Three-dimensional computed tomographic volumetric changes in pancreas before and after living donor surgery for pancreas transplantation: effect of volume on glucose metabolism.
In the present study, we aimed to compare the pancreas volumetric changes before and after living donor surgery for pancreas transplantation, using three-dimensional (3D) computed tomography (CT) and glucose metabolism. Pancreatic volume (PV) measurement using 3D CT was performed in 13 consecutive donors who underwent distal pancreatectomy for simultaneous living donor pancreas and kidney transplantation. PV was measured using a workstation before and 3 months after living donor operation. As the parameters of glucose metabolism, hemoglobin A1c (HbA1c) level, fasting plasma glucose (FPG) level, body mass index (BMI), homeostasis model assessment of insulin resistance (HOMA-IR), and insulinogenic index (IGI) were examined simultaneously with the PV measurement. The preoperative and postoperative PVs of pancreas was 30 ± 5 mL and 42 ± 9 mL, respectively. The postoperative PV was significantly higher than the preoperative PV (P < .01) and increased by approximately 40% at 3 months after surgery. The postoperative FPG and HbA1c levels were significantly higher than the preoperative values (P < .01). BMI decreased significantly after surgery (P < .01). No differences in HOMA-IR and IGI were noted between before and after surgery. Diabetes mellitus was not observed any of the 13 living donors during this period. Distal pancreatectomy for living donors caused an increase in the PV and maintained insulin resistance, but it was not sufficient to maintain glucose metabolism at the preoperative state.


Absolute calibration of auroral photometers and ground-based measurements in the 3200-7500-a spectral region.
A convenient laboratory calibration method is outlined. Uncertainties introduced by instrumental and atmospheric effects are discussed. The conclusion is that laboratory calibration is to be preferred. The variable atmospheric extinction can introduce great errors in ground-based measurements, and therefore necessitates separate monitoring. The over-all uncertainty of ground-based measurements can be below 20%.


Cost effectiveness and return on investment of a scalable community weight loss intervention.
This study assessed the lifetime health and economic consequences of an efficacious scalable community weight loss program for overweight and obese adults. We applied a state-transition Markov model to project lifetime economic outcome (US dollar) and the degree of disease averted as a result of a weight loss intervention, compared with no intervention, from a payer perspective. Effect sizes of the intervention on weight loss, by sex, race and ethnicity, and body mass index (BMI) of participants, were derived from a 12-month community program. Relative risk of diseases across BMI levels and other parameters were informed by the literature. A return on investment (ROI) analysis was conducted to present the overall cost-benefit of the program. Simulation results showed that among 33,656 participants and at a cost of $2.88 million, the program was predicted to avert (with a corresponding estimated medical costs saved of) 78 cases of coronary heart disease ($28 million), 9 cases of strokes ($971,832), 92 cases of type 2 diabetes ($24 million), 1 case of colorectal cancer ($357,022), and 3 cases of breast cancer ($483,259) over the participant lifetime. The estimated medical costs saved per participant was $1403 ($1077 of African American men and $1532 of Hispanic men), and the ROI was $16.7 ($12.8 for African American men and $18.3 for Hispanic men) for every $1 invested. We concluded that a scalable efficacious community weight loss program provides a cost-effective approach with significant ROI, which will assist informed decisions for future adoption and dissemination.


Post-brachytherapy initial tumour regression rate correlates with metastatic spread in posterior uveal melanoma.
To confirm the prognostic value of post-brachytherapy initial tumour regression rate in posterior uveal melanoma. A prospective, comparative, observational cohort study was carried out on 147 eyes (147 patients) with choroidal melanoma (mean age 61 years) treated with Ru-106 brachytherapy. Patients were followed clinically and ultrasonically every 6.7 (SD 0.3) months (1001 examinations). On average each patient was examined 5.8 times (mean follow up 9.6 (3.7) years). The echographic parameters included tumour base size, height, internal reflectivity, regularity, vascularity, and extrascleral extension. The clinical follow up included ocular examination and periodic metastatic screening (liver function tests and liver imaging). Main outcome measures were risk of liver metastasis in correlation with the post-brachytherapy initial tumour regression rate. At brachytherapy the mean tumour height was 5.2 mm (range 2.2-11.8 mm). After brachytherapy 142 tumours (96.6%) responded by a decrease in height. The initial height regression rate was 6.1% (0.8%) per month in patients who later developed metastasis v 4.3% (0.4%) per month in those who did not. Tumours higher than 6 mm, tumours with an internal reflectivity smaller than 50%, and tumours with an initial rate of height regression larger than 0.7 mm/month (10% per month) had higher 5 year melanoma related mortality. Kaplan-Meier survival analysis and the multivariate Cox proportional hazards model showed a significant role for tumour height and initial tumour regression rate on patients' survival. This study confirms that post-brachytherapy initial tumour regression rate has a prognostic value.


Relationship between the body mass index and the ponderal index with physical fitness in adolescent students.
The relationship between the Body Mass Index (BMI) with physical fitness in children and adolescent populations from diverse regions are consistent. However, the relationship between the Ponderal Index (PI) with physical fitness, based on what is known to date, has not been examined in depth. The objective was to evaluate the relationships between BMI and PI with three physical fitness tests of students living at moderate altitudes in Peru. A descriptive correlational study was carried out with 385 adolescents, between the ages of 10.0 to 15.9 years old, from the province of Arequipa, Peru. Weight, height, and three physical fitness tests (horizontal jump, agility, and abdominal muscle resistance) were evaluated. BMI and PI were calculated, and they were, then, categorized into three strata (low, normal, and excessive weight). Specific regressions were calculated for sex, using a non-lineal quadratic model for each item adjusted for BMI and PI. The relationship between BMI and PI with the physical tests reflected parabolic curves that varied in both sexes. The regression values for BMI in males oscillated between R2 = 0.029 and 0.073 and for females between R2 = 0.008 and 0.091. For PI, for males, it varied from R2 = 0.044 to 0.82 and for females, from R2 = 0.011 to 0.103. No differences occurred between the three nutritional categories for BMI as well as for PI for both sexes (p range between 0.18 to 0.38), as well as for low weight (BMI vs PI), normal weight (BMI vs PI), and excessive weight (BMI vs PI) (p range between 0.35 to 0.64). BMI showed inferior quadratic regressions with respect to the PI. In addition, physical performance was slightly unfavorable when it was analyzed by BMI. PI could be a useful tool for analyzing and predicting physical fitness for adolescents living at a moderate altitude since it corrects for the notable differences for weight between adolescents.


Evaluation of the enterotube system for identification of members of the family Enterobacteriaceae.
The Enterotube system was evaluated, in parallel with conventional bacteriological procedures for the identification of members of the family Enterobacteriaceae, by using bacterial strains from a variety of clinical specimens and from stock cultures. Excellent agreement between the two test systems was obtained with the following reactions: hydrogen sulfide, indole, Simmons' citrate, glucose, and lactose. Agreement was not as good (<85%) with the urea, phenylalanine deaminase, and dulcitol reactions. The Enterotube lysine decarboxylase test was unsatisfactory. The Enterotube method will correctly identify strains of the family Enterobacteriaceae approximately 50% of the time; if identification only as Klebsiella-Enterobacter-Serratia group is needed, the method will be correct 85% of the time. On the basis of this evaluation, the Enterotube system appears to be both simple and rapid for the presumptive identification of these bacteria. Because of the limited usefulness of the lysine decarboxylase test, the results obtained by this test system are less reliable than those obtained by conventional methods.


Faster V̇O2 kinetics after priming exercises of different duration but same fatigue.
This study compared the responses of two priming exercises of similar fatigue on the adjustment of the oxygen uptake time constant (τV̇O2) in cycling. Ten healthy young adults (25 ± 3 yr) performed: three step transitions from a 20-W baseline to the power output (PO) below the gas exchange threshold (MOD, MODPRE); a 3-min bout (P3MIN) at 90% of peak PO (POpeak), followed by MOD (MOD3MIN); and a 6-min bout (P6MIN) at 80% of POpeak, followed by MOD (MOD6MIN). The O2 supply-to-O2 demand ([HHb]/V̇O2) ratio was calculated for MODPRE, MOD3MIN, and MOD6MIN. Neuromuscular fatigue was measured isometrically pre- and post-priming exercise. Reductions in maximal voluntary contraction (-29 ± 6 vs -34 ± 7%) and high-frequency doublet amplitude (-48 ± 13 vs -43 ± 11%) were not significantly different between P3MIN vs P6MIN, suggesting similar fatigue. τV̇O2 for MOD3MIN and MOD6MIN were similar, being ~25% smaller than MODPRE. The [HHb]/V̇O2 ratio was significantly greater in MODPRE (1.13 ± 0.12) compared to MOD3MIN (1.02 ± 0.04) and MOD6MIN (1.02 ± 0.04). This study showed that priming exercise of shorter duration and higher intensity, was sufficient to accelerate V̇O2 kinetics similarly to that observed subsequent to P6MIN when the muscle fatigue was similar.


Methicillin-resistant staphylococcal bacteremia in patients with hematologic malignancies: clinical and microbiological retrospective comparative analysis of S. haemolyticus, S. epidermidis and S. aureus.
Although Staphylococcus haemolyticus (SH) represents an emerging etiology of methicillin-resistant (MR) coagulase-negative staphylococcal nosocomial bacteremia, little is known of clinical significance of this infection. Thus, we performed case-control retrospective comparative analysis of MRSH bacteremias (MRSHB), methicillin-resistant S. epidermidis bacteremias (MRSEB), and methicillin-resistant S. aureus bacteremias (MRSAB) in patients with hematologic malignancies. Most patients in the three groups were neutropenic and had a central venous catheter (CVC) in place at the onset of bacteremia. However, MRSHB patients had a CVC in place prior to bacteremia for a time significantly more prolonged than MRSEB and MRSAB ones (p<0.05). Severe sepsis or septic shock were more common in patients with MRSAB as compared with MRSHB (p=0.02). Nosocomial attributable mortality rate was very low in the 3 study groups (0 to 5.4%) and only two patients developed metastatic infections. Overall, reduced susceptibility to teicoplanin was observed in 19 (47.5%) MRSH and in 4 (10%) MRSE isolates. Resistance to teicoplanin was observed in 6 isolates, all MRSH. Reduced susceptibility or resistance to vancomycin was observed in 2 isolates, both MRSH. All MRSA isolates were susceptible to the glycopeptides. Comparison between cases of bacteremia in patients with MRSH isolates with reduced susceptibility to teicoplanin and those with susceptible MRSH did not reveal significant differences in the clinical-microbiological response to teicoplanin therapy and outcome. Our results seem to suggest that MRSHB in hematologic patients is associated with low morbidity and mortality rates. MRSH frequently shows a reduced susceptibility to teicoplanin; however these in vitro data do not seem associated with an unfavorable clinical response to teicoplanin therapy for MRSHB in patients with hematologic malignancies.


Barriers to medical help-seeking among older men with prostate cancer.
Men's disinclination to seek medical help has been linked to higher rates of morbidity and mortality compared to women. However, previous studies were conducted predominantly with healthy, young, and middle-aged men. We explored the perceived medical barriers to help-seeking in older men with prostate cancer. 20 men with prostate cancer took part in semistructured interviews, which were analyzed using thematic analysis. Three themes were identified related to negative attitudes toward help-seeking: male gender role; fear of the health condition, medical and treatment procedures; and embarrassment as a consequence of medical examinations, communication with health (and nonhealth) professionals, and the disclosure of sexual-related symptoms. The barriers identified in our study strengthen the evidence for the impact of traditional masculinity on help-seeking in men.


Preventing alcohol use among late adolescent urban youth: 6-year results from a computer-based intervention.
This study was undertaken to evaluate the efficacy of a skills-based CD-ROM intervention, with and without a parent component, to reduce alcohol use among urban youth at 6-year follow-up. At recruitment, 513 youths with a mean age of 10.8 years were randomly assigned to one of three study arms: youth CD-ROM intervention plus parent component, youth CD-ROM intervention only, or control. All youths completed pretest, posttest, and annual follow-up measures. Youths and parents in their respective arms received the initial intervention program between pretest and posttest measures and received booster interventions between each follow-up measure. With 80% sample retention at 6-year follow-up, youths in both intervention arms reported less past-month alcohol and cigarette use and fewer instances of heavy drinking and negative alcohol-related consequences. Despite having similar numbers of drinking peers as youths in the control arm, youths in both intervention arms reported greater alcohol-refusal skills. Only past-month cigarette use differed between the two intervention arms, with youths in the intervention-plus-parent-component arm smoking less than youths in the CD-ROM intervention-only arm. Six years after initial intervention, youths who received a culturally tailored, skills-based prevention program had reduced alcohol use and lower rates of related risky behaviors than youths in the control arm.


Microvessel hematocrit: measurement and implications for capillary oxygen transport.
Microvascular preparations of the hamster cheek pouch and cremaster muscle were used to establish the relationship among three measures of red cell distribution: 1) the systemic hematocrit defined as the volume percentage of red cells sampled from a toe clip; 2) the tube hematocrit defined as the instantaneous volume fraction of red cells in a microvessel segment of measured length and diameter; and 3) the discharge hematocrit defined as the volume percentage of red cells that flowed into micropipettes inserted into microvessels. The results show the tube hematocrit is low and highly variable in the presence of a nearly constant systemic hematocrit. The discharge hematocrit, in contrast, consistently approximated systemic values in blood taken from arterioles or venules with diameters of 6-98 micron. Indeed, the mean ratio of the discharge to the systemic hematocrit did not differ from unity (P greater than 0.25). The observed similarity between the discharge and systemic hematocrit indicates that red cells are distributed uniformly across capillary networks. Differences between the absolute value and variance of the tube hematocrit compared with the discharge hematocrit imply that the anatomical volume of a microvessel can differ from the volume available to cells and plasma within a microvessel. Thus moment-to-moment variation in the tube hematocrit or capillary red cell spacing may be indicative of a change in the effective capillary flow cross section and not solely on the inflow hematocrit.


Non-invasive evaluation of the myocardial substrate of cardiac amyloidosis by gadolinium cardiac magnetic resonance.
To investigate the prevalence and distribution of gadolinium (Gd) enhancement at cardiac magnetic resonance (CMR) imaging in patients with cardiac amyloidosis (CA) and to look for associations with clinical, morphological, and functional features. 21 patients with definitely diagnosed CA (nine with immunoglobulin light chain amyloidosis and 12 transthyretin related) underwent Gd-CMR. Gd enhancement was detected in 16 of 21 (76%) patients. Sixty six of 357 (18%) segments were enhanced, more often at the mid ventricular level. Transmural extension of enhancement within each patient significantly correlated with left ventricular (LV) end systolic volume (r = 0.58). The number of enhanced segments correlated with LV end diastolic volume (r = 0.76), end systolic volume (r = 0.6), and left atrial size (r = 0.56). Segments with > 50% extensive transmural enhancement more often were severely hypokinetic or akinetic (p = 0.001). Patients with > 2 enhanced segments had significantly lower 12 lead QRS voltage and Sokolow-Lyon index. No relation was apparent with any other clinical, morphological, functional, or histological characteristics. Gd enhancement is common but not universally present in CA, probably due to expansion of infiltrated interstitium. The segmental and transmural distribution of the enhancement is highly variable, and mid-ventricular regions are more often involved. Enhancement appears to be associated with impaired segmental and global contractility and a larger atrium.


Intraoperative Simulation: Team Training for Resuscitation While Using Intraoperative Computed Tomography: 2-Dimensional Operative Video.
Spine surgeons increasingly use intraoperative computed tomography (iCT) to facilitate surgery. iCT has several advantages, including the ability to decrease radiation exposure, improve surgical accuracy, and decrease operative time.1-3 However, the large footprint of the equipment can impede fast patient access in the event of an emergency resuscitation. This challenge is compounded when the patient is prone with rigid head fixation. To achieve fast, high-quality resuscitation, a large team must overcome numerous challenges. Cohesive team functioning under these circumstances requires planning, practice, and refinement.4 As a result of our simulation sessions, we have made several changes to the setup of our iCT cases. The following equipment is now routinely used: extralong tubing between the anesthesia circuit and patient, portable vital monitor, additional intravenous access is obtained, and extension tubing is used with all lines. We have created educational diagrams to streamline 2 challenging processes: optimal bed placement (for supination) and removal of equipment from the operating room (OR) to accommodate an influx of emergency personnel and equipment. Since the implementation of this protocol, 1 prone posterior cervical patient had intraoperative cardiac arrest. The protocol was followed. Return of spontaneous circulation was achieved within 5 min. The patient was discharged from the hospital with no neurological sequelae. During debriefing, stakeholders uniformly credited the simulated practice with this positive outcome. Emergency planning is a multifaceted process that continually evolves. With a steady flux of personnel and equipment, ongoing practice is essential to ensure readiness. Here, we share the key elements of our twice-yearly simulation. This simulation was performed on a training mannequin. This study did not involve human subjects. Any depictions of care rendered to nonidentifiable patients were standard (nonexperimental).


Comparison of standard methods and gas chromatography method in determination of formaldehyde emission from MDF bonded with formaldehyde-based resins.
Formaldehyde emissions from MDF bonded with urea-formaldehyde resin (UF), melamine-formaldehyde resin (MF) and the co-polycondensation resin of urea-melamine-formaldehyde (UMF) and melamine-formaldehyde, measured by the Japanese standard method of determining formaldehyde emission with a desiccator (JIS A 5908) and the DIN EN 120 (European Committee For Standardization, 1991) method using the perforator value, were used as the typical standard methods. While the UF resin showed a desiccator value of 7.05 ppm and a perforator value of 12.1 mg/100 g panel, the MF resin exhibited a desiccator value of 0.6 ppm and a perforator value of 2.88 mg/100 g panel. According to the Japanese industrial standard and the European standard, the formaldehyde emission level of the MDF panels made with UF resin in this study was E(2) grade. The formaldehyde emission level was dramatically reduced by the addition of MF resin. This is because the addition of formaldehyde to melamine occurs more easily and completely than its addition to urea, even though the condensation reaction of melamine with formaldehyde is similar to that between urea and formaldehyde. These two methods, the desiccator method and the perforator method, produced proportionally equivalent results. Gas chromatography, a more sensitive and advanced method, was also used. The samples used for gas chromatography were gathered during the experiment involving the perforator method. The formaldehyde emission levels obtained from gas chromatography were similar to those obtained from the perforator method. The formaldehyde contents measured by gas chromatography were directly proportional to the perforator values.


A critical period of protein kinase activity after tetanic stimulation is required for the induction of long-term potentiation.
A critical period of protein kinase activity required for the induction of long-term potentiation (LTP) was determined in area CA1 or hippocampal slices using the broad-range and potent protein kinase inhibitors K-252a and staurosporine. As reported previously, K-252a and staurosporine blocked LTP induction when applied before, during, and after high-frequency stimulation (HFS). In contrast, K-252a did not block LTP when applied only before and during HFS and washed out immediately after HFS. K-252a and staurosporine both attenuated LTP magnitude when applied immediately after or as late as 5 min after HFS. However, K-252a applications beginning 30-45 min after HFS did not affect LTP expression significantly. K-252a had no detectable effect on isolated N-methyl-D-aspartate (NMDA) receptor-mediated EPSPs but significantly inhibited the in situ phosphorylation of specific hippocampal proteins (synapsin I, MARCKS, and B-50). In addition, K-252a attenuated 4 beta-phorbol-12,13-dibutyrate (PDBu)-enhanced synaptic transmission. Our results indicate that there is a critical period of protein kinase activity required for LTP induction that extends for approximately 20 min after HFS. In addition, our results suggest that protein kinase activity during and immediately after HFS is not sufficient for LTP induction. These results provide new information about the mechanisms that underlie LTP induction and expression and provide evidence for persistent and/or Ca(2+)-independent protein kinase activity involvement in LTP.


Histopathological features of the Harderian glands in transgenic mice carrying MMTV/N-ras protooncogene.
The Harderian gland is a tubule-alveolar gland found within the orbit, on the posterior aspect of the eyeball. In mice, it is composed almost exclusively of secretory tubules and alveoli. The Harderian glands of transgenic mice, carrying the activated N-ras oncogene under the transcriptional control of the mouse mammary tumor virus long terminal repeat promoter (MMTV-LTR), were examined and compared to those of normal mice. Thirty transgenic mice provided by A. Pellicer (NYU) and 30 normal mice were examined in this study. Harderian glands were dissected, immersed in a formalin-based fixative, and embedded in paraffin. The sections of these glands were studied via histological techniques. Our results show that proliferative alterations in the Harderian glands of these transgenic mice are present even in the youngest animals. Such alterations correspond to different tumoral evolution stages, ranging from hyperplasia to wide tissue destruction. In the most advanced situations, these changes are accompanied by a glandular hypertrophy. Our results suggest a very high tumoral incidence in the Harderian glands of transgenic mice compared to normal mice. Tumors appear spontaneously in some areas, but not at the same time in the whole gland.


In vitro activity of human-simulated epithelial lining fluid exposures of ceftaroline, ceftriaxone, and vancomycin against methicillin-susceptible and -resistant Staphylococcus aureus.
Staphylococcus aureus, including methicillin-susceptible (MSSA) and -resistant (MRSA) strains, is an important pathogen of bacterial pneumonia. As antibiotic concentrations at the site of infection are responsible for killing, we investigated the activity of human-simulated epithelial lining fluid (ELF) exposures of three antibiotics (ceftaroline, ceftriaxone, and vancomycin) commonly used for treatment of S. aureus pneumonia. An in vitro pharmacodynamic model was used to simulate ELF exposures of vancomycin (1 g every 12 h [q12h]), ceftaroline (600 mg q12h and q8h), and ceftriaxone (2 g q24h and q12h). Four S. aureus isolates (2 MSSA and 2 MRSA) were evaluated over 72 h with a starting inoculum of ∼ 10(6) CFU/ml. Time-kill curves were constructed, and microbiological response (change in log10 CFU/ml from 0 h and the area under the bacterial killing and regrowth curve [AUBC]) was assessed in duplicate. The change in 72-h log10 CFU/ml was largest for ceftaroline q8h (reductions of >3 log10 CFU/ml against all strains). This regimen also achieved the lowest AUBC against all organisms (P < 0.05). Vancomycin produced reliable bacterial reductions of 0.9 to 3.3 log10 CFU/ml, while the activity of ceftaroline q12h was more variable (reductions of 0.2 to 2.3 log10 CFU/ml against 3 of 4 strains). Both regimens of ceftriaxone were poorly active against MSSA tested (0.1 reduction to a 1.8-log10 CFU/ml increase). Against these S. aureus isolates, ELF exposures of ceftaroline 600 mg q8h exhibited improved antibacterial activity compared with ceftaroline 600 mg q12h and vancomycin, and therefore, this q8h regimen deserves further evaluation for the treatment of bacterial pneumonia. These data also suggest that ceftriaxone should be avoided for S. aureus pneumonia.


Mimicking the Kidney: A Key Role in Organ-on-Chip Development.
Pharmaceutical drug screening and research into diseases call for significant improvement in the effectiveness of current in vitro models. Better models would reduce the likelihood of costly failures at later drug development stages, while limiting or possibly even avoiding the use of animal models. In this regard, promising advances have recently been made by the so-called "organ-on-chip" (OOC) technology. By combining cell culture with microfluidics, biomedical researchers have started to develop microengineered models of the functional units of human organs. With the capacity to mimic physiological microenvironments and vascular perfusion, OOC devices allow the reproduction of tissue- and organ-level functions. When considering drug testing, nephrotoxicity is a major cause of attrition during pre-clinical, clinical, and post-approval stages. Renal toxicity accounts for 19% of total dropouts during phase III drug evaluation-more than half the drugs abandoned because of safety concerns. Mimicking the functional unit of the kidney, namely the nephron, is therefore a crucial objective. Here we provide an extensive review of the studies focused on the development of a nephron-on-chip device.


Artificial chromosome-based transgenes in the study of genome function.
The transfer of large DNA fragments to the mouse genome in the form of bacterial, yeast or phage artificial chromosomes is an important process in the definition of transcription units, the modeling of inherited disease states, the dissection of candidate regions identified by linkage analysis and the construction of in vivo reporter genes. However, as with small recombinant transgenes, the transferred sequences are usually integrated randomly often with accompanying genomic alterations and variable expression of the introduced genes due to the site of integration and/or copy number. Therefore, alternative methods of integrating large genomic transgenes into the genome have been developed to avoid the variables associated with random integration. This review encourages the reader to imagine the large variety of applications where artificial chromosome transgenes can facilitate in vivo and ex vivo studies in the mouse and provides a context for making the necessary decisions regarding the specifics of experimental design.


The potential role of myocardial serotonin receptor 2B expression in canine dilated cardiomyopathy.
Serotonin signalling in the heart is mediated by receptor subtype 2B (5-HTR2B). A contribution of serotonin to valvular disease has been reported, but myocardial expression of 5-HTR2B and its role in canine dilated cardiomyopathy (DCM) is not known. The aim of the present study was to investigate myocardial 5-HTR2B mRNA expression in dogs with DCM and to correlate results with expression of markers for inflammation and remodelling. Myocardial samples from eight healthy dogs, four dogs with DCM, five with cardiac diseases other than DCM and six with systemic non-cardiac diseases were investigated for 5-HTR2B mRNA expression using quantitative PCR (qPCR). The results were compared to mRNA expression of selected cytokines, matrix metalloproteinases (MMP) and tissue inhibitors of matrix metalloproteinase (TIMP). Laser microdissection with subsequent qPCR and immunohistochemistry were employed to identify the cells expressing 5-HTR2B. The myocardium of control dogs showed constitutive 5-HTR2B mRNA expression. In dogs with DCM, 5-HTR2B mRNA values were significantly greater than in all other groups, with highest levels of expression in the left ventricle and right atrium. Myocytes were identified as the source of 5-HTR2B mRNA and protein. A significant positive correlation of 5-HTR2B mRNA with expression of several cytokines, MMPs and TIMPs was observed. The findings suggest that serotonin might play a role in normal cardiac structure and function and could contribute to myocardial remodelling and functional impairment in dogs with DCM.


Conditional ablation of macrophages disrupts ovarian vasculature.
Macrophages are the most abundant immune cell within the ovary. Their dynamic distribution throughout the ovarian cycle and heterogenic array of functions suggest the involvement in various ovarian processes, but their functional role has yet to be fully established. The aim was to induce conditional macrophage ablation to elucidate the putative role of macrophages in maintaining the integrity of ovarian vasculature. Using the CD11b-diphtheria toxin receptor (DTR) mouse, in which expression of human DTR is under the control of the macrophage-specific promoter sequence CD11b, ovarian macrophages were specifically ablated in adult females by injections of diphtheria toxin (DT). CD11b-DTR mice were given DT treatment or vehicle and ovaries collected at 2, 8, 16, 24 and 48 h. Histochemical stains were employed to characterise morphological changes, immunohistochemistry for F4/80 to identify macrophages and the endothelial cell marker CD31 used to quantify vascular changes. In normal ovaries, macrophages were detected in corpora lutea and in the theca layer of healthy and atretic follicles. As macrophage ablation progressed, increasing amounts of ovarian haemorrhage were observed affecting both luteal and thecal tissue associated with significant endothelial cell depletion, increased erythrocyte accumulation and increased follicular atresia by 16 h. These events were followed by necrosis and profound structural damage. Changes were limited to the ovary, as DT treatment does not disrupt the vasculature of other tissues likely reflecting the unique cyclical nature of the ovarian vasculature and heterogeneity between macrophages within different tissues. These results show that macrophages play a critical role in maintaining ovarian vascular integrity.


Nitric oxide. II. Nitric oxide protects in intestinal inflammation.
This article examines the evidence for nitric oxide (NO) as a protective agent in splanchnic ischemia-reperfusion and other forms of acute intestinal inflammation. Four major points emerge from this body of data. First, acute intestinal inflammation results in an early (i.e., <5 min) and severe decrease in endothelium-derived NO. Thus the early trigger event in this condition is a functional loss of NO. Second, administration of exogenous NO, NO donors, or NO precursors ameliorate splanchnic ischemia-reperfusion and other forms of acute intestinal inflammation (i.e., splanchnic trauma). These beneficial effects occur at physiological levels of NO when given early in the course of the inflammatory state. Third, blockade of nitric oxide synthase (NOS) or gene deletion of NOS exacerbates intestinal inflammation. Fourth, there are a variety of signaling mechanisms that may mediate the protective effect of NO.


COMBINATION OF COMPUTED TOMOGRAPHIC IMAGING CHARACTERISTICS OF MEDIAL RETROPHARYNGEAL LYMPH NODES AND NASAL PASSAGES AIDS DISCRIMINATION BETWEEN RHINITIS AND NEOPLASIA IN CATS.
Feline nasal diseases are a diagnostic challenge. The objective of this retrospective, cross-sectional study was to determine whether computed tomography (CT) imaging characteristics of the medial retropharyngeal lymph nodes (MRPLN), alone or in combination with CT imaging characteristics of the nasal passages, could aid in differentiation between rhinitis and nasal neoplasia. Cats were recruited from record archives at two veterinary facilities during the period of 2008-2012. Selection criteria were presentation for chronic nasal discharge, contrast-enhanced CT of the head that included the MRPLN, and rhinoscopic nasal biopsy resulting in diagnosis of rhinitis or neoplasia. For each CT scan, two board-certified veterinary radiologists recorded MRPLN size, attenuation, heterogeneity, contrast-medium enhancement, margination, shape, presence of a lymph node hilus, perinodal fat, turbinate lysis, paranasal bone lysis, and nasal mass. Both readers were unaware of patient information at the time of CT interpretation. Thirty-four cats with rhinitis and 22 cats with neoplasia were included. Computed tomographic characteristics significantly associated with neoplasia included abnormal MRPLN hilus (OR 5.1), paranasal bone lysis (OR 5.6), turbinate lysis (5.6), mass (OR 26.1), MRPLN height asymmetry (OR 4.5), and decreased MRPLN precontrast heterogeneity (OR 7.0). The combined features predictive of neoplasia were a nasal mass with abnormal hilus (OR 47.7); lysis of turbinates/paranasal bones with abnormal MRPLN hilus (OR 16.2). Findings supported the hypothesis that combining CT features of the nasal passages and MRPLN aided in differentiating rhinitis from neoplasia in cats.


The real catecholamine content of secretory vesicles in the CNS revealed by electrochemical cytometry.
Resolution of synaptic vesicle neurotransmitter content has mostly been limited to the study of stimulated release in cultured cell systems, and it has been controversial as to whether synaptic vesicle transmitter levels are saturated in vivo. We use electrochemical cytometry to count dopamine molecules in individual synaptic vesicles in populations directly sampled from brain tissue. Vesicles from the striatum yield an average of 33,000 dopamine molecules per vesicle, an amount considerably greater than typically measured during quantal release at cultured neurons. Vesicular content was markedly increased by L-DOPA or decreased by reserpine in a time-dependent manner in response to in vivo administration of drugs known to alter dopamine release. We investigated the effects of the psychostimulant amphetamine on vesicle content, finding that vesicular transmitter is rapidly depleted by 50% following in vivo administration, supporting the "weak base hypothesis" that amphetamine reduces synaptic vesicle transmitter and quantal size.


Effect of 12-Week Whole-Body Vibration Exercise on Lumbopelvic Proprioception and Pain Control in Young Adults with Nonspecific Low Back Pain.
BACKGROUND Nonspecific low back pain (NSLBP) accounts for a large proportion of low back pain cases. The present study aimed to investigate the effect of the whole-body vibration (WBV) exercise on lumbar proprioception in NSLBP patients. It was hypothesized that WBV exercise enhances lumbar proprioception. MATERIAL AND METHODS Forty-two patients with NSLBP performed an exercise program 3 times a week for a total of 12 weeks of WBV. The lumbar proprioception was measured by joint position sense. Outcomes were lumbar angle deviation and visual analogue scale (VAS) score. RESULTS After the 12-week WBV exercise, lumbar flexion angle deviation was reduced from 3.65±2.26° to 1.90±1.07° (P=0.0001), and extension angle deviation was reduced from 3.06±1.85° to 1.61±0.75° (P=0.0001), significantly lower than baseline. After participating in the 12-week WBV exercise, a significant pain reduction was observed (P=0.0001). Men in the whole group (n=32) indicated significantly lower angle deviations in flexion and extension, whereas women (n=10) indicated significantly lower flexion angle deviation (P=0.037), and no significant difference was found in extension angle deviation (P=0.052). However, by subdividing the entire group (n=42) into poor and good proprioceptive groups, WBV exercise presented significant enhancement of lumbar proprioceptive ability in the poor flexion proprioception subgroup, poor extension proprioception subgroup, and good extension proprioception subgroup (each P=0.0001), but not in the subgroup with good flexion proprioceptive ability (P=0.165). CONCLUSIONS Lumbar flexion and extension proprioception as measured by joint position sense was significantly enhanced and pain was significantly reduced after 12-week WBV exercise in NSLBP patients. However, the patients with good flexion proprioceptive ability had limited proprioceptive enhancement.


Process Variability-Technological Challenge and Design Issue for Nanoscale Devices.
Current advanced transistor architectures, such as FinFETs and (stacked) nanowires and nanosheets, employ truly three-dimensional architectures. Already for aggressively scaled bulk transistors, both statistical and systematic process variations have critically influenced device and circuit performance. Three-dimensional device architectures make the control and optimization of the device geometries even more important, both in view of the nominal electrical performance to be achieved and its variations. In turn, it is essential to accurately simulate the device geometry and its impact on the device properties, including the effect caused by non-idealized processes which are subject to various kinds of systematic variations induced by process equipment. In this paper, the hierarchical simulation system developed in the SUPERAID7 project to study the impact of variations from equipment to circuit level is presented. The software system consists of a combination of existing commercial and newly developed tools. As the paper focuses on technological challenges, especially issues resulting from the structuring processes needed to generate the three-dimensional device architectures are discussed. The feasibility of a full simulation of the impact of relevant systematic and stochastic variations on advanced devices and circuits is demonstrated.


Lactose quantification in bovine milk by nuclear magnetic resonance without deuterated solvent (No-D qNMR).
Milk is a homogeneous mixture of substances such as lactose, proteins, and glycerides. Among carbohydrates, lactose is a disaccharide composed of glucose and galactose, and it is present in bovine milk at a level of 4.6%. According to resolution no. 135 of the National Health Surveillance Agency (ANVISA) from Brazil, dairy products labeled "lactose-free" must contain 1.0 mg mL-1 or less of this disaccharide. Thus, this work aims to develop and validate a method for quantifying the lactose content by quantitative nuclear magnetic resonance without the use of deuterated solvent (No-D qMNR). The validation of the developed method followed the norms provided by ANVISA resolution RDC no. 166, based on the figures of merit such as selectivity, linearity, the limit of detection (LOD) and quantification (LOQ), accuracy, precision, and robustness. The obtained results validated the method due to excellent linearity, demonstrated by the value of R > 0.990 and the homoscedasticity of the results, as well as precision, accuracy, and robustness values lower than 5%. Furthermore, LOD and LOQ values around 0.1345 mg mL-1 and 0.4076 mg mL-1, respectively, were obtained, which are lower than those required by legislation. The No-D qNMR technique was also able to quantify lactose content in commercial lactose-free milk.


High RNA-Binding Motif Protein 3 (RBM3) Expression is Independently Associated with Prolonged Overall Survival in Intestinal-Type Gastric Cancer.
BACKGROUND High expression of the RNA-binding motif protein 3 (RBM3) has previously been described as a favorable clinicopathological factor in several cancers, including ovarian cancer, colorectal cancer, prostate cancer, and breast cancer. The aim of this study was to examine the prognostic implications of RBM3 expression in gastric cancer. MATERIAL AND METHODS Immunohistochemical analysis of RBM3 expression from 123 patients showed that upregulated RBM3 was mainly found in intestinal-type (n=78, case=59) cancer compared to diffuse-type (n=15, case=8) and mixed-type (n=30, case=17). There were no significant differences in RBM3 expression in subgroups of clinicopathological parameters. RBM3 expression was strongly associated with p53 but not with Ki-67. Cox univariate analysis revealed that high RBM3 expression was closely associated with prolonged overall survival time (HR 0.504, 95% CI: 0.300-0.845, P=0.009). Multivariate analysis remained supporting these associations when adjusted for age, sex, tumor size, differentiation grade, TNM stage, lymphatic invasion, and Ki-67 and p53 expression (HR 0.541, 95% CI: 0.308-0.952, P=0.033), where Lauren grade was not included. Lauren grade was the only factor with independent prognostic significance in a model adjusted for all factors. These results were confirmed by Kaplan-Meier analysis. RESULTS Therefore, together with the upregulated RBM3 expression observed in intestinal-type of Lauren grade, we suggest that upregulation of RBM3 is partially responsible for the favorable overall survival in cases with intestinal Lauren grade, which is demonstrated by the box diagram and Kaplan-Meier analysis. Our results showed that high RBM3 expression in gastric cancer is mainly found in intestinal-type of Lauren grade and is associated with longer overall survival time. CONCLUSIONS We found that RBM3 is a potential biomarker of good prognosis and deserves further validation.


[Obstructive sleep apnea syndrome and hypertension. Evidence of their relationship].
The purpose of this review is to analyze the relation between obstructive sleep apnea and hypertension. We present epidemiological data of the respiratory disorder and its association with high blood pressure, as well as physiopathological interactions between both conditions, the diagnostic methods, and the impact of treatment on pathophysiology and prognosis.


LINC01121 Inhibits Cell Apoptosis While Facilitating Proliferation, Migration, and Invasion Though Negative Regulation of the Camp/PKA Signaling Pathway via GLP1R.
Pancreatic cancer is an aggressive malignancy as a result of highly metastatic potential. The current study was carried out to alter the expression of LINC01121 in pancreatic cancer, with the aim of elucidating its effects on the biological processes of cell proliferation, migration, invasion, and apoptosis. We hypothesized that both the GLP1R gene and cAMP/PKA signaling pathway participate in the aforementioned process. Microarray data (GSE14245, GSE27890 and GSE16515) and annotating probe files linked to pancreatic cancer were downloaded through the GEO database. The Multi Experiment Matrix (MEM) site was used to predict the target gene of lncRNA. Both pancreatic cancer tissues (n = 56) and paracancerous tissues (n = 45) were collected from patients diagnosed with pancreatic cancer. Immunohistochemistry was applied to identify the positive expression rate of GLP1R protein. Isolated pancreatic cancer cells and PANC-1 cells were independently classified into the blank, negative control (NC), LINC01121 vector, siRNA-LINC01121, siRNA-GLP1R and siRNA-LINC01121 + siRNA-GLP1R groups. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis were applied to detect the expressions of LINC01121, GLP1R, cAMP, PKA, CREB, Bcl-2, Bad and PCNA. Cell proliferation, migration, invasion, cycle progression, and apoptosis were examined by MTT assay, scratch test, Transwell assay and flow cytometry analyses of Annexin V-FITC/PI staining. Observations were made indicating that LINC01121 was highly expressed, while low expressions of GLP1R in pancreatic cancer were detected based on microarray data, which was largely in consistent with the data collected of LINC01121 and GLP1R within the tissues. The target prediction program and luciferase activity analysis was testament to the notion suggesting that GLP1R was indeed a target of LINC01121. In contrast to the blank and NC groups, the LINC01121 vector group exhibited increased expressions of LINC01121; decreased mRNA and protein levels of GLP1R, Bad, cAMP, and PKA; increased protein levels of CREB, Bcl-2, PCNA, p-PKA and p-CREB; increased cell proliferation, migration and invasion; and decreased cell apoptosis. There was no significant difference detected among the blank, NC, and siRNA-LINC01121 + siRNA-GLP1R groups, except that decreased LINC01121 expression was determined in the siRNA-LINC01121 + siRNA-GLP1R group. Parallel data were observed in the pancreatic cancer cells and PANC-1 cells. The current study presents evidence indicating that LINC01121 might inhibit apoptosis while acting to promote proliferation, migration, and invasion of pancreatic cancer cells, supplementing the stance held that LINC01121 functions as a tumor promoter by means of its involvement in the process of translational repression of the GLP1R and inhibition of the cAMP/PKA signaling pathway.


Medication Knowledge Among Older Adults Admitted to Home Care in Ontario During 2012-2013.
Medication use among Canadian seniors is widespread and increases with the number of comorbidities. Limited evidence exists on medication knowledge among seniors, especially in home care. The purpose of this retrospective cohort study was to describe medication knowledge and ability to take medication among seniors admitted to home care in Ontario. Ten percent had little or no knowledge of what medication to take (n = 1,389/14,004) or an understanding of the purpose of their medications (n = 1,396/14,004). Increasing numbers of medications prescribed was associated with decreased knowledge of medications. The strongest predictor of limited knowledge and ability to take medication was dementia (odds ratio > 5.0). Among Ontario seniors living at home, knowledge about medications decreases as the number of medications increases. Therefore, this group may be at high risk of medication errors. Better systems are required to allow healthcare professionals to review with patients, any medications with patients and caregivers, to assist in addressing the decreased knowledge of medications. Such a system would provide the capacity to target those individuals at high risk for a medication error, as well as the medications and drug-drug interactions that seem most likely to be harmful among older adults.


Feasibility of delta-shaped anastomoses in totally laparoscopic distal gastrectomy.
Delta-shaped (DS) anastomosis is a new reconstruction method for totally laparoscopic distal gastrectomy (TLDG) using a linear stapler. We evaluated the feasibility of using this method for TLDG. A retrospective analysis was performed in 114 patients who underwent TLDG with DS anastomosis. Twenty-four patients reconstructed with a Roux-en-Y (RY) anastomosis during the same period were analyzed as control subjects. The patient characteristics of DS and RY anastomoses were slightly different in terms of tumor location and extent of lymph node dissection, since this was not a prospective comparative study. Blood loss, postoperative complication rate and postoperative hospital stay were not different between the two groups. There was only 1 case of anastomotic leakage, and no case of anastomotic stricture after DS anastomosis. The length of the operation using DS anastomosis was significantly shorter than for RY anastomosis. The rates of body weight loss were not significantly different at 1 year after the operation. Although this was a small retrospective analysis, DS anastomosis was feasible, required a shorter operation time, and had no associated complications. This method can therefore be recommended as a standard procedure for TLDG.


Technology-enhanced learning: a role for video animation.
The last 20 years has seen a shift in medical education from printed analogue formats of knowledge transfer to digital knowledge transfer via media platforms and virtual learning environments. Traditional university medical teaching was characterised by lectures and printed textbooks, which to a degree still have an important role to play in knowledge acquisition, but which in isolation do not engage the modern learner, who has become reliant on digital platforms and 'soundbite' learning. Recently, however, traditional methods of teaching and learning have been augmented by, and indeed sometimes replaced by, the alternative learning methods such as: problem-based learning; a greater integration of basic science and clinical considerations; smaller teaching groups; the 'flipped classroom' concept; and various technological tools which promote an interactive learning style. The aim of these new teaching methods is to overcome the well-documented limitations of traditional lectures and printed material in the transfer of knowledge from expert to student, by better engaging the minds of more visual learners and encouraging the use of diverse resources for lifelong learning. In this commentary paper, we share the concept of video animation as an additional educational tool, and one that can help to integrate molecular, cellular and clinical processes that underpin our understanding of biology and pathology in modern education. Importantly, while they can provide focused and attractive formats for 'soundbite' learning, their aim as a tool within the broader educational toolbox is to direct the interested reader towards more traditional formats of learning, which permit a deeper dive into a particular field or concept. In this manner, carefully constructed video animations can serve to provide a broad overview of a particular field or concept and to facilitate deeper learning when desired by the student.


Biochemical lesions in copper-deficient rats caused by secondary iron deficiency. Derangement of protein synthesis and impairment of energy metabolism.
Severe copper deficiency was induced in rats by rearing nursing dams and their offsprings on a semisynthetic diet comprising all the requisite nutrients and trace metals except copper. The copper-deprived rats exhibited growth retardation, severe anaemia, loss of caeruloplasmin, decrease of cytochrome oxidase, accumulation of salt-soluble collagen and a drastic decrease in iron in plasma and liver. Apart from these characteristic signs of deficiency, a marked inhibition of protein synthesis was found to occur both in vivo and in cell-free liver preparations. The curtailed ability to carry out endogenously coded amino acid incorporation into protein contrasted with the unimpaired poly(U)-acid-directed phenylalanine polymerization. This inhibition pattern, as well as the attendant disaggregation of the liver polyribosomes, suggested that the primary biosynthetic lesion was located at the stage of peptide-chain initiation. Concurrently with this alteration there was a pronounced depletion of the hepatic ATP content, associated with a parallel depression of mitochondrial respiration and an enhancement of ATPase activity. Supplementation of the copper-deficient diet with a 2-4-fold excess of iron (relative to the standard diet) prevented growth retardation and anaemia and restored normal energy metabolism, as well as unimpaired protein-synthesizing capacity. The conclusion that these disturbances were primarily determined by the secondary iron deficiency was also borne out by the finding that similar alterations occurred in rats maintained on a copper-sufficient but iron-deficient diet. On the other hand, the iron-fortified diet failed to reverse the other signs of copper deficiency, namely the loss of caeruloplasmin, the diminished rate of cytochrome oxidase and the increase of soluble collagen. The interrelations between the various biochemical lesions induced by deprivation of copper or iron are discussed and the possible role of ATP depletion in determining the derangement of protein synthesis is considered.


Iron and thiol redox signaling in cancer: An exquisite balance to escape ferroptosis.
Epidemiological data indicate a constant worldwide increase in cancer mortality, although the age of onset is increasing. Recent accumulation of genomic data on human cancer via next-generation sequencing confirmed that cancer is a disease of genome alteration. In many cancers, the Nrf2 transcription system is activated via mutations either in Nrf2 or Keap1 ubiquitin ligase, leading to persistent activation of the genes with antioxidative functions. Furthermore, deep sequencing of passenger mutations is clarifying responsible cancer causative agent(s) in each case, including aging, APOBEC activation, smoking and UV. Therefore, it is most likely that oxidative stress is the principal initiating factor in carcinogenesis, with the involvement of two essential molecules for life, iron and oxygen. There is evidence based on epidemiological and animal studies that excess iron is a major risk for carcinogenesis, suggesting the importance of ferroptosis-resistance. Microscopic visualization of catalytic Fe(II) has recently become available. Although catalytic Fe(II) is largely present in lysosomes, proliferating cells harbor catalytic Fe(II) also in the cytosol and mitochondria. Oxidative stress catalyzed by Fe(II) is counteracted by thiol systems at different functional levels. Nitric oxide, carbon monoxide and hydrogen (per)sulfide modulate these reactions. Mitochondria generate not only energy but also heme/iron sulfur cluster cofactors and remain mostly dysfunctional in cancer cells, leading to Warburg effects. Cancer cells are under persistent oxidative stress with a delicate balance between catalytic iron and thiols, thereby escaping ferroptosis. Thus, high-dose L-ascorbate and non-thermal plasma as well as glucose/glutamine deprivation may provide additional benefits as cancer therapies over preexisting therapeutics.


Metabolism of cholesterol and phospholipids in cultured human vascular smooth muscle cells: differences between artery and vein-derived cells and the effect of oxygen partial pressure.
Human smooth muscle (SM) cells derived from vena saphena magna, aorta abdominalis and arteria mamaria were grown in culture under 40 or 145 mmHg oxygen partial pressure (pO2) and their lipid metabolism studied. Esterification of the cellular [3H]cholesterol was higher by 2.5-fold in artery derived than in vein-derived cells and was slightly higher in cultures exposed to 145 mmHg than to 40 mmHg pO2. Cholesterol efflux in the presence of high density lipoprotein (HDL) in the incubation medium was higher in artery-derived than vein-derived cells. Apolipoprotein (apo) AI also supported cholesterol efflux to a higher extent in artery than in vein-derived cells. Cholesterol efflux in the presence of apo AI was accompanied by a decrease of 50% in cellular [3H]cholesteryl ester in both cell types. SM cultures exposed to [3H]choline incorporated about 90% of the radioactivity to phosphatidylcholine (PC) and 10% to sphingomyelin (SPM). During 5 days exposure to [3H]choline, 10 to 15% and 20 to 30% of the newly synthesized PC and SPM, respectively, were released by vein-derived cells into the incubation medium. The relative amount of SPM of the total radioactive phospholipids released by vein-derived cultures was significantly higher in cultures growing under 40 mmHg than 145 mmHg pO2 reaching a value of up to 33% of the radioactive phospholipids in the incubation medium. HDL was shown to serve as an acceptor for phospholipids released by both vein and artery-derived SM cells, while free apo AI supported phospholipid efflux in artery but not in vein-derived SM cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Reflections on Bateson's rule: Solving an old riddle about why extra legs are mirror-symmetric.
William Bateson was an obsessive observer of animal oddities, and at some point in his herculean survey of museum collections leading up to his monumental 1894 monograph (Materials for the study of variation), he noticed a peculiar trend among the preserved specimens (mainly insects) that possessed extra legs: multiple legs that branched from the same socket tended to be mirror images of their adjacent neighbors. He did not know why. These symmetry relationships have come to be known as Bateson's rule, and they have defied a satisfactory explanation for 125 years. In the past few decades, tantalizing clues have emerged from various lines of investigation, and those lines have converged on a possible solution. An attempt is made here to fit all of those clues together to form a coherent picture of the etiology. Two case studies have proven to be pivotal: a fly mutant whose extra legs are caused by patches of dying cells and a frog syndrome whose extra legs are caused by a parasitic flatworm. The conclusion reached is that the extra legs of insects and vertebrates obey Bateson's rule for the same reason, but that reason has nothing to do with the specific molecules in their signaling pathways. Rather, it is an emergent property of the circuitry of the pathways and their polarized alignments along the limb axes. A parade of theoretical models have tried and failed to crack this mystery in the past, and they are reviewed here as part of the narrative.


IFI16 Impacts Metabolic Reprogramming during Human Cytomegalovirus Infection.
Cellular lipid metabolism plays a pivotal role in human cytomegalovirus (HCMV) infection, as increased lipogenesis in HCMV-infected cells favors the envelopment of newly synthesized viral particles. As all cells are equipped with restriction factors (RFs) able to exert a protective effect against invading pathogens, we asked whether a similar defense mechanism would also be in place to preserve the metabolic compartment from HCMV infection. Here, we show that gamma interferon (IFN-γ)-inducible protein 16 (IFI16), an RF able to block HCMV DNA synthesis, can also counteract HCMV-mediated metabolic reprogramming in infected primary human foreskin fibroblasts (HFFs), thereby limiting virion infectivity. Specifically, we find that IFI16 downregulates the transcriptional activation of the glucose transporter 4 (GLUT4) through cooperation with the carbohydrate-response element-binding protein (ChREBP), thereby reducing HCMV-induced transcription of lipogenic enzymes. The resulting decrease in glucose uptake and consumption leads to diminished lipid synthesis, which ultimately curbs the de novo formation of enveloped viral particles in infected HFFs. Consistently, untargeted lipidomic analysis shows enhanced cholesteryl ester levels in IFI16 KO versus wild-type (WT) HFFs. Overall, our data unveil a new role of IFI16 in the regulation of glucose and lipid metabolism upon HCMV replication and uncover new potential targets for the development of novel antiviral therapies. IMPORTANCE Human cytomegalovirus (HCMV) gathers all the substrates and enzymes necessary for the assembly of new virions from its host cell. For instance, HCMV is known to induce cellular metabolism of infected cells to favor virion assembly. Cells are, however, equipped with a first-line defense represented by restriction factors (RFs), which after sensing viral DNA can trigger innate and adaptive responses, thereby blocking HCMV replication. One such RF is IFN-γ-inducible protein 16 (IFI16), which we have shown to downregulate viral replication in human fibroblasts. Thus, we asked whether IFI16 would also play a role in preserving cellular metabolism upon HCMV infection. Our findings highlight an unprecedented role of IFI16 in opposing the metabolic changes elicited by HCMV, thus revealing new promising targets for antiviral therapy.


Visual loss due to optic nerve infarction and central retinal artery occlusion after spine surgery in the prone position: A case report.
Visual loss after spine surgery in the prone position is a serious complication. Several cases of central retinal artery occlusion with ophthalmoplegia after spine surgery have been reported in patients with ophthalmic arteries fed by the internal carotid artery (ICA) in a normal manner. A 74-year-old man developed visual loss after undergoing a spinal decompression and fusion operation in the prone position that lasted approximately 5 hours. We detected an extremely rare case of visual loss due to optic nerve infarction and central retinal artery occlusion through fundoscopic examination, fluorescein angiogram, brain magnetic resonance imaging, and magnetic resonance angiography. The patient's visual loss may have been caused by compromised retrograde collateral circulation of the ophthalmic artery from branches of the external carotid artery in the presence of proximal ICA occlusion after a spinal operation in the prone position. To recover movement of the left extraocular muscles, the patient received intravenous injections of methylprednisolone for 3 days and then oral prednisolone for 6 days. Twenty days after the treatment, the motion of the left extraocular muscles was significantly improved. However, recovery from the left visual loss did not occur until 4 months after the operation. In high-risk patients with retrograde collateral circulation of the ophthalmic artery from the external carotid artery due to proximal ICA occlusion, various measures, including the use of a head fixator to provide a position completely free of direct compression of the head and face, should be considered to decrease the risk of postoperative visual loss.


Review of Physical Layer Security in Molecular Internet of Nano-Things.
Molecular networking has been identified as a key enabling technology for Internet-of-Nano-Things (IoNT): microscopic devices that can monitor, process information, and take action in a wide range of medical applications. As the research matures into prototypes, the cybersecurity challenges of molecular networking are now being researched on at both the cryptographic and physical layer level. Due to the limited computation capabilities of IoNT devices, physical layer security (PLS) is of particular interest. As PLS leverages on channel physics and physical signal attributes, the fact that molecular signals differ significantly from radio frequency signals and propagation means new signal processing methods and hardware is needed. Here, we review new vectors of attack and new methods of PLS, focusing on 3 areas: (1) information theoretical secrecy bounds for molecular communications, (2) key-less steering and decentralized key-based PLS methods, and (3) new methods of achieving encoding and encryption through bio-molecular compounds. The review will also include prototype demonstrations from our own lab that will inform future research and related standardization efforts.


Acute Aortic Valve Rupture From Infective Endocarditis After Transrectal Prostate Biopsy: A Call to Revise the AHA Guidelines for Prevention of Infective Endocarditis.
We describe the case of a 63-year-old man with a known murmur who presented with a 4-month history of intermittent fever and a progressive reduction in energy level after a transrectal prostate biopsy (TRPB). He subsequently presented with acute heart failure secondary to aortic valve cusp rupture caused by endocarditis and underwent urgent aortic valve surgery. The 2008 American College of Cardiology/American Heart Association Guideline update on infective endocarditis has narrowed the use of antibiotic prophylaxis before most accepted dental procedures and continues to recommend against prophylaxis before genitourinary or gastrointestinal procedures. In contrast, the American Urological Association recommends the use of fluoroquinolones for the prevention of transrectal ultrasound-derived infectious complications. Notably, TRPB is associated with a high frequency of bacteremia and bacteriuria. In our case, an antibiotic course before the procedure and a more meticulous medical work-up after febrile illness might have mitigated the patient's catastrophic medical presentation.


Microarray analysis of Foxl2 mediated gene regulation in the mouse ovary derived KK1 granulosa cell line: Over-expression of Foxl2 leads to activation of the gonadotropin releasing hormone receptor gene promoter.
The Foxl2 transcription factor is required for ovarian function during follicular development. The mechanism of Foxl2 regulation of this process has not been elucidated. Our approach to begin to understand Foxl2 function is through the identification of Foxl2 regulated genes in the ovary. Transiently transfected KK1 mouse granulosa cells were used to identify genes that are potentially regulated by Foxl2. KK1 cells were transfected in three groups (mock, activated, and repressed) and twenty-four hours later RNA was isolated and submitted for Affymetrix microarray analysis. Genesifter software was used to carry out analysis of microarray data. One identified target, the gonadotropin releasing hormone receptor (GnRHR) gene, was chosen for further study and validation of Foxl2 responsiveness. Transient transfection analyses were carried out to study the effect of Foxl2 over-expression on GnRHR gene promoter-luciferase fusion activity. Data generated was analyzed with GraphPad Prism software. Microarray analysis identified 996 genes of known function that are potentially regulated by Foxl2 in mouse KK1 granulosa cells. The steroidogenic acute regulatory protein (StAR) gene that has been identified as Foxl2 responsive by others was identified in this study also, thereby supporting the effectiveness of our strategy. The GnRHR gene was chosen for further study because it is known to be expressed in the ovary and the results of previous work has indicated that Foxl2 may regulate GnRHR gene expression. Cellular levels of Foxl2 were increased via transient co-transfection of KK1 cells using a Foxl2 expression vector and a GnRHR promoter-luciferase fusion reporter vector. The results of these analyses indicate that over-expression of Foxl2 resulted in a significant increase in GnRHR promoter activity. Therefore, these transfection data validate the microarray data which suggest that Foxl2 regulates GnRHR and demonstrate that Foxl2 acts as an activator of the GnRHR gene. Potential Foxl2 regulated ovarian genes have been identified through microarray analysis and comparison of these data to other microarray studies. The Foxl2 responsiveness of the GnRHR gene has been validated and provided evidence of Foxl2 transcriptional activation of the GnRHR gene promoter in the mouse ovary derived KK1 granulosa cell line.


From Belly to Brain: Targeting the Ghrelin Receptor in Appetite and Food Intake Regulation.
Ghrelin is the only known peripherally-derived orexigenic hormone, increasing appetite and subsequent food intake. The ghrelinergic system has therefore received considerable attention as a therapeutic target to reduce appetite in obesity as well as to stimulate food intake in conditions of anorexia, malnutrition and cachexia. As the therapeutic potential of targeting this hormone becomes clearer, it is apparent that its pleiotropic actions span both the central nervous system and peripheral organs. Despite a wealth of research, a therapeutic compound specifically targeting the ghrelin system for appetite modulation remains elusive although some promising effects on metabolic function are emerging. This is due to many factors, ranging from the complexity of the ghrelin receptor (Growth Hormone Secretagogue Receptor, GHSR-1a) internalisation and heterodimerization, to biased ligand interactions and compensatory neuroendocrine outputs. Not least is the ubiquitous expression of the GHSR-1a, which makes it impossible to modulate centrallymediated appetite regulation without encroaching on the various peripheral functions attributable to ghrelin. It is becoming clear that ghrelin's central signalling is critical for its effects on appetite, body weight regulation and incentive salience of food. Improving the ability of ghrelin ligands to penetrate the blood brain barrier would enhance central delivery to GHSR-1a expressing brain regions, particularly within the mesolimbic reward circuitry.


Assessing copy number aberrations and copy-neutral loss-of-heterozygosity across the genome as best practice: An evidence-based review from the Cancer Genomics Consortium (CGC) working group for chronic lymphocytic leukemia.
The prognostic role of cytogenetic analysis is well-established in B-cell chronic lymphocytic leukemia (CLL). Approximately 80% of patients have a cytogenetic aberration. Interphase FISH panels have been the gold standard for cytogenetic evaluation, but conventional cytogenetics allows detection of additional abnormalities, including translocations, complex karyotypes and multiple clones. Whole genome copy number assessment, currently performed by chromosomal microarray analysis (CMA), is particularly relevant in CLL for the following reasons: (1) copy number alterations (CNAs) represent key events with biologic and prognostic significance; (2) DNA from fresh samples is generally available; and (3) the tumor burden tends to be relatively high in peripheral blood. CMA also identifies novel copy number variants and copy-neutral loss-of-heterozygosity (CN-LOH), and can refine deletion breakpoints. The Cancer Genomics Consortium (CGC) Working Group for CLL has performed an extensive literature review to describe the evidence-based clinical utility of CMA in CLL. We provide suggestions for the integration of CMA into clinical use and list recurrent copy number alterations, regions of CN-LOH and mutated genes to aid in interpretation.


Diverse retrotransposon families and an AT-rich satellite DNA revealed in giant genomes of Fritillaria lilies.
The genus Fritillaria (Liliaceae) comprises species with extremely large genomes (1C = 30 000-127 000 Mb) and a bicontinental distribution. Most North American species (subgenus Liliorhiza) differ from Eurasian Fritillaria species by their distinct phylogenetic position and increased amounts of heterochromatin. This study examined the contribution of major repetitive elements to the genome obesity found in Fritillaria and identified repeats contributing to the heterochromatin arrays in Liliorhiza species. Two Fritillaria species of similar genome size were selected for detailed analysis, one from each phylogeographical clade: F. affinis (1C = 45·6 pg, North America) and F. imperialis (1C = 43·0 pg, Eurasia). Fosmid libraries were constructed from their genomic DNAs and used for identification, sequence characterization, quantification and chromosome localization of clones containing highly repeated sequences. Repeats corresponding to 6·7 and 4·7 % of the F. affinis and F. imperialis genome, respectively, were identified. Chromoviruses and the Tat lineage of Ty3/gypsy group long terminal repeat retrotransposons were identified as the predominant components of the highly repeated fractions in the F. affinis and F. imperialis genomes, respectively. In addition, a heterogeneous, extremely AT-rich satellite repeat was isolated from F. affinis. The FriSAT1 repeat localized in heterochromatic bands makes up approx. 26 % of the F. affinis genome and substantial genomic fractions in several other Liliorhiza species. However, no evidence of a relationship between heterochromatin content and genome size variation was observed. Also, this study was unable to reveal any predominant repeats which tracked the increasing/decreasing trends of genome size evolution in Fritillaria. Instead, the giant Fritillaria genomes seem to be composed of many diversified families of transposable elements. We hypothesize that the genome obesity may be partly determined by the failure of removal mechanisms to counterbalance effectively the retrotransposon amplification.


The incremental response of random aggregates of identical round particles.
This paper is concerned with a dense, randomly packed, granular material that consists of identical spheres or disks with elastic, frictional interactions, that is first isotropically compressed and subsequently loaded along an arbitrary stress path. An analytical relationship between the overall stress and strain increments is determined for the pre-failure regime. The purpose of the modelling is to understand how this relation depends upon the features of the packing and the particle interactions. From the outset it is recognised that the packing and interactive properties for these materials may vary substantially from grain to grain and the heterogeneity introduced in this manner is fully accounted for. Moment equilibrium equations are solved for each particle and force equilibrium equations are solved for each neighbourhood. Then, the heterogeneity of the aggregate is taken into account by introducing means and fluctuations in the description of the local deformations and the measures of the particles and interactions. The general development is illustrated with an example in two dimensions in which the packing and contact interactions are approximated by angular distributions and the heterogeneity is introduced by variations in these. For an isotropic medium with constant contact stiffnesses the theory provides predictions that compare well with results obtained from numerical simulations.


Assessment of inter-rater agreement on the grading of intravascular bubble signals.
Transcutaneous Doppler ultrasonic bubble detectors are widely used in decompression research. However, interpretation of the complex acoustic signals from the bubble detectors involves a degree of subjectivity, and the comparability of grades assigned by different raters must be assessed. Hypothetical data were used to determine an appropriate method for evaluating the comparability of Doppler raters and to illustrate the limitations of many nonparametric statistics. Two sets of real data were then used to evaluate this procedure, the first from a training exercise carried out by Kisman and Masurel (1978, unpublished) and the second from a test tape that was independently scored by five Defence and Civil Institute of Environmental Medicine Doppler technicians. The results were analyzed by a two-stage approach. First, they were entered into contingency tables and checked for large disagreements, a tendency for one rater to grade higher than the other, and the degree of variability. Second, the results were analyzed with the nonparametric weighted kappa statistic. These studies have led to a practical, efficient method for the evaluation of Doppler raters.


Chlorpyrifos toxicity and detoxifying enzymes activities in three native-aquatic species of macroinvertebrates from an agricultural area.
Non-target species from agricultural areas might be exposed to sublethal pesticide concentrations favoring survival and reproduction of the resistance individuals. The objective of this study was to evaluate chlorpyrifos toxicity and detoxification enzymatic activities on three species (Hyalella curvispina, Heleobia parchappii and Girardia tigrina) from a drain channel with history of insecticide contamination (EF) and the Neuquén river (NR) in Argentina. Chlorpyrifos toxicity on amphipods (H. curvispina) and planarians (G. tigrina) from NR was about six- and two-fold higher than that of their counterparts from EF. Mean carboxylesterases (CarE) activities determined in the three species from NR were significantly different from EF, whereas mean glutathione-S-transferase (GST) activities were no significantly different. Finally, planarians from EF showed significantly higher mean 7-ethoxycoumarine O-deethylase (ECOD) activity than those from NR. Amphipods from both sites displayed similar ECOD activities. The present results suggest that chlorpyrifos resistance in amphipods from EF is not conferred by increased detoxification.


Entropy-driven Adversarial Training For Source-free Medical Image Segmentation.
Although traditional unsupervised domain adaptation (UDA) methods have proven effective in reducing domain gaps, their reliance on source domain data during adaptation often proves unfeasible in real-world applications. For instance, data access in a hospital setting is typically constrained due to patient privacy regulations. To address both the need for privacy protection and the mitigation of domain shifts between source and target domain data, we propose a novel two-step adversarial Source-Free Unsupervised Domain Adaptation (SFUDA) framework in this study. Our approach involves dividing the target domain data into confident and unconfident samples based on prediction entropy, using the Gumbel softmax technique. Confident samples are then treated as source domain data. In order to emulate adversarial training from traditional UDA methods, we employ a min-max loss in the first step, followed by a consistency loss in the second step. Additionally, we introduce a weight to penalize the L2-SP regularizer, which prevents excessive loss of source domain knowledge during optimization. Through extensive experiments on two distinct domain transfer challenges, our proposed SFUDA framework consistently outperforms other SFUDA methods. Remarkably, our approach even achieves competitive results when compared to state-of-the-art UDA methods, which benefit from direct access to source domain data. This demonstrates the potential of our novel SFUDA framework in addressing the limitations of traditional UDA methods while preserving patient privacy in sensitive applications.


Electronic alerts for acute kidney injury.
Acute kidney injury (AKI) is extremely common in hospitalized patients and its presence infers a poorer chance of survival, longer hospital stays and an increased risk of subsequent chronic kidney disease. Recent reports have suggested that standards of care for patients with AKI are often suboptimal and that this contributes to poor outcomes. In an attempt to address delays in diagnosing AKI, there has been increasing interest in e-alert systems for AKI. This review aims to discuss recent developments in e-alert methodologies, as well as examining the evidence to support their effectiveness. Several e-alert systems for AKI have been reported, including more recent descriptions of hospital-wide systems that apply current diagnostic criteria and run sustainably in routine clinical practice. Evidence is accumulating to show their impact in altering physician behaviour and triggering earlier intervention. Emerging data suggest that in combination with other service improvement strategies, this may translate into improved patient outcomes. The current ad-hoc development of e-alert systems needs to be addressed by arriving at consensus around the way in which these systems should apply diagnostic criteria, particularly with respect to selection of baseline creatinine value. Enhancements in IT provision may provide the only durable solution to this, while at the same time facilitating wider uptake. Wider use will allow for further study of the value of e-alerts, including their use in other settings such as primary care.


Gene dosage and complementation analysis of the Shaker locus in Drosophila.
Mutations of the Shaker (Sh) locus alter or eliminate a transient, voltage-sensitive potassium current, the "A" current, in flight muscle of Drosophila. We show that the amplitude of the A current is reduced when the dosage of Sh+ is lower than normal, but that A current amplitude does not increase as extra copies of Sh+ are added. We have also examined 14 Shaker mutants by voltage clamp and by intracellular recording at the larval neuromuscular junction. In 10 of these mutants there is no detectable fast component of the transient outward current. In each of these 10, however, a small, slowly inactivating, outward current is present. The 10 mutations null for the fast component of the transient, outward current are all partially dominant, giving 50-80% of the normal A current in Shnull/Sh+ heterozygotes. Because as little as 5% of the normal A current can be detected, complementation tests are feasible. The 10 null mutations are members of a single complementation group. The remaining 4 mutations have reduced A currents in pupal flight muscle. In all cases, crosses between these leaky mutants and null mutants give progeny with less A current than found in the leaky parental lines, as would be expected if the leaky and null mutations are in the same complementation group. For 1 of the mutations, ShrKO120, the mutant phenotype is much more severe in nerve than in muscle. That part of the Shaker locus required for the production of the A channel lies between the B55 and the V7 translocation breakpoints, in region 16F of the X chromosome.


Prediction of potentially avoidable readmission risk in a division of general internal medicine.
The 30-day post-discharge readmission rate is a quality indicator that may reflect suboptimal care. The computerised algorithm SQLape® can retrospectively identify a potentially avoidable readmission (PARA) with high sensitivity and specificity. We retrospectively analysed the hospital stays of patients readmitted to the Department of Internal Medicine of the CHUV (Centre Hospitalier Universitaire Vaudois) in order to quantify the proportion of PARAs and derive a risk prediction model. All hospitalisations between January 2009 and December 2011 in our division of general internal medicine were analysed. Readmissions within 30 days of discharge were categorised using SQLape®. The impact on PARAs was tested for various clinical and nonclinical factors. The performance of the developed model was compared with the well-validated LACE and HOSPITAL scores. From a total of 11 074 hospital stays, 777 (7%) were followed with PARA within 30 days. By analysing a group of 6729 eligible stays, defined in particular by the patients' returning to their place of residence (home or residential care centre), we identified the following risk factors: ≥1 hospitalisation in the year preceding index admission, Charlson score >1, active cancer, hyponatraemia, length of stay >11 days, prescription of ≥15 different medications during the stay. These variables were used to derive a risk prediction model for PARA with a good discriminatory power (C-statistic 0.70) and calibration (p = 0.69). Patients were then classified as low (16.4%), intermediate (49.4%) or high (34.2%) risk of PARA. The estimated risk of PARA for each category was 3.5%, 8.7% and 19.6%, respectively. The LACE and the HOSPITAL scores were significantly correlated with the PARA risk. The discriminatory power of the LACE (C-statistic 0.61) and the HOSPITAL (C-statistic 0.54) were lower than our model. Our model identifies patients at high risk of 30-day PARA with a good performance. It could be used to target transition of care interventions. Nevertheless, this model should be validated on more data and could be improved with additional parameters. Our results highlight the difficulty to generalise one model in the context of different healthcare systems.


Inhibition of proliferation and survival of diffuse large B-cell lymphoma cells by a small-molecule inhibitor of the ubiquitin-conjugating enzyme Ubc13-Uev1A.
Diffuse large B-cell lymphoma (DLBCL), the most common type of non-Hodgkin lymphoma, remains a partially curable disease. Genetic alterations affecting components of NF-κB signaling pathways occur frequently in DLBCL. Almost all activated B cell-like (ABC) DLBCL, which is the least curable group among the 3 major subtypes of this malignancy, and a substantial fraction of germinal center B cell-like (GCB) DLBCL exhibit constitutive NF-κB pathway activity. It has been demonstrated that ABC-DLBCL cells require such activity for proliferation and survival. Therefore, inhibition of NF-κB activation in DLBCL may provide an efficient and targeted therapy. In screening for small-molecule compounds that may inhibit NF-κB activation in DLBCL cells, we identified a compound, NSC697923, which inhibits the activity of the ubiquitin-conjugating (E2) enzyme Ubc13-Uev1A. NSC697923 impedes the formation of the Ubc13 and ubiquitin thioester conjugate and suppresses constitutive NF-κB activity in ABC-DLBCL cells. Importantly, NSC697923 inhibits the proliferation and survival of ABC-DLBCL cells and GCB-DLBCL cells, suggesting the Ubc13-Uev1A may be crucial for DLBCL growth. Consistently, knockdown of Ubc13 expression also inhibited DLBCL cell survival. The results of the present study indicate that Ubc13-Uev1A may represent a potential therapeutic target in DLBCL. In addition, compound NSC697923 may be exploited for the development of DLBCL therapeutic agents.


Alternative Medicine in Diabetes - Role of Angiogenesis, Oxidative Stress, and Chronic Inflammation.
Diabetes is a chronic metabolic disorder that is characterized by hyperglycemia due to lack of or resistance to insulin. Patients with diabetes are frequently afflicted with ischemic vascular disease and impaired wound healing. Type 2 diabetes is known to accelerate atherosclerotic processes, endothelial cell dysfunction, glycosylation of extracellular matrix proteins, and vascular denervation. Herbal medicines and naturally occurring substances may positively affect diabetes management, and could thus be utilized as cost-effective means of supporting treatment in developing countries. Natural treatments have been used in these countries for a long time to treat diabetes. The present review analyses the features of aberrant angiogenesis, abnormalities in growth factors, oxidative stress, and metabolic derangements relevant to diabetes, and how herbal substances and their active chemical constituents may counteract these events. Evidence for possible biochemical effectiveness and limitations of herbal medicines are given, as well as details regarding the role of cytokines and nitric oxide.


Procoagulant and anticoagulant factors in childhood hypothyroidism.
The aim of this study was to investigate the effects of thyroid hormone deficiencies in childhood on the elements of coagulation proteins. Consecutive 54 children with hypothyroidism and 55 healthy controls aged 1 month-16 years were enrolled. One year after Na-L-thyroxine treatment, the study parameters were reevaluated. Thyroid function tests, procoagulant and anticoagulant proteins were performed for children with hypothyroidism and healthy controls. Significant decreased results were found in children with hypothyroidism in terms of fibrinogen, TT, and anticoagulant proteins including AT, PC, PS, and fPS. Significant increases were found with respect to APTT, fibrinogen, and TT. In the evaluation of posttreatment changes a statistically significant increase was found in vWF, FVIII, AT, PC, PS, and fPS. A positive correlation was found between fT4 and vWF, FVIII, PC, and PS. We would like to emphasize that the coagulation system especially vWF and FVIII, and particularly the anticoagulant system, should be monitored closely in patients followed up for hypothyroidism. Thyroid hormones should be examined and, if necessary, hormone replacement therapy should be administered in patients followed up for a predisposition to coagulation. Additionally, further studies with larger series are needed to investigate the effects of hypothyroidism on the coagulation system.


A Corticostriatal Balancing Act Supports Skill Learning.
In this issue of Neuron, Kupferschmidt et al. (2017) reveal the shifting dynamics of functionally defined corticostriatal pathways during skill learning in mice using fiber photometry. They show different time courses in plasticity of associative and sensorimotor circuits across learning that involve changes at both the synaptic and cortical level.


Implementation of an intersectoral program to eliminate human and canine rabies: the Bohol Rabies Prevention and Elimination Project.
The province of Bohol, located in the Visayas islands region in the Philippines has a human population of 1.13 million and was the 4th highest region for human rabies deaths in the country, averaging 10 per year, prior to the initiation of the Bohol Rabies Prevention and Elimination Project (BRPEP). The BRPEP was initiated in 2007 with the goal of building a sustainable program that would prevent human rabies by eliminating rabies at its source, in dogs, by 2010. This goal was in line with the Philippine National Rabies Program whose objective is to eliminate rabies by 2020. The intersectoral BRPEP was launched in 2007 and integrated the expertise and resources from the sectors of agriculture, public health and safety, education, environment, legal affairs, interior and local government. The program included: increasing local community involvement; implementing dog population control; conducting mass dog vaccination; improving dog bite management; instituting veterinary quarantine; and improving diagnostic capability, surveillance and monitoring. Funding was secured from the national government, provincial, municipal and village units, dog owners, NGOs, the regional office of the WHO, the UBS Optimus Foundation, and the Global Alliance for Rabies Control. The BRPEP was managed by the Bohol Rabies Prevention and Eradication Council (BRPEC) under the jurisdiction of the Governor of Bohol. Parallel organizations were created at the municipal level and village level. Community volunteers facilitated the institution of the program. Dog population surveys were conducted to plan for sufficient resources to vaccinate the required 70% of the dogs living in the province. Two island-wide mass vaccination campaigns were conducted followed by "catch up" vaccination campaigns. Registration of dogs was implemented including a small fee that was rolled back into the program to maintain sustainability. Children were educated by introducing rabies prevention modules into all elementary schools in Bohol. Existing public health legislation at the national, provincial, and municipal level strengthened the enforcement of activities. A Knowledge, Attitude and Practices (KAP) survey was conducted in 2009 to evaluate the educational knowledge of the population. Increased surveillance was instituted to ensure that dogs traveling into and out of the province were vaccinated against rabies. Human and animal cases of rabies were reported to provincial and national authorities. Within the first 18 months of the BRPEP, human rabies deaths had decreased annually from 0.77 to 0.37 to zero per 100,000 population from 2007-2009. Between October 2008 and November 2010 no human and animal cases were detected. Increased surveillance on the island detected one suspected human rabies case in November 2010 and one confirmed case of canine rabies in April 2011. Two mass vaccination campaigns conducted in 2007 and 2008 successfully registered and vaccinated 44% and 70% of the dogs on the island. The additional surveillance activities enabled a mobilization of mop up vaccination activities in the region where the human and canine case was located. Due to the increased effective and continuous surveillance activities, rabies was stopped before it could spread to other areas on the island. The program costs totaled USD 450,000. Registration fees collected to maintain the program amounted to USD 105,740 and were re-allocated back into the community to sustain the program.


CGRP and Shh Mediate the Dental Pulp Cell Response to Neuron Stimulation.
Dental pain is a persistent, detrimental public health issue that requires a better understanding of the mechanisms of tooth pain and inflammation in order to develop more effective treatments. Calcitonin gene-related peptide (CGRP) and dental pulp cells are promising candidates for mediating tooth pain and generating reparative dental tissues, respectively, but their behavior in the context of pulpitis remains elusive. The mouse incisor requires Sonic hedgehog (Shh) secreted from sensory nerves to continuously regenerate. However, it is unknown whether sensory nerves also regulate the comparatively nonregenerative mouse molar through CGRP and Shh. This is an important knowledge gap to fill since mouse incisors differ biologically from human teeth, while mouse and human molars are similar. In this work, we identified that molar pulp cells express CGRP receptor and Gli1, a Hedgehog (Hh) signaling protein found to label a dental stem cell population in the mouse incisor. We also observed in a mouse molar injury model that Hh signaling was activated and Shh expression was upregulated in vivo. We then determined in vitro that Shh and CGRP regulate differentiation of primary mouse molar and incisor pulp cells and a human dental pulp stem cell line. Furthermore, conditioned media from stimulated sensory neurons induced Hh signaling activation and inflammatory gene expression in primary molar pulp cells, which was abolished by inhibition of either Shh or CGRP. Our results suggest that CGRP and Shh signaling may promote an inflammatory response after injury in the molar and that activated sensory nerves secrete CGRP and Shh to regulate molar pulp cell expansion and differentiation into odontoblast-like cells for dentin repair. Thus, CGRP/Shh signaling should be considered for new strategies that seek to manage pain or dentin regeneration in the molar.


The 9p21 region in bladder cancer cell lines: large homozygous deletion inactivate the CDKN2, CDKN2B and MTAP genes.
Homozygous and hemizygous deletions of 9p21 are the earliest and most common genetic alteration in bladder cancer. The identification of two cell cycle regulators, CDKN2 and CDKN2B, that map to the common region of deletion has prompted the hypothesis that they are critical tumor suppressor genes in this malignancy. However, controversy as to whether these genes are the only or even the most important target in bladder cancer oncogenesis remains. To more clearly determine the effect of these 9p21 alterations, we mapped the homozygous deletions and performed a detailed mutational and expression analysis for CDKN2, CDKN2B and a closely linked gene, methylthioadenoside phosphorylase (MTAP), in 16 established bladder cancer cell lines. Nine of the 16 lines exhibit large (30 to > 2000 kb) homozygous deletions on 9p21. All deletions include at least one exon of CDKN2, eight of nine include CDKN2B, and six of nine include MTAP. MTAP function correlates with the genomic deletions. SSCP and sequence analysis does not reveal any inactivating point mutations of CDKN2 or of CDKN2B in any of the cell lines without homozygous deletions, and all express the CDKN2 and the CDKN2B mRNA as well as the encoded p16 protein. The p16 protein levels vary widely and are correlated with absent pRb expression. We conclude that the 9p21 deletions in bladder cancer usually inactivate the CDKN2. CDKN2B, and MTAP genes but that CDKN2 is the most common target. Other mechanisms for inactivating this gene in bladder cancer appear to be uncommon.


Low-spin ferric forms of cytochrome a3 in mixed-ligand and partially reduced cyanide-bound derivatives of cytochrome c oxidase.
Optical-absorption-, e.p.r.- and m.c.d. (magnetic-circular-dichroism)-spectroscopic measurements were made on liganded derivatives of oxidized and partially reduced cytochrome c oxidase. When NO was added to oxidized cyanide-bound cytochrome c oxidase, no changes occurred in the optical-absorption difference spectrum. In contrast, NO induced reduction of cytochrome a3 and formation of the nitrosylferrohaem species when the oxidized resting enzyme was the starting material. E.p.r. spectroscopy of the NO-treated oxidized cyanide-bound enzyme revealed the presence of a low-spin haem signal at g = 3.40, whereas the g = 3.02 and g = 2.0 signals of the oxidized enzyme remained unchanged. Both haem groups in this species are e.p.r.-detectable simultaneously. Examination of an identical sample by m.c.d. spectroscopy in the near-i.r. region identified two distinct low-spin species at 1565 and 1785 nm. Irradiation with white light of the NO-treated cyanide-bound sample at 10K resulted in the disappearance of the g = 3.40 e.p.r. signal and the m.c.d. signal at 1785 nm, whereas a band at 1950nm increased in intensity. When the photolysed sample was warmed to 50K and held in the dark for 15 min, the original spectrum returned. Magnetization studies of the 1785nm m.c.d. band support the assignment of this signal to the same metal centre that gives rise to the g = 3.40 e.p.r. signal. The effect of NO on the oxidized cyanide-bound enzyme was compared with that obtained when the oxidized cyanide-bound species was taken to the partially reduced state. Cytochrome a3 is e.p.r.-detectable with a g-value of 3.58 [Johnson, Eglinton, Gooding, Greenwood & Thomson (1981) Biochem. J. 193, 699-708]. Its near-i.r. m.c.d. spectrum shifts from 1950nm in the oxidized cyanide-bound enzyme to 1545nm on addition of reductant. A scheme is advanced for the structure of the cytochrome a3-CuB site that allows for cyanide binding to Fea3 and NO binding to CuB. Cyanide is the bridging ligand in the ferromagnetically coupled cytochrome a3-CuB pair of oxidized cyanide-bound cytochrome c oxidase. The bridged structure and the magnetic interaction are broken when the enzyme is partially reduced. However, when NO binds to CuB the cyanide bridge remains intact, but now the odd spins of NO and CuB are magnetically coupled.