Retinal Pigment Epithelium: Phenotypes, Intracellular Distribution, and Age-Related Topography.
The human retinal pigment epithelium (RPE) accumulates granules significant for autofluorescence imaging. Knowledge of intracellular accumulation and distribution is limited. Using high-resolution microscopy techniques, we determined the total number of granules per cell, intracellular distribution, and changes related to retinal topography and age. RPE cells from the fovea, perifovea, and near-periphery of 15 human RPE flat mounts were imaged using structured illumination microscopy (SIM) and confocal fluorescence microscopy in young (≤51 years, n = 8) and older (>80 years, n = 7) donors. Using custom FIJI plugins, granules were marked with computer assistance, classified based on morphological and autofluorescence properties, and analyzed with regard to intracellular distribution, total number per cell, and granule density. A total of 193,096 granules in 450 RPE cell bodies were analyzed. Based on autofluorescence properties, size, and composition, the RPE granules exhibited nine different phenotypes (lipofuscin, two; melanolipofuscin, five; melanosomes, two), distinguishable by SIM. Overall, lipofuscin (low at the fovea but increases with eccentricity and age) and melanolipofuscin (equally distributed at all three locations with no age-related changes) were the major granule types. Melanosomes were under-represented due to suboptimal visualization of apical processes in flat mounts. Low lipofuscin and high melanolipofuscin content within foveal RPE cell bodies and abundant lipofuscin at the perifovea suggest a different genesis, plausibly related to the population of overlying photoreceptors (fovea, cones only; perifovea, highest rod density). This systematic analysis provides further insight into RPE cell and granule physiology and links granule load to cell autofluorescence, providing a subcellular basis for the interpretation of clinical fundus autofluorescence.


Simultaneous application of interstitial flow and cyclic mechanical strain to a three-dimensional cell-seeded hydrogel.
The present study describes the design and validation of a simple apparatus to apply simultaneous mechanical and fluidic stress to three-dimensional (3D) cell-seeded collagen hydrogels. Constructs were formed in wells in a silicone substrate that could be stretched cyclically, and were also fitted with inlet ports to apply fluid flow. Acid etching was used to retain adhesion of the gels to the walls of the well, and an acellular layer of collagen hydrogel was used to distribute flow evenly. Finite element modeling showed that 5% uniaxial strain applied to the entire silicone substrate resulted in ∼6.5% strain in each of the gel constructs. Permeability testing and flow observation showed that acellular hydrogels were fourfold more permeable than cardiac fibroblast-seeded gels, and that the fluid distributed evenly in the acellular layer before entering the cell-seeded gel. Viability testing and imaging demonstrated that cells remained viable with expected fibroblast morphology for the 120 h duration of the experiments. These results demonstrate that this simple bioreactor can be used to study the effects of mechanical strain and interstitial flow in 3D protein hydrogels. Such 3D tissue models have utility in studying cell and tissue responses to their mechanical environment.


Depression, Anxiety, and Patterns of Mental Health Care Among Men With Prostate Cancer Receiving Androgen Deprivation Therapy.
Androgen deprivation therapy (ADT) use is associated with an increased risk of developing depression and anxiety. Little is known about how the mental health of these men is treated. We identified men with prostate cancer who received ADT between 2001 and 2015 using Optum's de-identified Clinformatics Data Mart Database. We determined the incidence of depression or anxiety diagnoses, mental health treatments, and the specialty of providers initiating psychotropic medications, after the start of ADT. Outcomes were compared with those of men with prostate cancer not receiving ADT and men without prostate cancer. Of 37 388 men with prostate cancer treated with ADT, 3964 (10.6%) received a new diagnosis of depression or anxiety. Of those 3964 men, 1892 (47.7%) did not receive a documented treatment, 10 (0.3%) received psychotherapy, 1321 (33.3%) a selective serotonin reuptake inhibitor, and 744 (18.8%) a benzodiazepine. The median time from initiation of ADT to a depression or anxiety diagnosis was 9.3 months. Primary care physicians were the most common prescribers of psychotropic medications (72.2%). The proportion of men not receiving mental health treatments of interest (47.7%) was similar compared to men without prostate cancer (49.1%), but statistically significantly lower compared to men with prostate cancer not receiving ADT (52.7%). In men with prostate cancer receiving ADT with a new diagnosis of depression or anxiety, nearly half are not receiving mental health care while one in five is introduced to a benzodiazepine. Further investigation toward improving the mental health care for men on ADT is needed.


Genome-wide association study in alopecia areata implicates both innate and adaptive immunity.
Alopecia areata (AA) is among the most highly prevalent human autoimmune diseases, leading to disfiguring hair loss due to the collapse of immune privilege of the hair follicle and subsequent autoimmune attack. The genetic basis of AA is largely unknown. We undertook a genome-wide association study (GWAS) in a sample of 1,054 cases and 3,278 controls and identified 139 single nucleotide polymorphisms that are significantly associated with AA (P <or= 5 x 10(-7)). Here we show an association with genomic regions containing several genes controlling the activation and proliferation of regulatory T cells (T(reg) cells), cytotoxic T lymphocyte-associated antigen 4 (CTLA4), interleukin (IL)-2/IL-21, IL-2 receptor A (IL-2RA; CD25) and Eos (also known as Ikaros family zinc finger 4; IKZF4), as well as the human leukocyte antigen (HLA) region. We also find association evidence for regions containing genes expressed in the hair follicle itself (PRDX5 and STX17). A region of strong association resides within the ULBP (cytomegalovirus UL16-binding protein) gene cluster on chromosome 6q25.1, encoding activating ligands of the natural killer cell receptor NKG2D that have not previously been implicated in an autoimmune disease. By probing the role of ULBP3 in disease pathogenesis, we also show that its expression in lesional scalp from patients with AA is markedly upregulated in the hair follicle dermal sheath during active disease. This study provides evidence for the involvement of both innate and acquired immunity in the pathogenesis of AA. We have defined the genetic underpinnings of AA, placing it within the context of shared pathways among autoimmune diseases, and implicating a novel disease mechanism, the upregulation of ULBP ligands, in triggering autoimmunity.


Hepatitis C: rapid progress in medicine and implications for dentistry.
Recent developments in the recognition and diagnosis of the major infectious cause of posttransfusion hepatitis, hepatitis C virus (HCV), have led to an explosion in research. These developments are of relevance to dental providers. Patients may now present to the dentist with a diagnosis of hepatitis C infection and may be undergoing medical treatment for this disease. The risk for transmission of HCV in the dental setting is minimal. The dentist must understand the implications for the diagnosis of HCV for the patient and for the provision of dental care.


Girls in the boat: Sex differences in rowing performance and participation.
Men outperform women in many athletic endeavors due to physiological and anatomical differences (e.g. larger and faster muscle); however, the observed sex differences in elite athletic performance are typically larger than expected, and may reflect sex-related differences in opportunity or incentives. As collegiate rowing in the United States has been largely incentivized for women over the last 20 years, but not men, the purpose of this study was to examine sex differences in elite rowing performance over that timeframe. Finishing times from grand finale races for collegiate championship on-water performances (n = 480) and junior indoor performances (n = 1,280) were compared between men and women across 20 years (1997-2016), weight classes (heavy vs. lightweight) and finishing place. Participation of the numbers of men and women rowers were also quantified across years. Men were faster than women across all finishing places, weight classes and years of competition and performance declined across finishing place for both men and women (P<0.001). Interestingly, the reduction in performance time across finishing place was greater (P<0.001) for collegiate men compared to women in the heavyweight division. This result is opposite to other sports (e.g. running and swimming), and to lightweight rowing in this study, which provides women fewer incentives than in heavyweight rowing. Correspondingly, participation in collegiate rowing has increased by ~113 women per year (P<0.001), with no change (P = 0.899) for collegiate men. These results indicate that increased participation and incentives within collegiate rowing for women vs. men contribute to sex differences in athletic performance.


Effect of an enhanced medical home on serious illness and cost of care among high-risk children with chronic illness: a randomized clinical trial.
Patient-centered medical homes have not been shown to reduce adverse outcomes or costs in adults or children with chronic illness. To assess whether an enhanced medical home providing comprehensive care prevents serious illness (death, intensive care unit [ICU] admission, or hospital stay >7 days) and/or reduces costs among children with chronic illness. Randomized clinical trial of high-risk children with chronic illness (≥3 emergency department visits, ≥2 hospitalizations, or ≥1 pediatric ICU admissions during previous year, and >50% estimated risk for hospitalization) treated at a high-risk clinic at the University of Texas, Houston, and randomized to comprehensive care (n = 105) or usual care (n = 96). Enrollment was between March 2011 and February 2013 (when predefined stopping rules for benefit were met) and outcome evaluations continued through August 31, 2013. Comprehensive care included treatment from primary care clinicians and specialists in the same clinic with multiple features to promote prompt effective care. Usual care was provided locally in private offices or faculty-supervised clinics without modification. Primary outcome: children with a serious illness (death, ICU admission, or hospital stay >7 days), costs (health system perspective). Secondary outcomes: individual serious illnesses, medical services, Medicaid payments, and medical school revenues and costs. In an intent-to-treat analysis, comprehensive care decreased both the rate of children with a serious illness (10 per 100 child-years vs 22 for usual care; rate ratio [RR], 0.45 [95% CI, 0.28-0.73]), and total hospital and clinic costs ($16,523 vs $26,781 per child-year, respectively; cost ratio, 0.58 [95% CI, 0.38-0.88]). In analyses of net monetary benefit, the probability that comprehensive care was cost neutral or cost saving was 97%. Comprehensive care reduced (per 100 child-years) serious illnesses (16 vs 44 for usual care; RR, 0.33 [95% CI, 0.17-0.66]), emergency department visits (90 vs 190; RR, 0.48 [95% CI, 0.34-0.67]), hospitalizations (69 vs 131; RR, 0.51 [95% CI, 0.33-0.77]), pediatric ICU admissions (9 vs 26; RR, 0.35 [95% CI, 0.18-0.70]), and number of days in a hospital (276 vs 635; RR, 0.36 [95% CI, 0.19-0.67]). Medicaid payments were reduced by $6243 (95% CI, $1302-$11,678) per child-year. Medical school losses (costs minus revenues) increased by $6018 (95% CI, $5506-$6629) per child-year. Among high-risk children with chronic illness, an enhanced medical home that provided comprehensive care to promote prompt effective care vs usual care reduced serious illnesses and costs. These findings from a single site of selected patients with a limited number of clinicians require study in larger, broader populations before conclusions about generalizability to other settings can be reached. clinicaltrials.gov Identifier: NCT02128776.


Quorum-sensing molecule dihydroxy-2,3-pentanedione and its analogs as regulators of epithelial integrity.
Quorum-sensing molecules regulate the behavior of bacteria within biofilms and at the same time elicit an immune response in host tissues. Our aim was to investigate the regulatory role of dihydroxy-2,3-pentanedione (DPD), the precursor of universal autoinducer-2 (AI-2), and its analogs (ethyl-DPD, butyl-DPD and isobutyl-DPD) in the integrity of gingival epithelial cells. Human gingival keratinocytes were incubated with four concentrations (10 μmol L-1 , 1 μmol L-1 , 100 nmol L-1 and 10 nmol L-1 ) of DPD and its analogs for 24 hours. The numbers of viable cells were determined using a proliferation kit, matrix metalloproteinase (MMP)-2 and -9 activities were determined by gelatin zymography, and expression of occludin protein and occludin mRNA were determined by western blotting and RT-qPCR, respectively. Increased cell proliferation was observed in gingival keratinocytes incubated with 100 nmol L-1 of butyl-DPD. MMP-9 activity was elevated in cells incubated with 10 μmol L-1 of ethyl-DPD. On the other hand, MMP-2 activity did not show any significant change when gingival keratinocytes were incubated with or without DPD or analogs. Western blot analyses demonstrated five forms (105, 61, 52.2, 44 and 37 kDa) of occludin. Incubation with 1 μmol L-1 and 100 nmol L-1 of DPD and with 10 nmol L-1 of ethyl-DPD increased dimeric (105 kDa) forms of occludin, while incubation with 100 nmol L-1 of isobutyl-DPD increased monomeric (61 kDa) forms. DPD and ethyl-DPD decreased, and 100 nmol L-1 of isobutyl-DPD and 10 nmol L-1 of butyl-DPD increased, the monomeric (52.2 kDa and 44 kDa) forms of occludin, whereas ethyl-DPD decreased and isobutyl-DPD increased, the low-molecular-weight (37 kDa) forms. According to RT-qPCR analysis, the exposure of gingival keratinocytes to 10 μmol L-1 of isobutyl-DPD up-regulated expression of occludin. The results indicate that isobutyl-DPD has the potential to enhance the integrity of the epithelium by stimulating the formation of occluding, without affecting the proliferation or gelatinolytic enzyme activities of the exposed cells. The modulatory effect of an AI-2 analog on the epithelial cell response is shown for the first time.


Optic nerve size in traumatic optic neuropathy.
The assessment of traumatic optic neuropathy in a neurologically impaired patient is difficult, and the size of the optic nerve has been suggested as a potential adjunct in differential diagnosis. This controlled retrospective study was designed to evaluate the size of the optic nerve involved in traumatic optic neuropathy when compared to the optic nerve in the noninjured eye. Maxillofacial computerized tomographs of 22 patients were examined; intraorbital optic nerve diameter was measured on injured and noninjured sides, with the examiner unaware of the side of injury. No statistically significant difference in size was found between the traumatized optic nerves and those that remained intact. The authors have concluded that the size of the optic nerve is not a reliable predicator of the presence or absence of traumatic optic neuropathy when measured with computerized tomograph scans.


Expression of microRNAs and their target mRNAs in human stem cell-derived cardiomyocyte clusters and in heart tissue.
Recent studies have shown that microRNAs (miRNAs) act as posttranscriptional regulators and that they play important roles during heart development and in cardiac function. Thus, they may provide new means of altering stem cell fate and differentiation processes. However, information about the correlation between global miRNA and mRNA expression in cardiomyocyte clusters (CMCs) derived from human embryonic stem cells (hESC) and in fetal and adult heart tissue is lacking. In the present study the global miRNA and mRNA expression in hESC-derived CMCs and in fetal and adult heart tissue was investigated in parallel using microarrays. Target genes for the differentially expressed miRNAs were predicted using computational methods, and the concordance in miRNA expression and mRNA levels of potential target genes was determined across the experimental samples. The biology of the predicted target genes was further explored regarding their molecular functions and involvement in known regulatory pathways. A clear correlation between the global miRNA expression and corresponding target mRNA expression was observed. Using three different sources of cardiac tissue-like samples, we defined the similarities between in vitro hESC-derived CMCs and their in vivo counterparts. The results are in line with previously reported observations that miRNAs repress mRNA expression and additionally identify a number of novel miRNAs with potential important roles in human cardiac tissue. The concordant miRNA expression pattern observed among all the cardiac tissue-like samples analyzed here provide a starting point for future ambitious studies aiming towards assessment of the functional roles of specific miRNAs during cardiomyocyte differentiation.


[Persistent udder edema as a herd problem: a literature review].
The data presented in literature suggest that disturbances of volume regulation do not have a role in the pathogenesis of persistent udder oedema. Instead, they favour local mammary problems. The presence of both mast cells and oedema, especially in subcutaneous tissue, focuses attention on the role of mast cells in the pathogenesis of persistent udder oedema via increased endothelial permeability due to histamine release. The dissociation constants (Kd) for the high-affinity glucocorticoid receptor of normal and oedematous udders are not significantly different, and nor are there differences between the total quantity of dexamethasone bound (Bmax) and mast cell density in normal and oedematous udders. It has been shown that, in cattle, the concentration of histamine is higher in milk than in plasma. In addition, the urine histamine concentration is affected by the diet. Treatments to reduce udder oedema include combined salt and water restriction, supplementation with potassium or vitamin E (alpha-tocopheryl acetate), sodium restriction, and administration of hydrochlorothiazide intramuscularly, chlorothiazide orally, or sodium-acetazolamide orally and/or parenterally (i.v./i.m.). A low cation-anion balance in a diet containing 1.6% calcium also seems to reduce mammary oedema formation.


Are we able to correctly identify prostate cancer patients who could be adequately treated by focal therapy?
Because of the improvements on detection of early stage prostate cancer over the last decade, focal therapy for localized prostate cancer (PC) has been proposed for patients with low-risk disease. Such treatment would allow the control of cancer, thereby diminishing side effects, such as urinary incontinence and sexual dysfunction, which have an enormous impact on quality of life. The critical issue is whether it is possible to preoperatively predict clinically significant unifocal or unilateral prostate cancer with sufficient accuracy. Our aim is to determine whether there is any preoperative feature that can help select the ideal patient for focal therapy. A total of 599 patients who underwent transrectal ultrasound (TRUS)-guided prostate biopsy followed by radical prostatectomy to treat PC were examined in our laboratory between 2001 and 2009. We established very restricted criteria to select patients with very-low-risk disease for whom focal therapy would be suitable (only 1 biopsy core positive, tumor no larger than 80% of a single core, no perineural invasion, PSA serum level < 10 ng/ml, Gleason score < 7 and clinical stage T1c, T2a-b). We defined 2 groups of patients who would be either adequately treated or not treated by focal therapy. The primary endpoint was the evaluation of preoperative features in order to identify which parameters should be considered when choosing good candidates for focal therapy. Fifty-six out of 599 patients met our criteria. The mean age was 59 years, and the mean number of biopsy cores was 14.4. Forty-seven (83.9%) were staged T1c, and 9 (16.1%) were staged T2a-b. Forty-four (78.6%) patients could be considered to have been adequately treated by focal therapy, and 12 (21.4%) could not. There was no statistical difference between the 2 groups considering age, clinical stage, PSA levels, Gleason score, and tumor volume in the biopsy. All 12 patients who could be considered inadequately treated had a bilateral, significant secondary tumor, 58.3% had Gleason ≥ 7, and 25% were staged pT3. Although focal therapy might be a good option for patients with localized prostate cancer, we are so far unable to select which of them would benefit from it based on preoperative data, even using very restricted criteria, and a considerable proportion of men would still be left undertreated.


Tau seeding and spreading in vivo is supported by both AD-derived fibrillar and oligomeric tau.
Insoluble fibrillar tau, the primary constituent of neurofibrillary tangles, has traditionally been thought to be the biologically active, toxic form of tau mediating neurodegeneration in Alzheimer's disease. More recent studies have implicated soluble oligomeric tau species, referred to as high molecular weight (HMW) due to its properties on size exclusion chromatography, in tau propagation across neural systems. These two forms of tau have never been directly compared. We prepared sarkosyl insoluble and HMW tau from the frontal cortex of Alzheimer patients and compared their properties using a variety of biophysical and bioactivity assays. Sarkosyl insoluble fibrillar tau is comprised of abundant paired helical filaments (PHF) as quantified by electron microscopy (EM), and is more resistant to proteinase K, compared to HMW tau which is mostly in an oligomeric form. Sarkosyl insoluble and HMW tau are nearly equivalent in potency in a HEK cell bioactivity assay for seeding aggregates and their injection reveals similar local uptake into hippocampal neurons in PS19 Tau transgenic mice. However, the HMW preparation appears to be far more potent in inducing a glial response including Clec7a-positive rod-microglia in the absence of neurodegeneration or synapse loss and promotes more rapid propagation of misfolded tau to distal, anatomically connected regions, such as entorhinal and perirhinal cortices. These data suggest that soluble HMW tau has similar properties to fibrillar sarkosyl insoluble tau with regard to tau seeding potential but may be equal or even more bioactive with respect to propagation across neural systems and activation of glial responses, both relevant tau-related Alzheimer phenotypes.


Expression and purification of functional recombinant epitopes for the platelet antigens, PlA1 and PlA2.
The platelet antigens, PlA1 and PlA2, are responsible for most cases of posttransfusion purpura (PTP) and neonatal alloimmune thrombocytopenia (NAIT) in the caucasian population and are determined by two allelic forms of the platelet glycoprotein GPIIIa gene. To study the interaction between these antigens and their respective antibodies, we inserted the sequence that encodes the signal peptide and the N-terminal 66 amino acids of the PlA1 form of GPIIIa into the expression vector pGEX1. To express the PlA2 antigen, nucleotide 196 of the PlA1 coding sequence was mutated to the PlA2 allelic form. When transformed and induced in Escherichia coli, the two constructs produce glutathione S-transferase (GST)/N-terminal GPIIIa fusion proteins, one containing leucine at position 33 (PlA1), the other proline (PlA2). These proteins are easily purified in milligram quantities using glutathione-Sepharose and react specifically with their respective antibodies by immunoblot and enzyme-linked immunosorbent assay. Antigenicity of the PlA1 fusion protein in reduced glutathione increases with time; moreover, the addition of oxidized glutathione accelerates this process, presumably because of formation of the native disulfide bonds. Neutralization assays indicate that the PlA1 fusion protein competes for all of the anti-PlA1 antibody in the serum of patients with PTP and NAIT that is capable of interacting with the surface of intact platelets. This study shows that the GST/N-terminal GPIIIa fusion proteins contain conformational epitopes that mimic those involved in alloimmunization, and that regions other than the amino terminal 66 amino acids of GPIIIa are not likely to contain or be required for the development of functional PlA1 epitopes. Furthermore, these recombinant proteins can be used for the affinity-purification of clinical anti-PlA1 antibodies and specific antibody identification by western blotting, making them useful in the diagnosis of patients alloimmunized to PlA1 alloantigens.


BiG-MAP: an Automated Pipeline To Profile Metabolic Gene Cluster Abundance and Expression in Microbiomes.
Microbial gene clusters encoding the biosynthesis of primary and secondary metabolites play key roles in shaping microbial ecosystems and driving microbiome-associated phenotypes. Although effective approaches exist to evaluate the metabolic potential of such bacteria through identification of these metabolic gene clusters in their genomes, no automated pipelines exist to profile the abundance and expression levels of such gene clusters in microbiome samples to generate hypotheses about their functional roles, and to find associations with phenotypes of interest. Here, we describe BiG-MAP, a bioinformatic tool to profile abundance and expression levels of gene clusters across metagenomic and metatranscriptomic data and evaluate their differential abundance and expression under different conditions. To illustrate its usefulness, we analyzed 96 metagenomic samples from healthy and caries-associated human oral microbiome samples and identified 252 gene clusters, including unreported ones, that were significantly more abundant in either phenotype. Among them, we found the muc operon, a gene cluster known to be associated with tooth decay. Additionally, we found a putative reuterin biosynthetic gene cluster from a Streptococcus strain to be enriched but not exclusively found in healthy samples; metabolomic data from the same samples showed masses with fragmentation patterns consistent with (poly)acrolein, which is known to spontaneously form from the products of the reuterin pathway and has been previously shown to inhibit pathogenic Streptococcus mutans strains. Thus, we show how BiG-MAP can be used to generate new hypotheses on potential drivers of microbiome-associated phenotypes and prioritize the experimental characterization of relevant gene clusters that may mediate them. IMPORTANCE Microbes play an increasingly recognized role in determining host-associated phenotypes by producing small molecules that interact with other microorganisms or host cells. The production of these molecules is often encoded in syntenic genomic regions, also known as gene clusters. With the increasing numbers of (multi)omics data sets that can help in understanding complex ecosystems at a much deeper level, there is a need to create tools that can automate the process of analyzing these gene clusters across omics data sets. This report presents a new software tool called BiG-MAP, which allows assessing gene cluster abundance and expression in microbiome samples using metagenomic and metatranscriptomic data. Here, we describe the tool and its functionalities, as well as its validation using a mock community. Finally, using an oral microbiome data set, we show how it can be used to generate hypotheses regarding the functional roles of gene clusters in mediating host phenotypes.


Knowledge, attitudes, and practices regarding rabies in Grenada.
While Grenada attained a zero-human-rabies case status since 1970, the authors conducted the first study to assess knowledge, attitudes, and practices that may contribute to this status as well as to receive feedback on the rabies control program in Grenada. A cross-sectional survey was conducted in July, 2017 with 996 households on the mainland. A questionnaire was administered to collect information on knowledge of rabies and prevention, vaccination practices, perception of institutional responsibilities for rabies control, and evaluation of the anti-rabies program. Of the 996 households, 617 (62%) had owners of animals that can be infected with rabies and were included in the analysis. Respondents were very aware of rabies as a disease that can infect animals and humans. The rate of participation in the vaccination program was 51.6% for pets and 38.0% for livestock. About 40% of respondents were knowledgeable about the extent of protection from the rabies vaccine. Respondents did not demonstrate exceptionally high levels of knowledge about animals that were likely to be infected with rabies, neither the anti-rabies programs that were conducted in Grenada. The three most frequent recommendations made to improve the rabies-control programs were: increase education programs, control the mongoose population, and expand the vaccination period each year. Conducting a comprehensive national rabies education program, expanding the vaccination program, and increasing the rate of animal vaccination are important steps that need to be taken to maintain the current zero-human-case status.


'They would never receive you without a husband': Paradoxical barriers to antenatal care scale-up in Rwanda.
To explore perspectives and experiences of antenatal care and partner involvement among women who nearly died during pregnancy ('near-miss'). A study guided by naturalistic inquiry was conducted, and included extended in-community participant observation, semi-structured interviews, and focus group discussions. Qualitative data were collected between March 2013 and April 2014 in Kigali, Rwanda. All informants were aware of the recommendations of male involvement for HIV-testing at the first antenatal care visit. However, this recommendation was seen as a clear link in the chain of delays and led to severe consequences, especially for women without engaged partners. The overall quality of antenatal services was experienced as suboptimal, potentially missing the opportunity to provide preventive measures and essential health education intended for both parents. This seemed to contribute to women's disincentive to complete all four recommended visits and men's interest in attending to ensure their partners' reception of care. However, the participants experienced a restriction of men's access during subsequent antenatal visits, which made men feel denied to their increased involvement during pregnancy. 'Near-miss' women and their partners face paradoxical barriers to actualise the recommended antenatal care visits. The well-intended initiative of male partner involvement counterproductively causes delays or excludes women whereas supportive men are turned away from further health consultations. Currently, the suboptimal quality of antenatal care misses the opportunity to provide health education for the expectant couple or to identify and address early signs of complications These findings suggest a need for increased flexibility in the antenatal care recommendations to encourage women to attend care with or without their partner, and to create open health communication about women's and men's real needs within the context of their social situations. Supportive partners should not be denied involvement at any stage of pregnancy, but should be received only upon consent of the expectant mother.


Expression of survivin does not appear to influence breast cancer recurrence risk.
Survivin is an inhibitor of apoptosis, and its expression associates with poor outcomes in multiple cancers. It may be a therapeutic target due to its unique expression in cancer cells. We estimated the association between nuclear and cytoplasmic survivin expression in primary tumors and breast cancer recurrence. In this case-control study, we included women age 35-69, diagnosed with stage I-III breast cancer between 1985 and 2001, and registered with the Danish Breast Cancer Group. We identified 541 patients with breast cancer recurrence with estrogen receptor-positive disease who were treated with tamoxifen for at least 1 year (ER+/TAM+) and 300 with estrogen receptor-negative carcinomas, not treated with tamoxifen, and who survived at least 1 year (ER-/TAM-). Controls were matched to cases on ER/TAM status, date of surgery, menopausal status, stage and county. Survivin expression was estimated by immunohistochemistry on tissue microarrays. We fit logistic regression models to estimate odds ratios (ORs) and 95% confidence intervals (CIs) associating nuclear and cytoplasmic survivin expression with recurrence. Associations between nuclear and cytoplasmic survivin expression and breast cancer recurrence were near-null in both ER+/TAM + and ER-/TAM - strata. For the cytoplasmic to nuclear ratio (CNR) of survivin expression, we found a null association in the ER+/TAM + group comparing CNR ≥5 with CNR <5, but an association (OR =2.48, 95% CI: 1.15, 5.31) in the ER-/TAM - group. Survivin expression was not associated with breast cancer recurrence in this study. The CNR ratio may warrant further investigation especially among ER - tumors.


Von Hippel-Lindau tumor suppressor protein transforms human neuroblastoma cells into functional neuron-like cells.
Von Hippel-Lindau (VHL) tumor suppressor protein is expressed in neurons of the central nervous system and plays an important role during the neuronal differentiation of central nervous system progenitor cells. To elucidate the neuronal differentiating potential of VHL protein in neuroblastoma cells, we overexpressed or inhibited VHL protein in human neuroblastoma cells (SY-SH5Y), and examined the morphological change, expressions of neuronal markers, and electrophysiological functions. Here we show that with VHL gene transduction SY-SH5Y cells stably expressing the VHL protein had neurite-like processes with varicosities, showed the distinct expression of the neuronal markers neuropeptide Y and neurofilament 200, acquired regulated neurosecretion competence in response to depolarizing and cholinergic stimuli, and had large voltage-gated fast sodium currents and delayed rectifier potassium (Kv) currents compatible with those of functional neurons. In addition, they displayed inactivated ether-á-go-go potassium channels related to the promotion of the cell cycle and to the termination of differentiation. Also, by treatment with retinoic acid, they rapidly underwent cell death related to apoptosis. These findings suggest that the induction of neuronal function by VHL protein is associated with down-regulation of the cell cycle. In contrast, the inhibition of endogenous expression of VHL protein with antisense-orientated VHL gene transduction reduced such neuronal properties inherent to these cells, including the capacity for activation of ether-á-go-go channels. In conclusion, VHL protein has a neuronal differentiating potential to transform neuroblastoma cells into functional neuron-like cells. Our finding of the neuronal differentiation of neuroblastoma cells under the control of the VHL gene may contribute to the development of clinical techniques for neuronal regeneration in the case of intractable neuronal diseases and for differentiation therapy against neuroblastomas.


The prevalence and correlates of suicidal ideation in Chinese psychiatric patients during the fifth wave of COVID-19 in Hong Kong.
People with mental disorders have increased risk of psychological distress during COVID-19. However, there is limited research comprehensively examining factors associated with suicidal ideation, the strongest predictor of suicidal behavior, among psychiatric patients amidst pandemic. We investigated prevalence and correlates of suicidal ideation in 407 Chinese psychiatric outpatients (diagnosed with mood, anxiety or schizophrenia-spectrum disorders) aged 18-64 years during the peak of fifth COVID-19 wave in Hong Kong between 28 March and 8 April, 2022, based on a comprehensive array of variables encompassing socio-demographics, illness profile, psychopathological symptoms, psychological measures and pandemic-related factors. Univariate and multivariate logistic regression analyses were conducted to determine correlates of suicidal ideation. Results showed that 128 (31.4%) participants exhibited suicidal ideation. Univariate analyses revealed that being unemployed or full-time student, more severe depressive, anxiety, PTSD-like, insomnia and psychotic symptoms, higher levels of loneliness, avoidant-coping, greater pandemic-related stress burden and distress by social-distancing measures were related to suicidal ideation. Conversely, participants with higher monthly household-income, better quality-of-life, and greater resilience were less likely to have suicidal ideation. Notably, only depressive symptom severity was retained in final multivariate model as a factor significantly associated with suicidal ideation. Hence, we observed that approximately one-third of Chinese psychiatric patients experienced suicidal ideation during fifth pandemic wave. Our findings underscore the influence of depressive symptoms being above and beyond that of other psychopathological symptoms, psychological and pandemic-related variables on suicidal ideation. Longitudinal research is required to clarify suicidal ideation trajectories and predictors of persistent suicidal ideation across pandemic and post-pandemic periods.


The highly virulent 2006 Norwegian EHEC O103:H25 outbreak strain is related to the 2011 German O104:H4 outbreak strain.
In 2006, a severe foodborne EHEC outbreak occured in Norway. Seventeen cases were recorded and the HUS frequency was 60%. The causative strain, Esherichia coli O103:H25, is considered to be particularly virulent. Sequencing of the outbreak strain revealed resemblance to the 2011 German outbreak strain E. coli O104:H4, both in genome and Shiga toxin 2-encoding (Stx2) phage sequence. The nucleotide identity between the Stx2 phages from the Norwegian and German outbreak strains was 90%. During the 2006 outbreak, stx(2)-positive O103:H25 E. coli was isolated from two patients. All the other outbreak associated isolates, including all food isolates, were stx-negative, and carried a different phage replacing the Stx2 phage. This phage was of similar size to the Stx2 phage, but had a distinctive early phage region and no stx gene. The sequence of the early region of this phage was not retrieved from the bacterial host genome, and the origin of the phage is unknown. The contaminated food most likely contained a mixture of E. coli O103:H25 cells with either one of the phages.


Biomimetic Virulomics for Capture and Identification of Cell-Type Specific Effector Proteins.
An unmet challenge in the study of disease is to accurately streamline the identification of important virulence factors. Traditional, genetically driven approaches miss biologically relevant markers due to discordance between the genome and proteome. Here, we developed a nanotechnology-enabled affinity enrichment strategy coupled with multiplexed quantitative proteomics, namely Biomimetic Virulomics, for successful identification of cell-type specific effector proteins of both prokaryotic and eukaryotic pathogens. We highlight the power of Biomimetic Virulomics by capturing known virulence factors in a high-throughput, cell-type guided fashion. Additionally, a comprehensive characterization of the membrane protein component of biomimetics utilized in this strategy is provided. Interfacing cell-derived nanomaterials with multiplexed quantitative proteomics allow for a specific targeting strategy of virulence factors that can be utilized for drug discovery against prominent human diseases.


Developing the content of a locomotor disability scale for adults in Bangladesh: a qualitative study.
Bangladesh has an estimated 17 million adults with disabilities. A significant proportion of them are believed to have locomotor disabilities. There are over 300 non-governmental organizations providing different types of rehabilitation services to them. However, there is no locally developed and validated locomotor disability measurement scale in Bangladesh. The purpose of this study was to develop a locomotor disability scale with disability indicators suitable for adults in Bangladesh. Semi-structured interviews were conducted with 25 purposively selected adults with locomotor disabilities to generate scale items. At the second stage, cognitive interviews were conducted with 12 purposively selected adults with locomotor disabilities in order to refine the measurement questions and response categories. Data were analysed using the framework technique- identifying, abstracting, charting and matching themes across the interviews. For a locomotor disability scale, 70 activities (disability indicators) were identified: 37 mobility activities, 9 activities of daily living, 17 work/productivity activities and 7 leisure activities. Cognitive interviews revealed that when asking the respondents to rate their difficulty in performing the activities, instead of just mentioning the activity name, such as taking a bath or shower, a detailed description of the activity and response options were necessary to ensure consistent interpretation of the disability indicators and response options across all respondents. Identifying suitable disability indicators was the first step in developing a locomotor disability scale for adults in Bangladesh. Interviewing adults with locomotor disabilities in Bangladesh ensured that the locomotor disability scale is of relevance to them and consequently it has excellent content validity. Further research is needed to evaluate the psychometric properties of this scale.


Evidence of AlOHC responsible for the radiation-induced darkening in Yb doped fiber.
Using a combination of experimental techniques such as optical absorption, Raman scattering, continuous wave and pulse Electron Spin Resonance (ESR), we characterize a set of γ-irradiated Yb(3+) doped silica glass preforms with different contents of phosphorous and aluminum. We demonstrate that when P is introduced in excess compared to Al, nearly no radiodarkening is induced by γ-rays. On the other hand, when Al>P, a large absorption band is induced by radiation. Thermal annealing experiments reveal the correlation between the decrease of the optical absorption band and the decrease of the Al-Oxygen Hole Center (AlOHC) ESR signal, demonstrating the main role of AlOHC defects in the fiber darkening. HYSCORE (HYperfine Sublevel CORElation) pulse-ESR experiments show a high Al-P nuclear spin coupling when P>Al and no coupling when Al>P. This result suggests that both AlOHC and POHC creation is inhibited by Al-O-P linkages. Confronting our data with previous works, we show that the well-known photodarkening process, meaning losses induced by the IR pump, can also be explained in this framework.


Two novel types of O-glycans on the mugwort pollen allergen Art v 1 and their role in antibody binding.
Art v 1, the major allergen of mugwort (Artemisia vulgaris) pollen contains galactose and arabinose. As the sera of some allergic patients react with natural but not with recombinant Art v 1 produced in bacteria, the glycosylation of Art v 1 may play a role in IgE binding and human allergic reactions. Chemical and enzymatic degradation, mass spectrometry, and 800 MHz (1)H and (13)C nuclear magnetic resonance spectroscopy indicated the proline-rich domain to be glycosylated in two ways. We found a large hydroxyproline-linked arabinogalactan composed of a short beta1,6-galactan core, which is substituted by a variable number (5-28) of alpha-arabinofuranose residues, which form branched side chains with 5-, 2,5-, 3,5-, and 2,3,5-substituted arabinoses. Thus, the design of the Art v 1 polysaccharide differs from that of the well known type II arabinogalactans, and we suggest it be named type III arabinogalactan. The other type of glycosylation was formed by single (but adjacent) beta-arabinofuranoses linked to hydroxyproline. In contrast to the arabinosylation of Ser-Hyp(4) motifs in other hydroxyproline-rich glycoproteins, such as extensins or solanaceous lectins, no oligo-arabinosides were found in Art v 1. Art v 1 and parts thereof produced by alkaline degradation, chemical deglycosylation, proteolytic degradation, and/or digestion with alpha-arabinofuranosidase were used in enzyme-linked immunosorbent assay and immunoblot experiments with rabbit serum and with the sera of patients. Although we could not observe antibody binding by the polysaccharide, the single hydroxyproline-linked beta-arabinose residues appeared to react with the antibodies. Mono-beta-arabinosylated hydroxyproline residues thus constitute a new, potentially cross-reactive, carbohydrate determinant in plant proteins.


Use of Bacteroidales microbial source tracking to monitor fecal contamination in fresh produce production.
In recent decades, fresh and minimally processed produce items have been associated with an increasing proportion of food-borne illnesses. Most pathogens associated with fresh produce are enteric (fecal) in origin, and contamination can occur anywhere along the farm-to-fork chain. Microbial source tracking (MST) is a tool developed in the environmental microbiology field to identify and quantify the dominant source(s) of fecal contamination. This study investigated the utility of an MST method based on Bacteroidales 16S rRNA gene sequences as a means of identifying potential fecal contamination, and its source, in the fresh produce production environment. The method was applied to rinses of fresh produce, source and irrigation waters, and harvester hand rinses collected over the course of 1 year from nine farms (growing tomatoes, jalapeño peppers, and cantaloupe) in Northern Mexico. Of 174 samples, 39% were positive for a universal Bacteroidales marker (AllBac), including 66% of samples from cantaloupe farms (3.6 log10 genome equivalence copies [GEC]/100 ml), 31% of samples from tomato farms (1.7 log10 GEC/100 ml), and 18% of samples from jalapeño farms (1.5 log10 GEC/100 ml). Of 68 AllBac-positive samples, 46% were positive for one of three human-specific markers, and none were positive for a bovine-specific marker. There was no statistically significant correlation between Bacteroidales and generic Escherichia coli across all samples. This study provides evidence that Bacteroidales markers may serve as alternative indicators for fecal contamination in fresh produce production, allowing for determination of both general contamination and that derived from the human host.


Mycobacteria and human heat shock protein-specific cytotoxic T lymphocytes in rheumatoid synovial inflammation.
To study the cytotoxic capacity of mycobacteria-specific T lymphocyte lines and clones from sites of inflammation in patients with rheumatoid arthritis (RA). We also studied antigen specificity, surface phenotype, expression of T cell receptors (TCR), and HLA restriction. Autologous macrophages (M phi) from the synovial membrane (SM), synovial fluid (SF), or peripheral blood (PB) were used as target cells in cytotoxicity assays. All SM and SF cell lines tested thus far have shown specific lysis of the autologous M phi from SM or PB that had been pulsed with BCG (bacillus Calmette-Guerin), but no cytotoxicity when the targets were pulsed with irrelevant antigens such as tetanus toxoid and Chlamydia. Both CD4+ and CD8+ cells were shown to be involved in the specific cytolysis. The majority of the cytotoxic T lymphocyte (CTL) lines were TCR alpha/beta + cells. However, both TCR alpha/beta + and TCR gamma/delta + clones (TCR delta 1+) from one RA patient showed antigen-specific lysis. Antigen-specific recognition by a number of CTL lines and clones generated from SF and SM was restricted by HLA-DR molecules. Two Mycobacterium bovis 65-kd heat shock protein (HSP)-specific TCR alpha/beta + SF T cell clones also lysed M phi that had been pulsed with a recombinant human 65-kd HSP. Joint inflammation and destruction might be partly attributable to a cross-reaction of mycobacteria-induced cytotoxic T cells with self HSP.


Three-year clinical evaluation of a resin modified glass-ionomer cement and a composite resin in non-carious class V lesions.
The purpose of this study was a 3-year clinical evaluation of a resin modified glass-ionomer and a composite resin restorative material in non-carious class V lesions. In 24 patients 98 non-carious class V lesions were restored with either a resin modified glass-ionomer (Vitremer), or a composite resin restoration (Z100). The restorations were clinically evaluated after 6, 12, 24 and 36 months with the US Public Health Service criteria. At 3 years, 88 teeth of 21 patients were evaluated. All restorations were rated clinically acceptable for colour match, marginal discoloration, marginal adaptation and anatomical form. Restoration retention of both groups was high without any statistically significant difference. However, Vitremer restorations showed a lower incidence of Alfa scores for colour match and marginal discoloration than Z100 restorations (P < 0.05).


Genetic abrogation of immune checkpoints in antigen-specific cytotoxic T-lymphocyte as a potential alternative to blockade immunotherapy.
T cell function can be compromised during chronic infections or through continuous exposure to tumor antigens by the action of immune checkpoint receptors, such as programmed cell death protein 1 (PD-1). Systemic administration of blocking antibodies against the PD-1 pathway can restore T cell function, and has been approved for the treatment of several malignancies, although there is a risk of adverse immune-related side-effects. We have developed a method for generating gene knockouts in human antigen (Ag)-specific cytotoxic T-Lymphocyte (CTLs) using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome editing. Using this method, we generated several transduced CD4+ or CD8+ antigen-specific polyclonal CTL lines and clones, and validated gene modifications of the PD-1 gene. We compared these T-cell lines and clones with control groups in the presence of programmed death-ligand 1 (PD-L1) and observed improved effector functions in the PD1-disrupted cell group. Overall, we have developed a versatile tool for functional genomics in human antigen-specific CTL studies. Furthermore, we provide an alternative strategy for current cell-based immunotherapy that will minimize the side effects caused by antibody blockade therapy.


Ultrathin two-dimensional superconductivity with strong spin-orbit coupling.
We report on a study of epitaxially grown ultrathin Pb films that are only a few atoms thick and have parallel critical magnetic fields much higher than the expected limit set by the interaction of electron spins with a magnetic field, that is, the Clogston-Chandrasekhar limit. The epitaxial thin films are classified as dirty-limit superconductors because their mean-free paths, which are limited by surface scattering, are smaller than their superconducting coherence lengths. The uniformity of superconductivity in these thin films is established by comparing scanning tunneling spectroscopy, scanning superconducting quantum interference device (SQUID) magnetometry, double-coil mutual inductance, and magneto-transport, data that provide average superfluid rigidity on length scales covering the range from microscopic to macroscopic. We argue that the survival of superconductivity at Zeeman energies much larger than the superconducting gap can be understood only as the consequence of strong spin-orbit coupling that, together with substrate-induced inversion-symmetry breaking, produces spin splitting in the normal-state energy bands that is much larger than the superconductor's energy gap.


Gamification: a Novel Approach to Mental Health Promotion.
Gamification has emerged as a novel technique for improving mental health and enhancing treatment effectiveness. This paper provides an overview of gamification approaches to mental health intervention, identifies factors that may be related to variations in treatment effectiveness, and discusses possible strategies for tailoring gamified interventions to clients' needs. Recent research has documented the potential of gamified mental health interventions for bolstering mental wellness and mitigating psychological symptoms. However, their effectiveness may vary depending on study design-related factors and gender-specific considerations. Literature reviews have also identified yet-to-be resolved issues surrounding the possible strengths and weaknesses of the personalization versus standardization of gamification, as well as the potential benefits of gamification for increasing engagement versus the potential risks of over-engagement and behavioral addiction to gamified components. This review highlights the need for careful planning and execution of gamified mental health interventions to optimize their effectiveness and suitability for meeting clients' individual needs and preferences.


Stable isotopes may provide evidence for starvation in reptiles.
Previous studies have attempted to correlate stable isotope signatures of tissues with the nutritional condition of birds, mammals, fishes, and invertebrates. Unfortunately, very little is known about the relationship between food limitation and the isotopic composition of reptiles. We examined the effects that starvation has on delta13C and delta15N signatures in the tissues (excreta, carcass, scales, and claws) of six, distantly related squamate reptiles (gaboon vipers, Bitis gabonica; ball pythons, Python regius; ratsnakes, Elaphe obsoleta; boa constrictors, Boa constrictor; western diamondback rattlesnakes, Crotalus atrox, and savannah monitor lizards, Varanus exanthematicus). Analyses revealed that the isotopic composition of reptile carcasses did not change significantly in response to bouts of starvation lasting up to 168 days. In contrast, the isotopic signatures of reptile excreta became significantly enriched in 15N and depleted in 13C during starvation. The isotopic signatures of reptile scales and lizard claws were less indicative of starvation time than those of excreta. We discuss the physiological mechanisms that might be responsible for the starvation-induced changes in 13C and 15N signatures in the excreta, and present a mixing model to describe the shift in excreted nitrogen source pools (i.e. from a labile source pool to a nonlabile source pool) that apparently occurs during starvation in these animals. The results of this study suggest that naturally occurring stable isotopes might ultimately have some utility for characterizing nitrogen and carbon stress among free-living reptiles.


Visible light-induced crosslinkable gelatin.
A novel visible light-crosslinkable porcine gelatin was prepared for gelation and micropatterning. The preparation employed a photo-oxidation-induced crosslinking mechanism. First, furfuryl groups were incorporated into the gelatin. Second, the modified gelatin was mixed in water with Rose Bengal, which is a visible light sensitizer. Irradiation by visible light solidified the aqueous solution. In addition, when the solution was cast on a plate, dried and photo-irradiated in the presence of a photomask a micropattern was formed that matched the micropattern on the photomask. The gelatin-immobilized regions enhanced cell adhesion. It was also confirmed that the gelatin incorporating furfuryl and Rose Bengal have no significant toxicity. The photo-crosslinkable gelatin was employed as a direct pulp capping material in the dental field. Considering these results, this system could be useful as a new type of visible light-induced crosslinkable biosealant.


Helping patients to give informed consent.
This is the second article in a seven-part series demonstrating the value of analysing patient narratives in nursing and nurse education. Part 1 reviewed the evidence regarding the use of narrative and focused on unintended communication. This article draws on a story exploring consent and capacity, and demonstrates the lessons that can be learnt for all fields of nursing. We hope your reflections on the narratives will help to inform your professional practice and provide evidence for revalidation.


Liver-directed gene therapy: promises, problems and prospects at the turn of the century.
Although liver-directed gene therapy arrived later than gene therapy directed at bone marrow cells, intrinsic advantages of the liver as a target organ make it likely that gene therapy for liver diseases will be among the first therapeutically relevant applications of this treatment modality at the onset of the 21st century. Vectorology for gene transfer to the liver is advancing rapidly, and it is safe to predict that gene therapy vehicles that will be in clinical use a decade from now, have not yet been developed. None of the currently available modes of gene transfer to the liver is optimal for all types of applications. Nonetheless, the concerted effort of many investigators has provided a wide choice of non-viral and viral vectors for gene transfer to the liver for use in specific situations. Original strategies for liver-directed gene therapy included substitution of missing gene products, overexpression of intrinsic or extrinsic genes and inhibition of expression of specific genes. To the list is now added the possibility of site-specific correction or generation of mutations within specific genes in somatic cells of living adult animals. Thus, despite some initial faux pas, liver-directed gene therapy is poised to make an important impact on health care in the year 2000 and beyond.


Knee kinematics in medial osteoarthritis during in vivo weight-bearing activities.
Dynamic knee kinematics were analyzed for medial osteoarthritic (OA) knees in three activities, including two types of maximum knee flexion. Continuous x-ray images of kneeling, squatting, and stair climbing motions were taken using a large flat panel detector. CT-derived bone models were used for the model registration-based 3D kinematic measurements. Three-dimensional joint kinematics and contact locations were determined using two methods: bone-fixed coordinate systems and by interrogation of CT-based bone model surfaces. The femur exhibited gradual external rotation with knee flexion for kneeling and squatting activities, and gradual internal rotation with knee extension for stair climbing. From 100 degrees to 120 degrees flexion, contact locations showed a medial pivot pattern similar to normal knees. However, knees with medial OA displayed a femoral internal rotation bias and less posterior translation when compared with normal knees. A classic screw-home movement was not observed in OA knees near extension. Decreased variability with both activities and methods of calculation were demonstrated for all three activities. In conclusion, the weight-bearing kinematics of patients with medial OA differs from normal knees. Pathological changes of the articulating surfaces and the ligaments correspond to observed abnormalities in knee kinematics.


VP4 and VP7 genotyping of rotavirus samples recovered from infected children in Ireland over a 3-year period.
Between September 1995 and August 1998, the incidence and diversity of the main human rotavirus genotypes (G1, G2, G3, and G4 and P[8], P[4], P[6], and P[9]) among Irish children were determined by using established and adapted reverse transcriptase PCR-based genotyping methods. From a total of 193 rotavirus-positive specimens collected from nine hospitals we successfully identified the P type in 182 (94%) of the samples and the G type in 165 (85.5%) of the samples. Only four samples could not be assigned a G or P type. Two P types existed in Ireland, P[8] (78%) and P[4] (16%), and their relative incidence varied over the 3 years of this study. No P[6] or P[9] types were detected. G1 was the most predominant G type (55%), and the incidences of G2, G3, and G4 isolates were 15.5, 1, and 11%, respectively. Three percent of the samples tested had a mixed G type. A P and G type was assigned to 158 (81.8%) of samples. Of the typeable samples, G1 P[8] was the most prevalent (65%), whereas G2 P[4] (17%), G3 P[8] (1%), G4 P[8] (12%), and mixed types (all G1/ G4 P[8]) (4%) were detected less frequently. In the third year a significant genotypic shift from G1 P[8] to G2 P[4] and G4 P[8] was observed. During the study, we noticed that the inclusion of random primers during cDNA synthesis greatly increased the specificity of the PCR typing assays. No correlation was seen between the contributing hospitals and a specific genotype. In conclusion, the coverage of infection given by the recently licensed tetravalent vaccine would be significantly high in Ireland, although future monitoring of genotypic changes among Irish isolates should be encouraged.


[The new financing system in the Italian National Health Service. Implications for internal medicine].
Since 1995, the Italian National Health Service has begun to fund its hospitals on a DRG basis. This paper presents the main features of the DRG system as well as its likely implications for general internal medicine. The first part describes the introduction of DRGs in the US. The first paragraphs summarize the features of the US health-care system and particularly its private nature with two major exceptions: Medicare and Medicaid. The development of the DRG system and its adoption by Medicare are then described. Finally, the main effects of Medicare's DRG system are underlined: shorter hospital stays, fewer hospital admissions, several diagnostic and surgical procedures shifted from the inpatient to the outpatient setting, and apparently no negative quality implications. The second part focuses on Italy, in general and with specific reference to general internal medicine. For general internal medicine, the new funding system has two major implications. First, it may lead to the creation of larger medical departments including both the current general internal medicine divisions and the various specialties. Second, even under the current organisational structure, divisions will be increasingly required to produce positive financial margins. In this respect, general internal medicine divisions seem to be in a favourable position, especially in terms of costs (both per-diem and per-admission).


Mineral and glycerol concentrations in drinking water on body weight loss and acid-base balance in feed-deprived Holstein bulls.
Situations in which cattle are feed-deprived over extensive periods of time are common in the context of transport and is an animal welfare concern which may also compromise growth and carcass yield grade. This study investigated how the main components of an oral rehydration solution would affect BW loss and blood parameters in feed-deprived bulls. Three dose-response experiments were performed with 24 bulls each (n = 6 per treatment) to study the effect of mineral concentration in Study I (0, 100, 200 and 300 mOsm/kg), the K+ to Na+ ratio in Study II (25:75, 40:60, 60:40 and 75:25), and glycerol concentration in Study III (0%, 1%, 2% and 4% of the final solution). The blocking factor was initial BW and treatments were randomly assigned within each block. Measurements included fluid intakes, BW, and blood parameters at 0, 24 and 48 h relative to the start of feed deprivation. In Study I, increasing mineral concentration in solution linearly decreased BW losses at 48 h. At 24 and 48 h, serum urea linearly decreased with increasing mineral concentration. At 48 h, blood K+ and Na+ linearly increased, resulting in increased blood osmolarity. Additionally, at 24 h feed deprivation, blood pH linearly increased with increasing osmolality. In Study II, BW losses decreased with increasing K+ to Na+ ratio at 24 h, but not at 48 h. No effect of the K+ to Na+ ratio was found on blood parameters, apart from a trend for a linear decrease of blood osmolarity at 48 h. In Study III, serum urea tended to linearly decrease with increasing glycerol inclusion at 24 h, while blood glucose linearly increased with glycerol inclusion at 24 and 48 h. These combined results indicated that a solution with an osmolality of 200 mOsm/kg and a high K+ to Na+ ratio would effectively mitigate BW losses and maintain blood acid-base balance, whereas high glycerol inclusion sustained blood glucose in feed-deprived bulls.


Parathyroid surgery in chronic renal failure.
In hyperparathyroidism associated with endstage renal failure, either subtotal, or total parathyroidectomy with autotransplantation have been advocated as potentially curative for those patients non-responsive to medical therapy. Seventeen such patients managed by the Wellington renal unit were reviewed as to indications for, and responses to, surgery. Itch, psychiatric symptoms, joint ache, muscular weakness, gritty eyes and thirst were the major symptoms for which surgery was recommended. Hypercalcaemia (universal in women) and deterioration in bone radiology were additional indications for operation. All showed remarkable postoperative improvement. Symptomatic hypocalcaemia was significant in 41%. This observation has led to intensified pre- and postoperative vitamin D and calcium therapy. Postoperative radiologic improvement at three months was apparent in 80% of patients. We advocate subtotal parathyroidectomy as the effective surgical treatment of choice for uraemic hyperparathyroidism.


Cytokine overexpression and constitutive NFkappaB in cancer.
The NFkappaB family of transcription factors, central mediators of immune responses, are also involved in oncogenesis. Loss of regulation of the normally latent NFkappaB contributes importantly to the deregulated growth, resistance to apoptosis and propensity to metastasize observed in many cancers. Thus, pathways for activation of NFkappaB are promising targets for new agents that may help to prevent or treat cancer. We find that the abnormal secretion of multiple cytokines that activate NFkappaB by binding to cell-surface receptors is one of the major causes of constitutive NFkappaB activity in cancer. A novel finding is that the latent form of TGFbeta, secreted by some of these cells, can activate NFkappaB. To understand the basis of this abnormal cytokine secretion, we are using forward genetic methods to identify specific causative mutations in cancer cells.


CRISPRroots: on- and off-target assessment of RNA-seq data in CRISPR-Cas9 edited cells.
The CRISPR-Cas9 genome editing tool is used to study genomic variants and gene knockouts, and can be combined with transcriptomic analyses to measure the effects of such alterations on gene expression. But how can one be sure that differential gene expression is due to a successful intended edit and not to an off-target event, without performing an often resource-demanding genome-wide sequencing of the edited cell or strain? To address this question we developed CRISPRroots: CRISPR-Cas9-mediated edits with accompanying RNA-seq data assessed for on-target and off-target sites. Our method combines Cas9 and guide RNA binding properties, gene expression changes, and sequence variants between edited and non-edited cells to discover potential off-targets. Applied on seven public datasets, CRISPRroots identified critical off-target candidates that were overlooked in all of the corresponding previous studies. CRISPRroots is available via https://rth.dk/resources/crispr.


A 1H n.m.r. study of the kinetic properties expressed by glyceraldehyde phosphate dehydrogenase in the intact human erythrocyte.
Glyceraldehyde phosphate dehydrogenase is one of four glycolytic enzymes in the human erythrocyte that together can catalyse exchange of isotope between the C-2 position of lactate and solvent. Detailed measurements of the exchange can be made by using a non-invasive spin-echo p.m.r. method that has been described previously [Brindle, Brown, Campbell, Foxall & Simpson (1982) Biochem. J. 202, 589-602]. By studying the dependence of the exchange on the activity of an individual enzyme, the specific isotope-exchange equilibrium velocity of the enzyme can be measured. Suggestions that glyceraldehyde phosphate dehydrogenase is bound to the membrane in the intact cell, and that it may, under certain conditions, be rate-limiting for glycolytic flux, were examined in the present study by comparing the exchange properties expressed by the enzyme in situ and in vitro. It is concluded that glyceraldehyde phosphate dehydrogenase is not rate-limiting for glycolytic flux and that it is unlikely that a significant fraction of the enzyme is bound to the erythrocyte membrane in situ.


A mutation in MTM1 causes X-Linked myotubular myopathy in Boykin spaniels.
The purpose of this study was to report the findings of clinical and genetic evaluation of a 3-month old male Boykin spaniel (the proband) that presented with progressive weakness. The puppy underwent a physical and neurological examination, serum biochemistry and complete blood cell count, electrophysiological testing, muscle biopsy and whole genome sequencing. Clinical evaluation revealed generalized neuromuscular weakness with tetraparesis and difficulty holding the head up and a dropped jaw. There was diffuse spontaneous activity on electromyography, most severe in the cervical musculature. Nerve conduction studies were normal, the findings were interpreted as consistent with a myopathy. Skeletal muscle was grossly abnormal on biopsy and there were necklace fibers and abnormal triad structure localization on histopathology, consistent with myotubular myopathy. Whole genome sequencing revealed a premature stop codon in exon 13 of MTM1 (ChrX: 118,903,496 C > T, c.1467C>T, p.Arg512X). The puppy was humanely euthanized at 5 months of age. The puppy's dam was heterozygous for the variant, and 3 male puppies from a subsequent litter all of which died by 2 weeks of age were hemizygous for the variant. This naturally occurring mutation in Boykin spaniels causes a severe form of X-linked myotubular myopathy, comparable to the human counterpart.