Lack of effect of methylprednisolone on lysosomal and microsomal enzymes after two hours of well-defined canine myocardial ischemia.
Myocardial ischemia was produced for 2 hours by coronary ligation in 11 dogs pretreated with methylprednisolone (MP, 30 mg/kg). Myocardial blood flow (MBF) was measured with microspheres (15 micrometer) in each tissue sample used for enzymatic analysis. Homogenates of these tissue samples were separated by ultracentrifugation into lysosome-rich and microsomal fractions and were analyzed for N-acetyl-beta-glusosaminidase (NAGA), beta-glucuronidase (beta-gluc), rotenone-insensitive-NADH-cytochrome c reductase (RINCR), and cytochrome oxidase. The enzymatic data from centrifugal fractions were grouped according to MBF values for statistical analysis of inter-group effects of ischemia. Significant losses (P less than 0.001) of NAGA and beta-gluc were seen in all MP-treated lysosome-rich particulate fractions that were isolated from zones demonstrating MBF values less than 25% of control (L-ischemia). Similar significant losses (P less than 0.001) of RINCR were seen in microsomal fractions from L-ischemia zones. Samples with MBF values greater than 25% but less than 75% of control (M-ischemia) also demonstrated significant decreases of lysosomal and microsomal enzymatic activity in specific fractions. When the data of the above MP-treated group were compared with the untreated control group, no significant intergroup effects of treatment with MP were observed. In addition, enzymatic data (NAGA, RINCR) were normalized prior to performing linear regression analyses; percent loss of particulate enzymatic activity was plotted against percent decrease in MBF. The effects of 2 hours of ischemia on the above biochemical parameters were comparable between untreated and MP-treated groups. Finally, when myocardial samples were grouped according to similar levels of MBF, statistical analysis using the general linear models procedure revealed no beneficial effect of MP treatment on changes in lysosomal hydrolases, microsomal RINCR, or latency of lysosomes.


Disentangling neural representations of value and salience in the human brain.
A large body of evidence has implicated the posterior parietal and orbitofrontal cortex in the processing of value. However, value correlates perfectly with salience when appetitive stimuli are investigated in isolation. Accordingly, considerable uncertainty has remained about the precise nature of the previously identified signals. In particular, recent evidence suggests that neurons in the primate parietal cortex signal salience instead of value. To investigate neural signatures of value and salience, here we apply multivariate (pattern-based) analyses to human functional MRI data acquired during a noninstrumental outcome-prediction task involving appetitive and aversive outcomes. Reaction time data indicated additive and independent effects of value and salience. Critically, we show that multivoxel ensemble activity in the posterior parietal cortex encodes predicted value and salience in superior and inferior compartments, respectively. These findings reinforce the earlier reports of parietal value signals and reconcile them with the recent salience report. Moreover, we find that multivoxel patterns in the orbitofrontal cortex correlate with value. Importantly, the patterns coding for the predicted value of appetitive and aversive outcomes are similar, indicating a common neural scale for appetite and aversive values in the orbitofrontal cortex. Thus orbitofrontal activity patterns satisfy a basic requirement for a neural value signal.


Improving self-help e-therapy for depression and anxiety among sexual minorities: an analysis of focus groups with lesbians and gay men.
E-therapies for depression and anxiety rarely account for lesbian and gay users. This is despite lesbians and gay men being at heightened risk of mood disorders and likely to benefit from having access to tailored self-help resources. We sought to determine how e-therapies for depression and anxiety could be improved to address the therapeutic needs of lesbians and gay men. We conducted eight focus groups with lesbians and gay men aged 18 years and older. Focus groups were presented with key modules from the popular e-therapy "MoodGYM". They were asked to evaluate the inclusiveness and relevance of these modules for lesbians and gay men and to think about ways that e-therapies in general could be modified. The focus groups were analyzed qualitatively using a thematic analysis approach to identify major themes. The focus groups indicated that some but not all aspects of MoodGYM were suitable, and suggested ways of improving e-therapies for lesbian and gay users. Suggestions included avoiding language or examples that assumed or implied users were heterosexual, improving inclusiveness by representing non-heterosexual relationships, providing referrals to specialized support services and addressing stigma-related stress, such as "coming out" and experiences of discrimination and harassment. Focus group participants suggested that dedicated e-therapies for lesbians and gay men should be developed or general e-therapies be made more inclusive by using adaptive logic to deliver content appropriate for a user's sexual identity. Findings from this study offer in-depth guidance for developing e-therapies that more effectively address mental health problems among lesbians and gay men.


Differences in Balance Function Between Cancer Survivors and Healthy Subjects: A Pilot Study.
Older adults who have survived cancer experience significantly more falls compared with healthy adults. Adult cancer survivors may also have a lower balance function than healthy adults. We examined muscle strength and balance function among 19 cancer survivors and 14 healthy subjects. The mean age of the cancer survivors was 51.5 ± 11.2 years; 6 men and 13 women. Cancer diagnoses included breast cancer, retroperitoneal sarcoma, acute leukemia, lung cancer, colorectal cancer, thyroid cancer, Ewing's sarcoma, and tongue cancer. The mean age of healthy subjects was 47.4 ± 14 years; 3 men, 11 women. Muscle strength was assessed using hand grip and knee extensor strength tests. Balance function was evaluated using the Timed Up and Go (TUG) test, and body sway was tested using a force platform. No significant differences were found with respect to right and left grip strength or right and left knee extension strength between the 2 groups. A significantly higher TUG time was observed in cancer survivors than in healthy subjects ( P < .05). With eyes open, the area of the center of pressure was significantly larger in cancer survivors than in healthy subjects ( P < .05). Similarly, the length per area was significantly lower both with eyes open and closed for cancer survivors than for healthy subjects ( P < .05). TUG was significantly correlated with muscle strength in both groups ( P < .05). However, no body sway parameters were related to muscle strength in either group. Cancer survivors had lower balance function that might not have been related to muscle strength. Cancer survivors should be evaluated for balance function as there is a potential for impairment. The findings of this study will be relevant for planning the prevention of falls for cancer survivors.


Chemokine CCL4 Induces Vascular Endothelial Growth Factor C Expression and Lymphangiogenesis by miR-195-3p in Oral Squamous Cell Carcinoma.
The inflammatory chemokine (C-C motif) ligand 4 (CCL4) plays an important role in the pathogenesis and progression of cancer. In particular, higher serum CCL4 levels in patients with oral squamous cell carcinoma (OSCC) are associated with a more advanced stage of disease. OSCC accounts for approximately 95% of oral cancer in Taiwan and has a poor prognosis, due to aggressive local invasion and metastasis, leading to recurrence. OSCC spreads preferentially through lymphatic vessels and has the propensity to metastasize to the cervical lymph nodes even in the early stage of disease. Vascular endothelial growth factor C (VEGF-C) is an essential regulator of lymphangiogenesis. In particular, VEGF-C is specific to lymphatic vessel development, and VEGF-C expression levels have been found to directly correlate with lymph node metastasis in OSCC. However, it is unclear as to whether CCL4 correlates with VEGF-C expression and lymphangiogenesis in OSCC. We found that CCL4 increased VEGF-C expression and promoted lymphangiogenesis in oral cancer cells in vitro and in vivo. miR-195-3p mimic reversed CCL4-enhanced VEGF-C expression. CCL4 stimulation of oral cancer cells augmented JAK2 and STAT3 phosphorylation. Thus, CCL4 may be a new molecular therapeutic target for inhibition of lymphangiogenesis and metastasis in OSCC.


How early is too early? Challenges in ART initiation and engaging in HIV care under Treat All in Rwanda-A qualitative study.
HIV treatment guidelines recommend that all people living with HIV (PLWH) initiate antiretroviral therapy (ART) as soon as possible after diagnosis (Treat All). As Treat All is more widely implemented, an increasing proportion of PLWH are likely to initiate ART when they are asymptomatic, and they may view the relative benefits and risks of ART differently than those initiating at more advanced disease stages. To date, patient perspectives of initiating care under Treat All in sub-Saharan Africa have not been well described. From September 2018 to March 2019, we conducted individual, semi-structured, qualitative interviews with 37 patients receiving HIV care in two health centers in Kigali, Rwanda. Data were analyzed using a mixed deductive and inductive thematic analysis approach to describe perceived barriers to, facilitators of and acceptability of initiating and adhering to ART rapidly under Treat All. Of 37 participants, 27 were women and the median age was 31 years. Participants described feeling traumatized and overwhelmed by their HIV diagnosis, resulting in difficulty accepting their HIV status. Most were prescribed ART soon after diagnosis, yet fear of lifelong medication and severe side effects in the immediate period after initiating ART led to challenges adhering to therapy. Moreover, because many PLWH initiated ART while healthy, taking medications and attending appointments were visible signals of HIV status and highly stigmatizing. Nonetheless, many participants expressed enthusiasm for Treat All as a program that improved health as well as health equity. For newly-diagnosed PLWH in Rwanda, initiating ART rapidly under Treat All presents logistical and emotional challenges despite the perceived benefits. Our findings suggest that optimizing early engagement in HIV care under Treat All requires early and ongoing intervention to reduce trauma and stigma, and promote both individual and community benefits of ART.


Third-harmonic generation for the study of Caenorhabditis elegans embryogenesis.
Live microscopy techniques (i.e., differential interference contrast, confocal microscopy, etc.) have enabled the understanding of the mechanisms involved in cells and tissue formation. In long-term studies, special care must be taken in order to avoid sample damage, restricting the applicability of the different microscopy techniques. We demonstrate the potential of using third-harmonic generation (THG) microscopy for morphogenesis/embryogenesis studies in living Caenorhabditis elegans (C. elegans). Moreover, we show that the THG signal is obtained in all the embryo development stages, showing different tissue/structure information. For this research, we employ a 1550-nm femtosecond fiber laser and demonstrate that the expected water absorption at this wavelength does not severely compromise sample viability. Additionally, this has the important advantage that the THG signal is emitted at visible wavelengths (516 nm). Therefore, standard collection optics and detectors operating near maximum efficiency enable an optimal signal reconstruction. All this, to the best of our knowledge, demonstrates for the first time the noninvasiveness and strong potential of this particular wavelength to be used for high-resolution four-dimensional imaging of embryogenesis using unstained C. elegans in vivo samples.


Targeted Next Generation Sequencing Identifies Markers of Response to PD-1 Blockade.
Therapeutic antibodies blocking programmed death-1 and its ligand (PD-1/PD-L1) induce durable responses in a substantial fraction of melanoma patients. We sought to determine whether the number and/or type of mutations identified using a next-generation sequencing (NGS) panel available in the clinic was correlated with response to anti-PD-1 in melanoma. Using archival melanoma samples from anti-PD-1/PD-L1-treated patients, we performed hybrid capture-based NGS on 236-315 genes and T-cell receptor (TCR) sequencing on initial and validation cohorts from two centers. Patients who responded to anti-PD-1/PD-L1 had higher mutational loads in an initial cohort (median, 45.6 vs. 3.9 mutations/MB; P = 0.003) and a validation cohort (37.1 vs. 12.8 mutations/MB; P = 0.002) compared with nonresponders. Response rate, progression-free survival, and overall survival were superior in the high, compared with intermediate and low, mutation load groups. Melanomas with NF1 mutations harbored high mutational loads (median, 62.7 mutations/MB) and high response rates (74%), whereas BRAF/NRAS/NF1 wild-type melanomas had a lower mutational load. In these archival samples, TCR clonality did not predict response. Mutation numbers in the 315 genes in the NGS platform strongly correlated with those detected by whole-exome sequencing in The Cancer Genome Atlas samples, but was not associated with survival. In conclusion, mutational load, as determined by an NGS platform available in the clinic, effectively stratified patients by likelihood of response. This approach may provide a clinically feasible predictor of response to anti-PD-1/PD-L1. Cancer Immunol Res; 4(11); 959-67. ©2016 AACR.


Genome divergence in two Prochlorococcus ecotypes reflects oceanic niche differentiation.
The marine unicellular cyanobacterium Prochlorococcus is the smallest-known oxygen-evolving autotroph. It numerically dominates the phytoplankton in the tropical and subtropical oceans, and is responsible for a significant fraction of global photosynthesis. Here we compare the genomes of two Prochlorococcus strains that span the largest evolutionary distance within the Prochlorococcus lineage and that have different minimum, maximum and optimal light intensities for growth. The high-light-adapted ecotype has the smallest genome (1,657,990 base pairs, 1,716 genes) of any known oxygenic phototroph, whereas the genome of its low-light-adapted counterpart is significantly larger, at 2,410,873 base pairs (2,275 genes). The comparative architectures of these two strains reveal dynamic genomes that are constantly changing in response to myriad selection pressures. Although the two strains have 1,350 genes in common, a significant number are not shared, and these have been differentially retained from the common ancestor, or acquired through duplication or lateral transfer. Some of these genes have obvious roles in determining the relative fitness of the ecotypes in response to key environmental variables, and hence in regulating their distribution and abundance in the oceans.


Investigating selected host and parasite factors potentially impacting upon seasonal malaria chemoprevention in Bama, Burkina Faso.
Since 2014, seasonal malaria chemoprevention (SMC) with amodiaquine-sulfadoxine-pyrimethamine (AQ-SP) has been implemented on a large scale during the high malaria transmission season in Burkina Faso. This paper reports the prevalence of microscopic and submicroscopic malaria infection at the outset and after the first round of SMC in children under 5 years old in Bama, Burkina Faso, as well as host and parasite factors involved in mediating the efficacy and tolerability of SMC. Two sequential cross-sectional surveys were conducted in late July and August 2017 during the first month of SMC in a rural area in southwest Burkina Faso. Blood smears and dried blood spots were collected from 106 to 93 children under five, respectively, at the start of SMC and again 3 weeks later. Malaria infection was detected by microscopy and by PCR from dried blood spots. For all children, day 7 plasma concentrations of desethylamodiaquine (DEAQ) were measured and CYP2C8 genetic variants influencing AQ metabolism were genotyped. Samples were additionally genotyped for pfcrt K76T and pfmdr1 N86Y, molecular markers associated with reduced amodiaquine susceptibility. 2.8% (3/106) of children were positive for Plasmodium falciparum infection by microscopy and 13.2% (14/106) by nested PCR within 2 days of SMC administration. Three weeks after SMC administration, in the same households, 4.3% (4/93) of samples were positive by microscopy and 14.0% (13/93) by PCR (p = 0.0007). CYP2C8*2, associated with impaired amodiaquine metabolism, was common with an allelic frequency of 17.1% (95% CI 10.0-24.2). Day 7 concentration of DEAQ ranged from 0.48 to 362.80 ng/mL with a median concentration of 56.34 ng/mL. Pfmdr1 N86 predominated at both time points, whilst a non-significant trend towards a higher prevalence of pfcrt 76T was seen at week 3. This study showed a moderate prevalence of low-level malaria parasitaemia in children 3 weeks following SMC during the first month of administration. Day 7 concentrations of the active DEAQ metabolite varied widely, likely reflecting variability in adherence and possibly metabolism. These findings highlight factors that may contribute to the effectiveness of SMC in children in a high transmission setting.


Rectus abdominis muscle resection for abdominal wall recurrence of mucinous adenocarcinoma or peritoneal mesothelioma.
Diagnostic technologies which penetrate the abdominal wall in an attempt to definitively diagnose an intraabdominal malignancy by biopsy can contaminate the abdominal wall by cancerous cells. With follow-up these entrapped cancer cells may progress as an abdominal wall recurrence of the disease process. Frequently, laparoscopy is the definitive diagnostic study which results in the abdominal wall cancer progression. We examined recurrences within the abdominal wall and attempted to establish a surgical approach to this problem which would maximize a functional result and minimize the incidence of disease persistence within the abdominal wall. Eighteen patients with abdominal wall recurrence were studied. Laparoscopy port sites resulted in the abdominal wall disease in eight patients, in four the recurrence was at a previous ostomy site, in three it was in a Pfannenstiel incision and in three it was in a McBurney incision site. All of these patients were treated by total resection of the rectus abdominis muscle. This resulted in a complete removal of visible disease that was dissecting along the fibers of the rectus abdominis muscle. No patients required reoperation for abdominal wall hernia and mesh repair was not used in any of these patients. Disease control within the abdominal wall has been excellent.


The Relationship Between Alexithymia and Emotional Awareness: A Meta-Analytic Review of the Correlation Between TAS-20 and LEAS.
Background: Alexithymia and emotional awareness may be considered overlapping constructs and both have been shown to be related to psychological and emotional well-being. However, it is not clear how the constructs relate to each other empirically or if they may overlap more or less in different populations. The aim of this review was therefore to conduct a meta-analysis of correlations between the most commonly used measures of alexithymia (i.e., the self-report instrument Toronto Alexithymia Scale; TAS-20) and emotional awareness (i.e., the observer-rated instrument Level of Emotional Awareness Scale; LEAS) and to explore potential moderators of their relationship. Methods: Electronic databases were searched for studies published until the end of February 2018. Study samples were coded as medical conditions, psychiatric disorders and/or healthy controls and sample mean age and gender distribution were extracted. Correlations between the TAS-20 and the LEAS were subjected to a random effect of meta-analysis and moderators were explored in subgroup analyses and meta-regressions. Publication bias was considered. Results: 21 studies reporting on 28 independent samples on correlation analysis were included, encompassing a total of 2857 subjects (57% women). The aggregated correlation between TAS-20 and LEAS was r = -0.122 (95% CI [-0.180, -0.064]; Z = -4.092; p < 0.001), indicating a significant, but weak, negative relationship between the measures. Heterogeneity was moderate, but we found no indication of significant differences between patients with medical conditions, psychiatric disorders or healthy controls, nor that mean age or percentage of female subjects moderated the relationship. The overall estimate became somewhat weaker after adjusting for possible publication bias. Conclusions: Our results indicate that TAS-20 and LEAS measure different aspects of emotional functioning. The small overlap suggests that alexithymia and emotional awareness are distinct constructs of emotional well-being. Clinicians need to assess both aspects when considering treatment options for individual patients. Moreover, from the clinical standpoint, an easy reliable and valid way of measuring emotional awareness is still needed. More research should be focus on the differences between alexithymia and emotional awareness in specific conditions, but also how to integrate self-report instrument and observed based measures in a clinical situation.


Single and multiple human papillomavirus infections in cervical abnormalities in Portuguese women.
Persistent infection with high-risk (HR) human papillomavirus (HPV) types is necessary for cervical cancer development. However, little is known about the influence of multiple HPV infections on cervical lesion risk. The aim of this study was to evaluate the frequency of single and multiple HPV infections in Portuguese women, and to assess the frequency of multiple infections in cervical intraepithelial neoplasia (CIN). HPV prevalence, type-specific prevalence and extent of multiple infections were assessed in 1057 cervical samples. The Clinical Array HPV assay was used to detect 35 HPV types. According to histological diagnosis, 425 samples were normal, 375 were CIN1, and 257 were CIN2+. HPV status was studied in relation to age and lesion severity. The prevalence of HPV infection was 52.7%; 25.4%, 67.2% and 76.7% were positive for any HPV type in the normal, CIN1 and CIN2+ cases, respectively. Among HPV-positive cases, 32.0% were associated with multiple infections. Among multiple infections, 96.1% harboured HR HPV types and 38.2% HR-low risk (LR) HPV types. Overall, 33 different HPV types (18 HR and 15 LR) were detected. HR HPV types (44.1%) were significantly more prevalent than LR HPV types (8.6%). The most frequent genotype was HPV 16 (25.5%), followed by HPV 31, 53, 66, 58, and 51. Multiple infections showed a significant increase (p 0.005) according to severity of neoplasia, particularly for HR-HR HPV infections (p 0.003). No association between age and multiple HPV infections was observed (p 0.812). However, multiple HR HPV infections were more frequent in women under 30 years of age (35.3%).


A comparative modeling study on non-climatic and climatic risk assessment on Asian Tiger Mosquito (Aedes albopictus).
Aedes albopictus, the Asian Tiger Mosquito, vector of Chikungunya, Dengue Fever and Zika viruses, has proven its hardy adaptability in expansion from its natural Asian, forest edge, tree hole habitat on the back of international trade transportation, re-establishing in temperate urban surrounds, in a range of water receptacles and semi-enclosures of organic matter. Conventional aerial spray mosquito vector controls focus on wetland and stagnant water expanses, proven to miss the protected hollows and crevices favoured by Ae. albopictus. New control or eradication strategies are thus essential, particular in light of potential expansions in the southeastern and eastern USA. Successful regional vector control strategies require risk level analysis. Should strategies prioritize regions with non-climatic or climatic suitability parameters for Ae. albopictus? Our study used current Ae. albopictus distribution data to develop two independent models: (i) regions with suitable non-climatic factors, and (ii) regions with suitable climate for Ae. albopictus in southeastern USA. Non-climatic model processing used Evidential Belief Function (EBF), together with six geographical conditioning factors (raster data layers), to establish the probability index. Validation of the analysis results was estimated with area under the curve (AUC) using Ae. albopictus presence data. Climatic modeling was based on two General Circulation Models (GCMs), Miroc3.2 and CSIRO-MK30 running the RCP 8.5 scenario in MaxEnt software. EBF non-climatic model results achieved a 0.70 prediction rate and 0.73 success rate, confirming suitability of the study site regions for Ae. albopictus establishment. The climatic model results showed the best-fit model comprised Coldest Quarter Mean Temp, Precipitation of Wettest Quarter and Driest Quarter Precipitation factors with mean AUC value of 0.86. Both GCMs showed that the whole study site is highly suitable and will remain suitable climatically, according to the prediction for 2055, for Ae. albopictus expansion.


Vezf1 protein binding sites genome-wide are associated with pausing of elongating RNA polymerase II.
The protein Vezf1 plays multiple roles important for embryonic development. In Vezf1(-/-) mouse embryonic stem (mES) cells, our earlier data showed widespread changes in gene-expression profiles, including decreased expression of the full-length active isoform of Dnmt3b methyltransferase and concomitant genome-wide reduction in DNA methylation. Here we show that in HeLaS3 cells there is a strong genome-wide correlation between Vezf1 binding and peaks of elongating Ser2-P RNA polymerase (Pol) ll, reflecting Vezf1-dependent slowing of elongation. In WT mES cells, the elongating form of RNA pol II accumulates near Vezf1 binding sites within the dnmt3b gene and at several other Vezf1 sites, and this accumulation is significantly reduced at these sites in Vezf1(-/-) mES cells. Depending upon genomic location, Vezf1-mediated Pol II pausing can have different regulatory roles in transcription and splicing. We find examples of genes in which Vezf1 binding sites are located near cassette exons, and in which loss of Vezf1 leads to a change in the relative abundance of alternatively spliced messages. We further show that Vezf1 interacts with Mrg15/Mrgbp, a protein that recognizes H3K36 trimethylation, consistent with the role of histone modifications at alternatively spliced sites.


Secondary school teachers' experiences related to learner teenage pregnancies and unexpected deliveries at school.
The incidence of learner teenage pregnancies can be reduced, provided that the major stakeholders, which include the Department of Education and the Department of Health, combine to address this issue. Despite the implementation of Life Orientation as a school subject, which focuses on sexual behaviour, health, decision-making, pregnancy risk, sexually transmitted infections and HIV and AIDS, the prevalence of learner teenage pregnancies at secondary schools remains alarmingly high. The purpose of this study was to explore and describe teachers' experiences of learner teenage pregnancies in secondary schools in a province in South Africa. Secondary schools in KwaZulu-Natal. A qualitative approach with exploratory, descriptive and contextual strategies was used in this research. Semi-structured interviews were conducted with teachers who had been selected through purposive sampling. Tesch's steps of systematic open coding were used to analyse all of the interviews. The results that emerged during the data analysis indicate that firstly, teachers' experiences of having pregnant learners in their classrooms are negative as these learners are frequently absent from school, perform badly and drop out of school. Secondly, teachers' experiences related to unexpected deliveries are coloured by their lack of the necessary skills and competencies to deal and their resultant insecurity. Thirdly, the teachers feel that they do not receive the assistance they need to deal with teenage pregnancies as well as unexpected deliveries. Collaboration between and continuous support from the Department of Health and the Department of Education are crucial if teenage pregnancies at secondary schools are to be handled with greater success. Support from health care personnel should include guidance on contraception and health campaigns should target both the teachers and secondary school learners. Furthermore, coping strategies for the teachers should form part of the preservice curriculum of teachers.


Structural codes and patient safety: does strict compliance make sense?
The authors of this Comment note recent trends rigidifying the enforcement of building and safety codes for health care facilities and compare the estimated costs (in terms of dollars spent) of those trends with their anticipated benefits (in terms of potential years of human life saved). They estimate that for each potential year of life saved, strict enforcement of the Life Safety Code of the National Fire Protection Association would cost $12.7 to $63.5 million for hospitals and $1.1 to $2.6 million for nursing homes, the latter figure based on Massachusett's experience. These figures are contrasted to the cost of routine kidney dialysis, which is generally acknowledged to be an extremely expensive technology, costing approximately $20,000 per potential year of life saved. The authors suggest that even if strict enforcement of the Code were fully effective (which, given the current structure of the Code, seems doubtful), a portion of the substantial financial resources expended from our limited national health care budget in hewing to the letter of the Code might be better spent on other activities with greater potential yield in improving the quality of life for patients in hospitals and nursing homes.


Anisotropic regulation of Ankrd2 gene expression in skeletal muscle by mechanical stretch.
The diaphragm muscles in vivo are subjected to mechanical forces both in the direction of the muscle fibers and in the direction transverse to the fibers. However, the effect of directional mechanical forces in skeletal muscle gene regulation is completely unknown. Here, we identified that stretch in the longitudinal and transverse directions to the diaphragm muscle fibers up-regulated Ankrd2 gene expression by two distinct signaling pathways in wild-type (WT) and mdm, a mouse model of muscular dystrophy with early-onset of progressive muscle-wasting. Stretch in the longitudinal direction activated both NF-kappaB and AP-1 transcription factors, whereas stretch in the transverse direction activated only AP-1 transcription factor. Interestingly, longitudinal stretch activated Ankrd2 promoter only by NF-kappaB, whereas transverse stretch activated Ankrd2 promoter by AP-1. Moreover, we found that longitudinal stretch activated Akt, which up-regulated Ankrd2 expression through NF-kappaB. However, transverse stretch activated Ras-GTP, Raf-1, and Erk1/2 proteins, which up-regulated Ankrd2 expression through AP-1. Surprisingly, the stretch-activated NF-kappaB and AP-1 signaling pathways was not involved in Ankrd2 regulation at the basal level, which was high in the mdm mouse diaphragm. Taken together, our data show the anisotropic regulation of Ankrd2 gene expression in the diaphragm muscles of WT and mdm mice via two distinct mechanosensitive signaling pathways.


Mechanical occlusion of the inferior vena cava: an early complication after repair of pectus excavatum using the Nuss procedure.
The Nuss procedure is the most widely used surgical procedure to correct pectus excavatum. Although it is a minimally invasive approach, a number of major early complications, such as heart perforation, have been reported. We describe a 15-year-old boy in whom acute occlusion of the inferior vena cava developed after a Nuss repair. The diagnosis was confirmed by emergency postoperative CT examination, and treatment consisted of immediate removal of the Nuss bar.


Synaptic plasticity in the mesolimbic dopamine system.
Long-term potentiation (LTP) and long-term depression (LTD) are thought to be critical mechanisms that contribute to the neural circuit modifications that mediate all forms of experience-dependent plasticity. It has, however, been difficult to demonstrate directly that experience causes long-lasting changes in synaptic strength and that these mediate changes in behaviour. To address these potential functional roles of LTP and LTD, we have taken advantage of the powerful in vivo effects of drugs of abuse that exert their behavioural effects in large part by acting in the nucleus accumbens (NAc) and ventral tegmental area (VTA); the two major components of the mesolimbic dopamine system. Our studies suggest that in vivo drugs of abuse such as cocaine cause long-lasting changes at excitatory synapses in the NAc and VTA owing to activation of the mechanisms that underlie LTP and LTD in these structures. Thus, administration of drugs of abuse provides a distinctive model for further investigating the mechanisms and functions of synaptic plasticity in brain regions that play important roles in the control of motivated behaviour, and one with considerable practical implications.


Study on Microstructure and Mechanical Properties of Laser Welded Dissimilar Joint of P91 Steel and INCOLOY 800HT Nickel Alloy.
This investigation attempts to explore the weld characteristics of a laser welded dissimilar joint of ferritic/martensitic 9Cr-1Mo-V-Nb (P91) steel and Incoloy 800HT austenitic nickel alloy. This dissimilar joint is essential in power generating nuclear and thermal plants operating at 600-650 °C. In such critical operating conditions, it is essential for a dissimilar joint to preserve its characteristics and be free from any kind of defect. The difference between the physical properties of P91 and Incoloy 800HT makes their weldability challenging. Thus, the need for detailed characterization of this dissimilar weld arises. The present work intends to explore the usage of an unconventional welding process (i.e., laser beam welding) and its effect on the joint's characteristics. The single-pass laser welding technique was employed to obtain maximum penetration through the keyhole mode. The welded joint morphology and mechanical properties were studied in as-welded (AW) and post-weld heat treatment (PWHT) conditions. The macro-optical examination shows the complete penetrations with no inclusion and porosities in the weld. The microstructural study was done in order to observe the precipitation and segregation of elements in dendritic and interface regions. Solidification cracks were observed in the weld fusion zone, confirming the susceptibility of Incoloy 800HT to such cracks due to a mismatch between the melting point and thermal conductivity of the base metals. Failure from base metal was observed in tensile test results of standard AW specimen with a yield stress of 265 MPa, and after PWHT, the value increased to 297 MPa. The peak hardness of 391 HV was observed in the P91 coarse grain heat-affected zone (CGHAZ), and PWHT confirmed the reduction in hardness. The impact toughness results that were obtained were inadequate, as the maximum value of impact toughness was obtained for AW P91 heat-affected zone (HAZ) 108 J and the minimum for PWHT Incoloy 800HT HAZ 45 J. Thus, difficulty in obtaining a dissimilar joint with Incoloy 800HT using the laser beam welding technique was observed due to its susceptibility to solidification cracking.


Inhibition of the cardiac Na⁺ channel α-subunit Nav1.5 by propofol and dexmedetomidine.
Propofol and dexmedetomidine are very commonly used sedative agents. However, several case reports demonstrated cardiovascular adverse effects of these two sedatives. Both substances were previously demonstrated to quite potently inhibit neuronal voltage-gated Na(+) channels. Thus, a possible molecular mechanism for some of their cardiac side effects is an inhibition of cardiac voltage gated Na(+) channels. In this study, we therefore explored the effects of propofol and dexmedetomidine on the cardiac predominant Na(+) channel α-subunit Nav1.5. Effects of propofol and dexmedetomidine were investigated on constructs of the human α-subunit Nav1.5 stably expressed in HEK-293 cells by means of whole-cell patch clamp recordings. Both agents induced a concentration-dependent tonic inhibition of Nav1.5. The calculated IC50 value for propofol was 228 ± 10 μM, and for dexmedetomidine 170 ± 20 μM. Tonic block only marginally increased on inactivated channels, and a weak use-dependent block at 10 Hz was observed for dexmedetomidine (16 ± 2 % by 100 μM). The voltage dependencies of fast and slow inactivation as well as the time course of recovery from inactivation were shifted by both propofol and dexmedetomidine. Propofol (IC50 126 ± 47 μM) and dexmedetomidine (IC50 182 ± 27 μM) blocked the persistent sodium current induced by veratradine. Finally, the local-anesthetic (LA)-insensitive mutant Nav1.5-F1760A exhibited reduced tonic and use-dependent block by both substances. Dexmedetomidine was generally more potent as compared to propofol. Propofol and dexmedetomidine seem to interact with the LA-binding site to inhibit the cardiac Na(+) channel Nav1.5 in a state-dependent manner. These data suggest that Nav1.5 is a hitherto unrecognized molecular component of some cardiovascular side effects of these sedative agents.


The Effectiveness of Hospital in Motion, a Multidimensional Implementation Project to Improve Patients' Movement Behavior During Hospitalization.
Hospital in Motion is a multidimensional implementation project aiming to improve movement behavior during hospitalization. The purpose of this study was to investigate the effectiveness of Hospital in Motion on movement behavior. This prospective study used a pre-implementation and post-implementation design. Hospital in Motion was conducted at 4 wards of an academic hospital in the Netherlands. In each ward, multidisciplinary teams followed a 10-month step-by-step approach, including the development and implementation of a ward-specific action plan with multiple interventions to improve movement behavior. Inpatient movement behavior was assessed before the start of the project and 1 year later using a behavioral mapping method in which patients were observed between 9:00 am and 4:00 pm. The primary outcome was the percentage of time spent lying down. In addition, sitting and moving, immobility-related complications, length of stay, discharge destination home, discharge destination rehabilitation setting, mortality, and 30-day readmissions were investigated. Differences between pre-implementation and post-implementation conditions were analyzed using the chi-square test for dichotomized variables, the Mann Whitney test for non-normal distributed data, or independent samples t test for normally distributed data. Patient observations demonstrated that the primary outcome, the time spent lying down, changed from 60.1% to 52.2%. For secondary outcomes, the time spent sitting increased from 31.6% to 38.3%, and discharges to a rehabilitation setting reduced from 6 (4.4%) to 1 (0.7%). No statistical differences were found in the other secondary outcome measures. The implementation of the multidimensional project Hospital in Motion was associated with patients who were hospitalized spending less time lying in bed and with a reduced number of discharges to a rehabilitation setting. Inpatient movement behavior can be influenced by multidimensional interventions. Programs implementing interventions that specifically focus on improving time spent moving, in addition to decreasing time spent lying, are recommended.


Hepatoprotective Actions of Ascorbic Acid, Alpha Lipoic Acid and Silymarin or Their Combination Against Acetaminophen-Induced Hepatotoxicity in Rats.
Background and objectives: Ascorbic acid, alpha lipoic acid (ALA) and silymarin are well-known antioxidants that have hepatoprotective effects. This study aims to investigate the effects of these three compounds combined with attenuating drug-induced oxidative stress and cellular damage, taking acetaminophen (APAP)-induced toxicity in rats as a model both in vivo and in vitro. Materials and Methods: Freshly cultured primary rat hepatocytes were treated with ascorbic acid, ALA, silymarin and their combination, both with and without the addition of APAP to evaluate their in vitro impact on cell proliferation and mitochondrial activity. In vivo study was performed on rats supplemented with the test compounds or their combination for one week followed by two toxic doses of APAP. Results: Selected liver function tests and oxidative stress markers including superoxide dismutase (SOD), malondialdehyde (MDA) and oxidized glutathione (GSSG) were detected. The in vivo results showed that all three pretreatment compounds and their combination prevented elevation of SOD and GSSG serum levels indicating a diminished burden of oxidative stress. Moreover, ascorbic acid, ALA and silymarin in combination reduced serum levels of liver enzymes; however, silymarin markedly maintained levels of all parameters to normal ranges. Silymarin either alone or combined with ascorbic acid and ALA protected cultured rat hepatocytes and increased cellular metabolic activity. The subjected agents were capable of significantly inhibiting the presence of oxidative stress induced by APAP toxicity and the best result for protection was seen with the use of silymarin. Conclusions: The measured liver function tests may suggest an augmented hepatoprotection of the combination preparation than when compared individually.


Genome-wide linkage analysis of 723 affected relative pairs with late-onset Alzheimer's disease.
Previous attempts to identify genetic loci conferring risk for late-onset Alzheimer's disease (LOAD) through linkage analysis have observed some regions of linkage in common. However, due to the sometimes-considerable overlap between the samples, some of these reports cannot be considered to be independent replications. In order to assess the strength of the evidence for linkage and to obtain the best indication of the location of susceptibility genes, we have amalgamated three large samples to give a total of 723 affected relative pairs (ARPs). Multipoint, model-free ARP linkage analysis was performed. Genome-wide significant evidence for linkage was observed on 10q21.2 (LOD=3.3) and genome-wide suggestive evidence was observed on 9q22.33 (LOD=2.5) and 19q13.32 (LOD=2.0). One further region on 9p21.3 was identified with an LOD score>1. We observe no evidence to suggest that more than one locus is responsible for the linkage to 10q21.2, although this linked region may harbour more than one susceptibility gene. Evidence of allele-sharing heterogeneity between the original collection sites was observed on chromosome 9 but not on chromosome 10 or 19. Evidence for an interaction was observed between loci on chromosomes 10 and 19. Where samples overlapped, the genotyping consistency was high, estimated to average at 97.3%. Our large-scale linkage analysis consolidates clear evidence for a susceptibility locus for LOAD on 10q21.2.


High sodium intake increases HCO(3)- absorption in medullary thick ascending limb through adaptations in basolateral and apical Na+/H+ exchangers.
A high sodium intake increases the capacity of the medullary thick ascending limb (MTAL) to absorb HCO(3)(-). Here, we examined the role of the apical NHE3 and basolateral NHE1 Na(+)/H(+) exchangers in this adaptation. MTALs from rats drinking H(2)O or 0.28 M NaCl for 5-7 days were perfused in vitro. High sodium intake increased HCO(3)(-) absorption rate by 60%. The increased HCO(3)(-) absorptive capacity was mediated by an increase in apical NHE3 activity. Inhibiting basolateral NHE1 with bath amiloride eliminated 60% of the adaptive increase in HCO(3)(-) absorption. Thus the majority of the increase in NHE3 activity was dependent on NHE1. A high sodium intake increased basolateral Na(+)/H(+) exchange activity by 89% in association with an increase in NHE1 expression. High sodium intake increased apical Na(+)/H(+) exchange activity by 30% under conditions in which basolateral Na(+)/H(+) exchange was inhibited but did not change NHE3 abundance. These results suggest that high sodium intake increases HCO(3)(-) absorptive capacity in the MTAL through 1) an adaptive increase in basolateral NHE1 activity that results secondarily in an increase in apical NHE3 activity; and 2) an adaptive increase in NHE3 activity, independent of NHE1 activity. These studies support a role for NHE1 in the long-term regulation of renal tubule function and suggest that the regulatory interaction whereby NHE1 enhances the activity of NHE3 in the MTAL plays a role in the chronic regulation of HCO(3)(-) absorption. The adaptive increases in Na(+)/H(+) exchange activity and HCO(3)(-) absorption in the MTAL may play a role in enabling the kidneys to regulate acid-base balance during changes in sodium and volume balance.


Target patterns induced by fixed nanoparticles in block copolymer films.
It is well-known that thin films of cylinder-forming block copolymers (BCP) can exhibit a transition from a perpendicular to a parallel cylinder orientation with respect to the supporting solid substrate upon varying film thickness. We show that wave-like oscillations between these morphologies can be induced through the introduction of nanoparticles (NP) into flow-coated and annealed BCP films where the particles span the film thickness and are fixed by irreversible adsorption to the supporting substrate. We hypothesize that these novel "target" patterns arise from residual stresses that build up in the film while undergoing thermal treatment and film formation, and we support this hypothesis by showing the suppression of this type of pattern formation in films that are first thermally annealed near their glass transition T(g) to relax residual stress. Similar undulating height patterns are also observed in heated homopolymer films with nanoparticles, consistent with our thermally induced stress hypothesis of the target pattern formation in BCP films and pointing to the general nature of wave-like thermally induced height variations in heated heterogeneous polymer films. Similar wave patterns should be induced by lithographically etched substrate patterns arising in device fabrication using BCP materials, which makes the phenomena of technological interest. These target patterns also potentially provide valuable information about the presence of residual stresses in cast films that arise during their processing.


Sirenomelia (Mermaid Syndrome): A Case Report.
Sirenomelia, which is also known as mermaid syndrome and characterized by the fusion of the lower extremities, is the most severe form of caudal regression syndrome and one of the rare and lethal congenital malformations. The anomalies that might be seen in this syndrome include pelvic-sacral dysplasia, genital anomalies, bilateral pelvic renal fusion accompanied by renal dysplasia, colon atresia, unilateral umbilical artery, and imperforated anus. The incidence of sirenomelia is 0.8-1 cases in 60,000-100,000 deliveries and the male/female ratio is 2.7-3:1. The case reported in the present study was a 13-week-old male fetus 30 g in weight with a macerated appearance. The upper extremities had a relatively normal appearance but the lower extremities were conjoined and there was a single lower extremity consisting of conjoined feet and toes. In the face, the nasal bridge was sunken, the ears had a low position, and there were cleft palate and cleft lip. Examination of the external genital organs revealed that the penile part was in the anal region. There was no anus opening. The crown-rump length was 8.5cm, the heel-toe length was approx. 1cm, and the rump-heel length was approx. 3.7cm. There were none of the two kidneys, ureter, bladder, urethra, or rectum. In the umbilical cord, there were 2 venous structures, one of which was the artery. Perivillous congestion and hyperemia, perivillous calcification, deciduitis, and focal infarct regions were observed in placental tissues. This report aims to discuss this very rare case together with the literature.


Modified stitching algorithm for annular subaperture stitching interferometry for aspheric surfaces.
In this paper, a modified stitching algorithm for annular subaperture stitching interferometry (ASSI) for aspheric surfaces is proposed. The mathematical model of adjustment error is deduced based on the wavefront aberration theory and rigid body movement; meanwhile, its basic principle and theory are introduced. The modified stitching algorithm is established based on the mathematical model and the simultaneous least-squares method, which keeps the error from transmitting and accumulating. So the adjustment error can be compensated efficiently. In addition, the standard deviation (SD) in the overlapped regions is used as the figure of merit to determine the stitching accuracy. Finally, simulations and experiments are given to verify the validity and rationality of the proposed algorithm. The results show that the introduced method is quite efficient.


Quantitative sensory testing of intraoral open wounds.
Wound healing is an important aspect of oral and maxillofacial surgery. Positive sensory signs (allodynia, hyperalgesia) and negative sensory signs (hypoesthesia, hypoalgesia) may be encountered. Quantitative sensory testing (QST) has moved from bench to bedside for the detection, therapy selection and monitoring the recovery of individuals with sensory disturbances. Tracking somatosensory changes during normal and abnormal wound healing has not previously been reported. This report presents data obtained by a novel, automated, non-contact psychophysical method for assessment of wound sensitivity after standardized oral mucosal biopsy. By directing graded air puffs towards palatal biopsy wounds, thresholds for sensory detection, pain detection and pain tolerance were repeatedly assessed across 19 days, demonstrating high reliability. Participants recorded daily spontaneous and chewing-evoked maximum pains. Pain detection and tolerance thresholds increased linearly across time. Comparison between air puff evoked pain detection threshold and chewing-evoked pain demonstrated a strong correlation. Thus, for the first time, this study tracked the time course of somatosensory sensitivity of wounds induced by oral biopsies. The psychophysical data on wound healing obtained by this automated, contact-free stimulation method can be utilized as a surrogate marker for clinical pain improvements and standardized assessment of intraoral pain sensitivity, for example in oral mucositis.


A systematic review of platinum and taxane resistance from bench to clinic: an inverse relationship.
We undertook a systematic review of the pre-clinical and clinical literature for studies investigating the relationship between platinum and taxane resistance. Medline was searched for (1) cell models of acquired drug resistance reporting platinum and taxane sensitivities and (2) clinical trials of platinum or taxane salvage therapy in ovarian cancer. One hundred and thirty-seven models of acquired drug resistance were identified. 68.1% of cisplatin-resistant cells were sensitive to paclitaxel and 66.7% of paclitaxel-resistant cells were sensitive to cisplatin. A similar inverse pattern was observed for cisplatin vs. docetaxel, carboplatin vs. paclitaxel and carboplatin vs. docetaxel. These associations were independent of cancer type, agents used to develop resistance and reported mechanisms of resistance. Sixty-five eligible clinical trials of paclitaxel-based salvage after platinum therapy were identified. Studies of single agent paclitaxel in platinum-resistant ovarian cancer where patients had previously recieved paclitaxel had a pooled response rate of 35.3%, n=232, compared to 22% in paclitaxel naïve patients n=1918 (p<0.01, Chi-squared). Suggesting that pre-treatment with paclitaxel may improve the response of salvage paclitaxel therapy. The response rate to paclitaxel/platinum combination regimens in platinum-sensitive ovarian cancer was 79.5%, n=88 compared to 49.4%, n=85 for paclitaxel combined with other agents (p<0.001, Chi-squared), suggesting a positive interaction between taxanes and platinum. Therefore, the inverse relationship between platinum and taxanes resistance seen in cell models is mirrored in the clinical response to these agents in ovarian cancer. An understanding of the cellular and molecular mechanisms responsible would be valuable in predicting response to salvage chemotherapy and may identify new therapeutic targets.


Anticholinergic Medication Use and Fractures in Postmenopausal Women: Findings from the Women's Health Initiative.
Anticholinergic medication use has been associated with several negative health outcomes in older adults, but little is known about its risk for fractures. To examine the association between anticholinergic medication use and fracture outcomes in community-dwelling postmenopausal women. Utilizing a prospective cohort design, we examined data collected from 137,408 women aged 50-79 years from the Women's Health Initiative observational study and clinical trials not reporting hip fracture at baseline. Medications with moderate or strong anticholinergic effects were identified directly from drug containers during in-person interviews. The main outcome was fractures (hip, lower arm/wrist and total fractures). We performed multivariable Cox proportional hazard survival modelling to assess the association between anticholinergic use and the risk of fractures. At baseline, 10.6% of the women were using an anticholinergic medication, of which antihistamines were the most common medication class (48.4%). The multivariable-adjusted hazard ratios for anticholinergic medication use were 1.08 (95% confidence interval [CI] 0.89-1.30) for hip fracture, 1.01 (95% CI 0.91-1.13) for lower arm/wrist fracture and 1.03 (95% CI 0.98-1.09) for total fractures. We observed no association according to subclass or count of anticholinergic medications, or trends between the duration of anticholinergic use and any of the fracture outcomes. Anticholinergic medication use was not associated with an increased risk of fractures among community-dwelling women. Future research should make efforts to capture over-the-counter medication use and the cumulative anticholinergic burden in relation to important health outcomes for older adults.


An overall view of temperature oscillations in the solar chromosphere with ALMA.
By direct measurements of the gas temperature, the Atacama Large Millimeter/submillimeter Array (ALMA) has yielded a new diagnostic tool to study the solar chromosphere. Here, we present an overview of the brightness-temperature fluctuations from several high-quality and high-temporal-resolution (i.e. 1 and 2 s cadence) time series of images obtained during the first 2 years of solar observations with ALMA, in Band 3 and Band 6, centred at around 3 mm (100 GHz) and 1.25 mm (239 GHz), respectively. The various datasets represent solar regions with different levels of magnetic flux. We perform fast Fourier and Lomb-Scargle transforms to measure both the spatial structuring of dominant frequencies and the average global frequency distributions of the oscillations (i.e. averaged over the entire field of view). We find that the observed frequencies significantly vary from one dataset to another, which is discussed in terms of the solar regions captured by the observations (i.e. linked to their underlying magnetic topology). While the presence of enhanced power within the frequency range 3-5 mHz is found for the most magnetically quiescent datasets, lower frequencies dominate when there is significant influence from strong underlying magnetic field concentrations (present inside and/or in the immediate vicinity of the observed field of view). We discuss here a number of reasons which could possibly contribute to the power suppression at around 5.5 mHz in the ALMA observations. However, it remains unclear how other chromospheric diagnostics (with an exception of Hα line-core intensity) are unaffected by similar effects, i.e. they show very pronounced 3-min oscillations dominating the dynamics of the chromosphere, whereas only a very small fraction of all the pixels in the 10 ALMA datasets analysed here show peak power near 5.5 mHz. This article is part of the Theo Murphy meeting issue 'High-resolution wave dynamics in the lower solar atmosphere'.


Why is the placenta being ignored?
The relationship between the frequency of published recommended indications for placental pathological examination and the frequency of requests for such examination in a population-based study of term newborn encephalopathy was examined. Only 11.2% of placentas among 276 case infants and 0.7% of placentas among 564 term control infants were examined. Using the criteria set out in a consensus statement by the American College of Pathologists, all 276 cases fulfilled multiple maternal, fetal and placental indications for placental examination. Furthermore 43.3% of control infants fulfilled at least one criterion. Of the 25 case placentas that underwent pathological review, 16 were reported as having no diagnostic abnormality Six cases (24%) showed clinically important findings: four had evidence of infection, one had multiple chorangiomata and one had thrombosis and rupture of the umbilical vein. Of the three remaining placentas, one showed funisitis, one showed minor lymphohistiocytic villitis and one was from monochorionic twins. To our knowledge there are no agreed Australian guidelines for when a placenta should be submitted for pathological examination. We suggest that until guidelines based on properly designed studies are developed it may be appropriate to store all placentas for at least 72 hours. If the infant develops neurological symptoms or requires unexpected admission to a neonatal intensive care unit then placental examination may reveal important aetiological diagnostic and prognostic information.


Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation.
While the catalog of mammalian transcripts and their expression levels in different cell types and disease states is rapidly expanding, our understanding of transcript function lags behind. We present a robust technology enabling systematic investigation of the cellular consequences of repressing or inducing individual transcripts. We identify rules for specific targeting of transcriptional repressors (CRISPRi), typically achieving 90%-99% knockdown with minimal off-target effects, and activators (CRISPRa) to endogenous genes via endonuclease-deficient Cas9. Together they enable modulation of gene expression over a ∼1,000-fold range. Using these rules, we construct genome-scale CRISPRi and CRISPRa libraries, each of which we validate with two pooled screens. Growth-based screens identify essential genes, tumor suppressors, and regulators of differentiation. Screens for sensitivity to a cholera-diphtheria toxin provide broad insights into the mechanisms of pathogen entry, retrotranslocation and toxicity. Our results establish CRISPRi and CRISPRa as powerful tools that provide rich and complementary information for mapping complex pathways.


The case against ergocalciferol (vitamin D2) as a vitamin supplement.
Supplemental vitamin D is available in 2 distinct forms: ergocalciferol (vitamin D2) and cholecalciferol (vitamin D3). Pharmacopoeias have officially regarded these 2 forms as equivalent and interchangeable, yet this presumption of equivalence is based on studies of rickets prevention in infants conducted 70 y ago. The emergence of 25-hydroxyvitamin D as a measure of vitamin D status provides an objective, quantitative measure of the biological response to vitamin D administration. As a result, vitamin D3 has proven to be the more potent form of vitamin D in all primate species, including humans. Despite an emerging body of evidence suggesting several plausible explanations for the greater bioefficacy of vitamin D3, the form of vitamin D used in major preparations of prescriptions in North America is vitamin D2. The case that vitamin D2 should no longer be considered equivalent to vitamin D3 is based on differences in their efficacy at raising serum 25-hydroxyvitamin D, diminished binding of vitamin D2 metabolites to vitamin D binding protein in plasma, and a nonphysiologic metabolism and shorter shelf life of vitamin D2. Vitamin D2, or ergocalciferol, should not be regarded as a nutrient suitable for supplementation or fortification.


Schizotypy and mental time travel.
Mental time travel is the capacity to imagine the autobiographical past and future. Schizotypy is a dimensional measure of psychosis-like traits found to be associated with creativity and imagination. Here, we examine the phenomenological qualities of mental time travel in highly schizotypal individuals. After recollecting past episodes (autobiographical memory) and imagining future events (episodic future thinking), those scoring highly on positive schizotypy reported a greater sense of 'autonoetic awareness,' defined as a greater feeling of mental time travel and re-living/'pre-living' imagined events. Furthermore, in contrast to other sensory domains, imagery of the past and future episodes contained more olfactory detail in these high scorers. The results are discussed in relation to previous reports of anomalous olfactory experiences in schizotypy and heightened vividness of olfactory imagery in post-traumatic stress disorder, for which schizotypy is a risk factor.


The alpha-glucosidase inhibitor N-butyldeoxynojirimycin inhibits human immunodeficiency virus entry at the level of post-CD4 binding.
The alpha-glucosidase inhibitor N-butyldeoxynojirimycin (NB-DNJ) is a potent inhibitor of human immunodeficiency virus (HIV) replication and syncytium formation in vitro. However, the exact mechanism of action of NB-DNJ remains to be determined. In this study we have examined the impairment of HIV infectivity mediated by NB-DNJ. By two independent HIV entry assays [PCR-based HIV entry assay and entry of Cocal(HIV) pseudotypes], the reduction in infectivity was found to be due to an impairment of viral entry. No effect of NB-DNJ treatment was seen on the kinetics of the interaction between gp120 and CD4 (surface plasmon resonance; BIAcore) or on the binding of virus particles to H9 cells (using radiolabeled virions). We therefore conclude that a major mechanism of action of NB-DNJ as an inhibitor of HIV replication is the impairment of viral entry at the level of post-CD4 binding, due to an effect on viral envelope components.


Tumor metabolism of malignant gliomas.
Constitutively activated oncogenic signaling via genetic mutations such as in the EGFR/PI3K/Akt and Ras/RAF/MEK pathways has been recognized as a major driver for tumorigenesis in most cancers. Recent insights into tumor metabolism have further revealed that oncogenic signaling pathways directly promote metabolic reprogramming to upregulate biosynthesis of lipids, carbohydrates, protein, DNA and RNA, leading to enhanced growth of human tumors. Therefore, targeting cell metabolism has become a novel direction for drug development in oncology. In malignant gliomas, metabolism pathways of glucose, glutamine and lipid are significantly reprogrammed. Moreover, molecular mechanisms causing these metabolic changes are just starting to be unraveled. In this review, we will summarize recent studies revealing critical gene alterations that lead to metabolic changes in malignant gliomas, and also discuss promising therapeutic strategies via targeting the key players in metabolic regulation.


Predictive value of scintigraphic (semi-)quantitative thyroid parameters on radioiodine therapy outcome in hyperthyroid cats.
Objectives The outcome of radioiodine therapy in hyperthyroid cats is suspected to be influenced by multiple factors. The degree of activity of the thyroid gland, represented by uptake of sodium pertechnetate or tracer activities of radioiodine by the thyroid gland on thyroid scintigraphy, has been suggested in the literature as one of those. Thyroid gland pertechnetate uptake can be represented by (semi-)quantitative factors such as the thyroid to salivary gland (T/S) ratio, the thyroid to background (T/B) ratio and the percentage technetium uptake by the thyroid glands (%TcU). The aim of this study was to investigate a possible relationship between these thyroid scan parameters and radioiodine therapy outcome. Methods Sodium pertechnetate thyroid scans of 75 hyperthyroid cats were retrospectively evaluated and statistical analysis was performed with and without correction for injected radioiodine activity. Three different background regions of interest (ROIs) were used to calculate the T/B ratio and %TcU: 'neck', 'circle' and 'copy ROI'. Results Higher T/S ratios were found to be significantly related to a persistent hyperthyroid outcome in both analyses. For the T/S ratio, a threshold value of 5.4 was determined, with a sensitivity of 73% and a specificity of 59%. An increased risk for persistent hyperthyroidism compared with a final euthyroid outcome with an increased T/Bcircle ratio was only found to be significant without correction for the activity of radioiodine administered. For the %TcU no statistical significance was reached. Regarding a low total thyroxine outcome, no significant relationships with any of the investigated parameters were found. Conclusions and relevance The findings of this study suggest that semi-quantification of thyroid gland uptake is best performed using the T/S ratio. A T/S ratio ⩾5.4 is a possible indicator for an increased risk of persistent hyperthyroidism.


Susceptibility of oral bacteria to Melaleuca alternifolia (tea tree) oil in vitro.
The in vitro activity of Melaleuca alternifolia (tea tree) oil against 161 isolates of oral bacteria from 15 genera was determined. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) ranged from 0.003 to 2.0% (v/v). MIC90 values were 1.0% (v/v) for Actinomyces spp., Lactobacillus spp., Streptococcus mitis and Streptococcus sanguis, and 0.1% (v/v) for Prevotella spp. Isolates of Porphyromonas, Prevotella and Veillonella had the lowest MICs and MBCs, and isolates of Streptococcus, Fusobacterium and Lactobacillus had the highest. Time kill studies with Streptococcus mutans and Lactobacillus rhamnosus showed that treatment with > or = 0.5% tea tree oil caused decreases in viability of >3 log colony forming units/ml after only 30 s, and viable organisms were not detected after 5 min. These studies indicate that a range of oral bacteria are susceptible to tea tree oil, suggesting that tea tree oil may be of use in oral healthcare products and in the maintenance of oral hygiene.


Neuronal differentiation potential of human adipose-derived mesenchymal stem cells.
Adult mesenchymal stem cells derived from adipose tissue (A-MSC) have the capacity to differentiate in vitro into mesenchymal as well as endodermal and ectodermal cell lineages. We investigated the neuronal differentiation potential of human A-MSC with a protocol which included sphere formation and sequential culture in brain-derived neurotrophic factor (BDNF) and retinoic acid (RA). After 30 days, about 57% A-MSC showed morphological, immunocytochemical and electrophysiological evidence of initial neuronal differentiation. In fact, A-MSC displayed elongated shape with protrusion of two or three cellular processes, selectively expressed nestin and neuronal molecules (including GABA receptor and tyroxine hydroxilase) in the absence of glial phenotypic markers. Differentiated cells showed negative membrane potential (-60 mV), delayed rectifier potassium currents and TTX-sensitive sodium currents. Such changes were stable for at least 7 days after removal of differentiation medium. In view of these results and the easy availability of adipose tissue, A-MSC may be a ready source of adult MSC with neuronal differentiation potential, an useful tool to treat neurodegenerative diseases.


In situ identification of Legionellaceae using 16S rRNA-targeted oligonucleotide probes and confocal laser scanning microscopy.
Bacteria of the family Legionellaceae form a monophyletic group within the gamma-subclass of Proteobacteria. Based on comparative sequence analysis we constructed two oligonucleotide probes complementary to regions of 16S rRNA characteristic for Legionellaceae. Probe specificities were tested by whole-cell or dot-blot hybridization against 14 serogroups of Legionella pneumophila, 22 different Legionella spp. and 72 non-legionellae reference strains. Using optimized conditions both probes hybridized to all tested strains of L. pneumophila. Probes LEG226 and LEG705 hybridized to 71% and 90% of the Legionella species tested, respectively. With the exception of Methylomonas alba none of the non-target strains showed complete sequence homology within the target molecule. In a preliminary evaluation the results of classical techniques employing selective media, immunofluorescence and the probe assay were in good accordance for routine environmental and clinical isolates. L. pneumophila suspended in drinking water at approximately 10(3)-10(4) c.f.u. ml-1 could be rapidly detected by a combination of membrane filtration on polycarbonate filters and whole-cell hybridization. Even after incubation for 1 year a proportion of the released cells was still detectable. In situ hybridization also facilitated visualization of Legionella spp, cells in model biofilms. A combination of in situ hybridization and confocal laser scanning microscopy (CLSM) was used to analyse the three-dimensional arrangement of L. pneumophila within cells of the ciliated protozoan Tetrahymena pyriformis. Whole-cell probing with 16S rRNA-targeted oligonucleotides could, in the future, complement established techniques like immunofluorescence and PCR in ecological and epidemiological studies of Legionellaceae.


Identifying tremor-related characteristics of basal ganglia nuclei during movement in the Parkinsonian patient.
Local field potential (LFP) and Electromyographic (EMG) signals were recorded from 12 Parkinsonian patients with tremor-dominant symptoms as they performed passive and voluntary movements. The LFP signals were categorised into episodes of tremorous and atremorous activity (identified through EMG power spectra), then divided into delta (2-4 Hz), theta (4-8 Hz), alpha (8-13 Hz), and beta (13-30 Hz) frequency bands. Modulation of LFP oscillatory activity in these frequency bands were compared between the subthalamic nucleus (STN) and the globus pallidus internus (GPi) to determine if differential tremor-related characteristics were identifiable for either target. Our results suggest that such local characteristic activity is identifiable in the STN, and thus could be a target for initial development of a closed-loop demand driven stimulator device which capitalises on such activity to trigger stimulation, even during voluntary movement activity.


Demonstration of an antibody-mediated tolerance state and its effect on antibody affinity.
We have described a model of immunological tolerance induced, in adult mice, by a single injection of a moderate dose of a hapten-protein conjugate. The data suggest that the mechanism of this tolerance state is the production of small amounts of high affinity antibody in response to the tolerance-inducing antigen injection. This antibody acts to inhibit the response to a subsequent challenge with antigen in complete Freund's adjuvant by a mechanism comparable to that of passive antibody-medicated immune suppression. It was shown that a small but high affinity. Tolerance was not terminated by transfer of normal syngeneic spleen or peritoneal cells into tolerant animals. Spleen cells from tolerant mice, when transferred into lethally irradiated, syngeneic animals, produced a PFC response which is greater in magnitude and tolerance state had a significant degree of carrier specificity which was shown to be comparable to the carrier specificity of antibody-mediated immune suppression. hus, evidence was presented to show that one mechanism of tolerance in adult animals in the suppressive effect of small amounts of high affinity antibody formed in response to the tolerizing injection of antigen.


papA gene of avian pathogenic Escherichia coli.
P fimbrial adhesins may be associated with the virulence of avian pathogenic Escherichia coli (APEC). However, most APECs are unable to express P fimbriae even when they are grown under conditions that favor P fimbrial expression. This failure can be explained by the complete absence of the pap operon or the presence of an incomplete pap operon in Pap-negative APEC strains. In the present study, we analyzed the pap operon, specifically the papA gene that encodes the major fimbrial shaft, to better understand the pap gene cluster at the genetic level. First, by PCR, we examined a collection of 500 APEC strains for the presence of 11 genes comprising the pap operon. Except for papA, all the other genes of the operon were present in 38% to 41.2% of APEC, whereas the papA was present only in 10.4% of the APEC tested. Using multiplex PCR to probe for allelic variants of papA, we sought to determine if the low prevalence of papA among APEC was related to genetic heterogeneity of the gene itself. It was determined that the papA of APEC always belongs to the F11 allelic variant. Finally, we sequenced the 'papA region' from two papA-negative strains, both of which contain all the other genes of the pap operon. Interestingly, both strains had an 11,104-bp contig interruptingpapA at the 281-bp position. This contig harbored a streptomycin resistance gene and a classic Tn10 transposon containing the genes that confer tetracycline resistance. However, we noted that the papA gene of every papA-negative APEC strain was not interrupted by an 11,104-bp contig. It is likely that transposons bearing antibiotic resistance genes have inserted within pap gene cluster of some APEC strains, and such genetic events may have been selected for by antibiotic use.


Differentiation of Human Embryonic Stem Cells to Sympathetic Neurons: A Potential Model for Understanding Neuroblastoma Pathogenesis.
Previous studies modelling human neural crest differentiation from stem cells have resulted in a low yield of sympathetic neurons. Our aim was to optimise a method for the differentiation of human embryonic stem cells (hESCs) to sympathetic neuron-like cells (SN) to model normal human SNS development. Using stromal-derived inducing activity (SDIA) of PA6 cells plus BMP4 and B27 supplements, the H9 hESC line was differentiated to neural crest stem-like cells and SN-like cells. After 7 days of PA6 cell coculture, mRNA expression of SNAIL and SOX-9 neural crest specifier genes and the neural marker peripherin (PRPH) increased. Expression of the pluripotency marker OCT 4 decreased, whereas TP53 and LIN28B expression remained high at levels similar to SHSY5Y and IMR32 neuroblastoma cell lines. A 5-fold increase in the expression of the catecholaminergic marker tyrosine hydroxylase (TH) and the noradrenergic marker dopamine betahydroxylase (DBH) was observed by day 7 of differentiation. Fluorescence-activated cell sorting for the neural crest marker p75, enriched for cells expressing p75, DBH, TH, and PRPH, was more specific than p75 neural crest stem cell (NCSC) microbeads. On day 28 post p75 sorting, dual immunofluorescence identified sympathetic neurons by PRPH and TH copositivity cells in 20% of the cell population. Noradrenergic sympathetic neurons, identified by copositivity for both PHOX2B and DBH, were present in 9.4% ± 5.5% of cells. We have optimised a method for noradrenergic SNS development using the H9 hESC line to improve our understanding of normal human SNS development and, in a future work, the pathogenesis of neuroblastoma.


Visual sensitivity in the presence of a patterned background.
The sensitivity of the visual system depends on ambient illumination: Sensitivity is reduced in the presence of a bright, uniform background. We asked how sensitivity is adjusted when the background is spatially detailed and therefore contains both luminance peaks and troughs in the neighborhood of a foreground object. A test flash was superimposed on a static sinusoidal grating. As the grating's spatial frequency increased, sensitivity for flash detection declined, regardless of whether the flash was superimposed on a peak or a trough of the grating. We studied the mechanisms underlying this loss of sensitivity by delivering the test stimulus through one eye and the background through the other. The conclusion is that three mechanisms are involved. Luminance adaptation and a masking process adjust sensitivity at low- and mid-range spatial frequencies, respectively. The third mechanism, a contrast gain control, is localized (it occurs at spatial frequencies approaching the limit for resolution) and fast (complete in half a second), and it results from early processing in the visual pathway (it is absent during dichoptic viewing). This local adjustment of sensitivity may help to protect the clarity of even the smallest details in the visual scene.


Global transcriptional analysis identifies a novel role for SOX4 in tumor-induced angiogenesis.
The expression of the transcription factor SOX4 is increased in many human cancers, however, the pro-oncogenic capacity of SOX4 can vary greatly depending on the type of tumor. Both the contextual nature and the mechanisms underlying the pro-oncogenic SOX4 response remain unexplored. Here, we demonstrate that in mammary tumorigenesis, the SOX4 transcriptional network is dictated by the epigenome and is enriched for pro-angiogenic processes. We show that SOX4 directly regulates endothelin-1 (ET-1) expression and can thereby promote tumor-induced angiogenesis both in vitro and in vivo. Furthermore, in breast tumors, SOX4 expression correlates with blood vessel density and size, and predicts poor-prognosis in patients with breast cancer. Our data provide novel mechanistic insights into context-dependent SOX4 target gene selection, and uncover a novel pro-oncogenic role for this transcription factor in promoting tumor-induced angiogenesis. These findings establish a key role for SOX4 in promoting metastasis through exploiting diverse pro-tumorigenic pathways.


Synergistic genetic defect in B-lymphocyte function. I. Defective responses to B-cell stimulants and their genetic basis.
CBA/N female mice, which express an X-linked defect in B-lymphocyte function, were mated with C3H/HeJ male mice, which are unresponsive to lipopolysaccharide (LPS). The resulting F1 hybrid females were mated to C3H/HeJ males. Approximately one-half of the backcross (BC.1) males obtained from this mating expressed a more profound immunologic defect than either of the parental strains. Spleen cells from these mice were unresponsive to a series of B-cell mitogens including LPS prepared from Escherichia coli K235 and from E. coli 0111:B4, lipoprotein mitogen from E. coli, and Nocardia water-soluble mitogen (NWSM). They failed to give in vitro antibody responses to the thymus-independent type 2 (TI-2) antigen trinophenylated Ficoll and most were unresponsive to the TI-1 antigens trinitrophenylated Brucella abortus, trinitrophenylated LPS, and trinitrophenylated NWSM. This synergistic defect in B-lymphocyte function depended on the presence of the CBA/N xid gene but the critical gene(s) from the C3H strain was not the defective Lps gene (Lpsd). These mice should provide a valuable tool for the elucidation of B-lymphocyte ontogeny, heterogeneity, and function.


Lack of causal relationship between dentinal microcracks and root canal preparation with reciprocation systems.
This study aimed to evaluate the frequency of dentinal microcracks observed after root canal preparation with 2 reciprocating and a conventional full-sequence rotary system using micro-computed tomographic analysis. Thirty mesial roots of mandibular molars presenting a type II Vertucci canal configuration were scanned at an isotropic resolution of 14.16 μm. The sample was randomly assigned to 3 experimental groups (n = 10) according to the system used for the root canal preparation: group A-Reciproc (VDW, Munich, Germany), group B-WaveOne (Dentsply Maillefer, Baillagues, Switzerland), and group C-BioRaCe (FKG Dentaire, La-Chaux-de-Fonds, Switzerland). Second and third scans were taken after the root canals were prepared with instruments sizes 25 and 40, respectively. Then, pre- and postoperative cross-section images of the roots (N = 65,340) were screened to identify the presence of dentinal defects. Dentinal microcracks were observed in 8.72% (n = 5697), 11.01% (n = 7197), and 7.91% (n = 5169) of the cross-sections from groups A (Reciproc), B (WaveOne), and C (BioRaCe), respectively. All dentinal defects identified in the postoperative cross-sections were also observed in the corresponding preoperative images. No causal relationship between dentinal microcrack formation and canal preparation procedures with Reciproc, WaveOne, and BioRaCe systems was observed.


Flow of suspensions of carbon nanotubes carrying phase change materials through microchannels and heat transfer enhancement.
This work explores the potential of nano-encapsulated phase change materials (PCMs) in applications related to microelectronics cooling. PCMs (wax or meso-erythritol) were encapsulated in carbon nanotubes (CNTs) by a method of self-sustained diffusion at room temperature and pressure. These nano-encapsulated wax nanoparticles alone allowed heat removal over a relatively wide range of temperatures (different waxes have melting temperatures in the range 40-80 °C). On the other hand, nano-encapsulated meso-erythritol nanoparticles allowed heat removal in the range 118-120 °C. The combination of these two PCMs (wax and meso-erythritol) could extend the temperature range to 40-120 °C, when both types of nanoparticles (wax and meso-erythritol intercalated) would be suspended in the same carrier fluid (an oil). The nanoparticles possess a short response time of the order of 10(-7) s. Such nano-encapsulation can also prevent the PCM from sticking to the wall. In this work, experiments with wax-intercalated CNTs, stable aqueous suspensions of CNTs with concentrations up to 3 wt% with and without nano-encapsulated wax were prepared using a surfactant sodium dodecyl benzene sulfonate (NaDDBS). These suspensions were pumped through two channels of 603 μm or 1803 μm in diameter subjected to a constant heat flux at the wall. It was found that the presence of the surfactant in CNT suspensions results in a pseudo-slip at the channel wall which enhances the flow rate at a fixed pressure drop. When aqueous solutions of the surfactant were employed (with no CNTs added), the enhanced convection alone was responsible for a ~2 °C reduction in temperature in comparison with pure water flows. When CNTs with nano-encapsulated wax were added, an additional ~1.90 °C reduction in temperature due to the PCM fusion was observed when using 3 wt% CNT suspensions. In addition, suspensions of meso-erythritol-intercalated CNTs in alpha-olefin oil were used as coolants in flows through the 1803 μm-diameter microchannel. These suspensions (1.5 wt% CNT) revealed a temperature reduction due to the PCM fusion of up to 3.2 °C, and a fusion temperature in the range 118-120 °C.


Association of HPA axis-related genetic variation with stress reactivity and aggressive behaviour in pigs.
Stress, elicited for example by aggressive interactions, has negative effects on various biological functions including immune defence, reproduction, growth, and, in livestock, on product quality. Stress response and aggressiveness are mutually interrelated and show large interindividual variation, partly attributable to genetic factors. In the pig little is known about the molecular-genetic background of the variation in stress responsiveness and aggressiveness. To identify candidate genes we analyzed association of DNA markers in each of ten genes (CRH g.233C>T, CRHR1 c.*866_867insA, CRHBP c.51G>A, POMC c.293_298del, MC2R c.306T>G, NR3C1 c.*2122A>G, AVP c.207A>G, AVPR1B c.1084A>G, UCN g.1329T>C, CRHR2 c.*13T>C) related to the hypothalamic-pituitary-adrenocortical (HPA) axis, one of the main stress-response systems, with various stress- and aggression-related parameters at slaughter. These parameters were: physiological measures of the stress response (plasma concentrations of cortisol, creatine kinase, glucose, and lactate), adrenal weight (which is a parameter reflecting activity of the central branch of the HPA axis over time) and aggressive behaviour (measured by means of lesion scoring) in the context of psychosocial stress of mixing individuals with different aggressive temperament. The SNP NR3C1 c.*2122A>G showed association with cortisol concentration (p = 0.024), adrenal weight (p = 0.003) and aggressive behaviour (front lesion score, p = 0.012; total lesion score p = 0.045). The SNP AVPR1B c.1084A>G showed a highly significant association with aggressive behaviour (middle lesion score, p = 0.007; total lesion score p = 0.003). The SNP UCN g.1329T>C showed association with adrenal weight (p = 0.019) and aggressive behaviour (front lesion score, p = 0.029). The SNP CRH g.233C>T showed a significant association with glucose concentration (p = 0.002), and the polymorphisms POMC c.293_298del and MC2R c.306T>G with adrenal weight (p = 0.027 and p < 0.0001 respectively). The multiple and consistent associations shown by SNP in NR3C1 and AVPR1B provide convincing evidence for genuine effects of their DNA sequence variation on stress responsiveness and aggressive behaviour. Identification of the causal functional molecular polymorphisms would not only provide markers useful for pig breeding but also insight into the molecular bases of the stress response and aggressive behaviour in general.


BARIATRIC SURGERY IN BRAZILIAN PUBLIC HEALTH SYSTEM: THE GOOD, THE BAD AND THE UGLY, OR A LONG WAY TO GO. YELLOW SIGN!
In Brazil, there has been a significant increase in obesity rates in all age groups. Data from 2017 show that obesity affects 19% of the population. Due to the magnitude of the problem, public health policies have aimed to prevent complications related to obesity by increasing the offerfor bariatric surgeries. To analyze the current status of bariatric surgery performed in the Brazilian public health system, including data from macroregions and also the effect of digestive surgery training on the number of procedures. The database of the public health registry (DATASUS) was assessedbetween 2008 and 2018 for descriptive analysis of data and evaluation of the selected parameters. The main surgical techniques, comorbidities, mortality and the costs profile of the system were evaluated. There was a 339% increase in the number of bariatric surgeries in the period evaluated. Gastric bypass was performed in 94% of cases whereas sleeve in 2.4%. Other techniques were used in 3.6%. There were discrepancies in the number of surgeries performed in different regions of the country. There was a considerable advance in the number of bariatric surgeries performed by the public health system between 2008 and 2018. However, there is a need to increase the offer of this service and alsospecialized training, as well as a correction in the distribution of these procedures in the national territory to achieve integrality among its users.


Cloning of an aquaporin homologue present in water channel containing endosomes of toad urinary bladder.
Regulation of total body water balance in amphibians by antidiuretic hormone (ADH) contributed to their successful colonization of terrestrial habitats approximately 200-300 million years ago. In the mammalian kidney, ADH modulates epithelial cell apical membrane water permeability (Pf) by fusion and retrieval of cytoplasmic vesicles containing water channel proteins called aquaporins (AQPs). To determine the role of AQPs in ADH-elicited Pf in amphibians, we have identified and characterized a unique AQP from Bufo marinus called AQP toad bladder (AQP-TB). AQP-TB possesses many structural features common to other AQPs, AQP-TB is expressed abundantly in ADH-responsive tissues, including toad urinary bladder and skin as well as lung, skeletal muscle, kidney, and brain. In a manner identical to that reported for the mammalian ADH-elicited water channel AQP2, AQP-TB expression is increased significantly by intervals of dehydration or chronic ADH stimulation. However, expression of AQP-TB protein in Xenopus laevis oocytes does not significantly increase oocyte Pf. The lack of expression of functional AQP-TB water channels in oocytes may result from intracellular sequestration of AQP-TB due to the presence of a YXRF sequence motif present in its carboxyterminal domain.


Inhibitory effect of heparin on herpes simplex virus.
Nahmias, André J. (Boston University School of Medicine, Boston, Mass.), and Sidney Kibrick. Inhibitory effect of heparin on herpes simplex virus. J. Bacteriol. 87:1060-1066. 1964.-A substance inhibitory to herpes simplex virus was observed during experiments with leukocyte cultures. The component in the cultures responsible for this inhibition was identified as heparin. The minimal inhibitory concentration required to inhibit 30 to 300 tcd(50) of the virus in human amnion tissue culture was found to be 1 to 2 units per ml (10 to 20 mug/ml). This effect was confirmed with other strains of herpes simplex virus, other tissue-culture systems, and other media. The inhibitory activity of the heparin was found to be related to the sulfate groupings on the molecule. The effect of heparin appears to be on the virus, rather than on the cell. The virus is not inactivated, however, and the heparin-virus "complex" is readily dissociable on dilution. Heparin was shown to affect viral infection in its earliest phase, probably at the primary electrostatic attachment of virus to cell. The import of these and related observations on common virological laboratory procedures and the possible biological significance of our findings are discussed.


Functional capacity of solid tissue transplants in the brain: evidence for immunological privilege.
The capacity of the mammalian brain to support the physiological function of allografts was assessed in parathyroidectomized Fischer strain rats bearing either isografts or immunogenic DA allografts of parathyroid glands implanted in their cerebral cortices. Established isografts and allografts survived indefinitely in the brain, maintaining normal serum calcium levels, with equal numbers of spontaneous failures (18-21%) in each group. Similarly, both MHC-compatible and incompatible skin allografts survived and were 'functional' at 40-50 days postgrafting as assessed by: continued formation of keratin; the presence of differentiated hair follicles and sebaceous glands; and frequent mitotic figures. No serum alloantibodies were induced by either MHC-incompatible parathyroid glands or skin in this site. However, both types of allografts were promptly rejected or failed to become established in the brains of specifically presensitized hosts. Furthermore, when Fischer hosts with long-established intracerebral DA parathyroid grafts received orthotopic DA skin grafts, their parathyroid grafts were rejected along with first-set rejection of the skin grafts. The tempo of this cellular immune response and the primary alloantibody response that accompanied it indicate that although the intracerebral grafts failed to induce detectable host sensitization or suppression, they remained susceptible to immune effectors. Thus, by using strongly immunogenic, adult tissues, we have established that the rat cerebral cortex is an immunologically privileged site, and the privilege is not dependent on lack of graft immunogenicity or alterations in host responsiveness. Furthermore, Ia+ (possible antigen-presenting) cells were rare in the cortical parenchyma sites used for transplantation though numerous in the choroid plexus of the ventricles and in certain areas of white matter. Therefore, privilege probably reflects deficient graft antigen presentation related to the paucity of Ia+ cells as well as to the brain's poor lymphatic drainage.


State of the art in the treatment of systemic vasculitides.
Anti-neutrophil cytoplasm antibodies (ANCA) are associated with small vessel vasculitides (AASV) affecting the lungs and kidneys. Structured clinical assessment using the Birmingham Vasculitis Activity Score and Vasculitis Damage Index should form the basis of a treatment plan and be used to document progress, including relapse. Severe disease with organ or life threatening manifestations needs cyclophosphamide or rituximab, plus high dose glucocorticoids, followed by lower dose steroid plus azathioprine, or methotrexate. Additional plasmapheresis is effective for very severe disease, reducing dialysis dependence from 60 to 40% in the first year, but with no effect on mortality or long-term renal function, probably due to established renal damage. In milder forms of ANCA-associated vasculitis, methotrexate, leflunomide, or mycophenolate mofetil are effective. Mortality depends on initial severity: 25% in patients with renal failure or severe lung hemorrhage; 6% for generalized non-life threatening AASV but rising to 30-40% at 5 years. Mortality from GPA is four times higher than the background population. Early deaths are due to active vasculitis and infection. Subsequent deaths are more often due to cardiovascular events, infection, and cancer. We need to improve the long-term outcome, by controlling disease activity but also preventing damage and drug toxicity. By contrast, in large vessel vasculitis where mortality is much less but morbidity potentially greater, such as giant cell arteritis (GCA) and Takayasu arteritis, therapeutic options are limited. High dose glucocorticoid results in significant toxicity in over 80%. Advances in understanding the biology of the vasculitides are improving therapies. Novel, mechanism based therapies such as rituximab in AASV, mepolizumab in eosinophilic granulomatosis with polyangiitis, and tocilizumab in GCA, but the lack of reliable biomarkers remains a challenge to progress in these chronic relapsing diseases.


Angiotensin Receptor Neprilysin Inhibitor in Japanese Patients With Heart Failure and Reduced Ejection Fraction - Baseline Characteristics and Treatment of PARALLEL-HF Trial.
The objective of the present analyses was to describe the baseline characteristics and treatment of the Japanese patients with HFrEF in THE PARALLEL-HF study. Methods and Results: Key demographic, clinical and laboratory findings, along with treatment, were reported and compared with patients enrolled in the PARADIGM-HF trial and other contemporary randomized clinical trials and registries of Japanese patients with HFrEF. In addition, the MAGGIC and EMPHASIS-HF risk scores were calculated. A total of 225 Japanese patients were randomized in PARALLEL-HF with a mean age of 67.9 years and the majority of the patients being male (85.8%) and in NYHA Class II (93.8%). Key baseline characteristics in PARALLEL-HF were generally comparable with PARADIGM-HF, and other contemporary clinical trials and registries of Japanese HFrEF patients. Patients enrolled in PARALLEL-HF were well treated with conventional evidence-based therapy at baseline (angiotensin-converting enzyme inhibitor inhibitor/angiotensin receptor blocker, 62.7%/37.3%; β-blockers, 94.7%; mineralocorticoid receptor antagonist, 59.1%). Despite the evidence-based treatment and most patients being in NYHA Class II, these patients had a low LVEF (mean 28.1%) and were at high risk of cardiovascular mortality and morbidity as assessed by the MAGGIC and EMPHASIS-HF risk scores. Overall, the patients in PARALLEL-HF were largely representative of contemporary ambulatory patients with HFrEF who are well treated with evidence-based therapies. PARALLEL-HF will determine whether sacubitril/valsartan provides similar improvements in clinical outcomes in Japanese HFrEF patients as observed in the PARADIGM-HF study.


Correlation of histopathologic findings with clinical outcome in necrotizing fasciitis.
Necrotizing fasciitis is an uncommon disease with high morbidity and mortality rates. Little is known about the role of histopathologic examination in disease prognosis. A retrospective study was conducted to determine what correlations, if any, exist between the histopathologic features of resected tissue in patients with necrotizing fasciitis and clinical outcome. Eighty-two cases of necrotizing fasciitis that occurred between January 1990 and December 2002 were identified. Histopathologic findings were available for review in 63 cases. A novel histopathologic classification scheme, based on hematoxylin-eosin and Gram stain results, was developed. The classification scheme included 3 stages: stage I, characterized by an intense neutrophilic response and an absence of bacteria in infected tissue; stage II, characterized by the presence of a moderate-to-severe neutrophilic response and positive Gram stain results or by minimal to absent neutrophilic response with a negative Gram stain result; and stage III, characterized by the presence of few or no polymorphonuclear leukocytes and a Gram stain result positive for bacteria during histopathologic examination. Patients with stage I findings had a significantly lower risk of death than patients with stage III findings (7.1% vs. 47%; odds ratio [OR], 0.1; 95% confidence interval [CI], 0.01-0.8; P=.03). Patients with stage II findings had a significantly lower mortality rate than patients with stage III findings (14.2% vs. 47%; OR, 0.2; 95% CI, 0.04-0.9; P=.04). Due to the small number of deaths (n=11) in patients for whom histopathologic examination of resected tissue was performed, multivariate analysis was not done. Results of this study suggest that histopathologic findings may correlate with clinical outcome in cases of necrotizing fasciitis. Because the histopathologic scheme is based on results of commonly available stains, it could be easily adopted for use in other institutions that could further evaluate its usefulness as a prognostic tool.