Postnatal development of fiber type composition in rabbit jaw and leg muscles.
We examined the difference in fiber type composition and cross-sectional areas during postnatal development in male rabbit jaw muscles and compared these with changes in leg muscles. The myosin heavy chain (MyHC) content of the fibers was determined by immunohistochemistry. No fiber type difference was found between the jaw muscles in 20-week-old rabbits. However, the way this adult fiber type composition was reached differed between the muscles. The deep temporalis, medial pterygoid, and superficial masseter displayed an increase in alpha fibers during early and a decrease during late postnatal development. Other jaw muscles displayed an increase in alpha fibers during early development only. In contrast, alpha fibers were not found in the soleus, in which fiber type changes were completed at week 4. The gastrocnemius muscle did not change its fiber type composition. Initially, fibers in jaw-opening muscles had larger cross-sectional areas than in other muscles, but they increased less during development. Although there were no large differences in the fiber type composition of muscles in young adult rabbits, large differences were found in the jaw muscles, but not in the leg muscles, during development. In part, these developmental changes in fiber percentages within the jaw muscles can be explained by functional modifications in this muscle group. In the present study, the deep temporalis, medial pterygoid, and superficial masseter showed the most dramatic percent changes in fibers during postnatal development.


Predictive Maintenance of Pins in the ECD Equipment for Cu Deposition in the Semiconductor Industry.
Nowadays, Predictive Maintenance is a mandatory tool to reduce the cost of production in the semiconductor industry. This paper considers as a case study a critical part of the electrochemical deposition system, namely, the four Pins that hold a wafer inside a chamber. The aim of the study is to replace the schedule of replacement of Pins presently based on fixed timing (Preventive Maintenance) with a Hardware/Software system that monitors the conditions of the Pins and signals possible conditions of failure (Predictive Maintenance). The system is composed of optical sensors endowed with an image processing methodology. The prototype built for this study includes one optical camera that simultaneously takes images of the four Pins on a roughly daily basis. Image processing includes a pre-processing phase where images taken by the camera at different times are coregistered and equalized to reduce variations in time due to movements of the system and to different lighting conditions. Then, some indicators are introduced based on statistical arguments that detect outlier conditions of each Pin. Such indicators are pixel-wise to identify small artifacts. Finally, criteria are indicated to distinguish artifacts due to normal operations in the chamber from issues prone to a failure of the Pin. An application (PINapp) with a user friendly interface has been developed that guides industry experts in monitoring the system and alerting in case of potential issues. The system has been validated on a plant at STMicroelctronics in Catania (Italy). The study allowed for understanding the mechanism that gives rise to the rupture of the Pins and to increase the time of replacement of the Pins by a factor at least 2, thus reducing downtime.


How we capitalised on casual PBL facilitators' expertise and experience to add value to our medical programme.
While problem-based learning (PBL) has been widely implemented in medical education, it has been acknowledged to be resource-intensive, particularly in terms of academics' time. In some institutions, such as Bond University (Australia), casual facilitators have been employed (paid hourly) to oversee the PBL tutorials. Apart from considerable experience as PBL facilitators, they also have expertise as allied health professionals or biomedical scientists. Several facilitators have educational qualifications. Recognising that their roles have expanded beyond the PBL tutorial room, we canvassed PBL facilitators in terms of their contributions to Bond University's medical programme. We can report that our facilitators have contributed to the renewal of Bond's medical programme, from design (e.g. curriculum structure, case-writing) to PBL case reviews. They are also involved in formative and summative assessment. Facilitators identified that, because of their prolonged involvement with the students in small groups, they consider themselves more than facilitators of student learning. They are role models and personal guides. Recognising the value these casually employed facilitators have added to our medical programme, we will continue to develop their skills in, for example, reviewing cases and assessment. We recommend that if institutions do employ casual facilitators, their expertise and experience can add value to the curriculum and to students' experience.


Iron and Folic Acid Consumption by the Ante-natal Mothers in a Rural Area of India in 2010.
The average rural Indian women enter her reproductive life, particularly in pregnancy, suffer from nutritional anemia due to iron deficiency. National program of India had implemented a strategy for supplementation of iron folic acid by means of iron folic acid (IFA) tablets at least 3 months during antenatal period. The study had been conducted to assess the proportion of pregnant mothers consumes the IFA tablets and the factors determine compliance. A cross-sectional, community-based study was conducted in rural area of India on 50 antenatal mothers by multistage sampling technique. The data were analyzed by using SPSS 16 (Statistical Package for the Social Sciences, ver. 16), Chicago, considering the level of significance at 95%. The IFA tablet was adequately consumed by 62% mother among the study population. The consumption is more among the mother who were explained properly than those who were not explained by the health worker (χ(2)= 4.529, P < 0.05). The compliance of iron folic acid tablets was still far behind to reach the National Goal though the service component are quite strong by the front line workers and health providers. An effort should be given at the level of front line health workers by training and re-training them to improve the compliance of IFA consumption.


Finding potentially new multimorbidity patterns of psychiatric and somatic diseases: exploring the use of literature-based discovery in primary care research.
Multimorbidity, the co-occurrence of two or more chronic medical conditions within a single individual, is increasingly becoming part of daily care of general medical practice. Literature-based discovery may help to investigate the patterns of multimorbidity and to integrate medical knowledge for improving healthcare delivery for individuals with co-occurring chronic conditions. To explore the usefulness of literature-based discovery in primary care research through the key-case of finding associations between psychiatric and somatic diseases relevant to general practice in a large biomedical literature database (Medline). By using literature based discovery for matching disease profiles as vectors in a high-dimensional associative concept space, co-occurrences of a broad spectrum of chronic medical conditions were matched for their potential in biomedicine. An experimental setting was chosen in parallel with expert evaluations and expert meetings to assess performance and to generate targets for integrating literature-based discovery in multidisciplinary medical research of psychiatric and somatic disease associations. Through stepwise reductions a reference set of 21,945 disease combinations was generated, from which a set of 166 combinations between psychiatric and somatic diseases was selected and assessed by text mining and expert evaluation. Literature-based discovery tools generate specific patterns of associations between psychiatric and somatic diseases: one subset was appraised as promising for further research; the other subset surprised the experts, leading to intricate discussions and further eliciting of frameworks of biomedical knowledge. These frameworks enable us to specify targets for further developing and integrating literature-based discovery in multidisciplinary research of general practice, psychology and psychiatry, and epidemiology.


Tuning corneal epithelial cell adhesive strength with varying crosslinker content in silicone hydrogel materials.
To quantify the effect of silicone hydrogel crosslink density on the adhesion at corneal epithelial cells/silicone hydrogel contact lens interface. A custom-built rheometer, referred to as the live cell monolayer rheometer, was used to measure the adhesive strengths between corneal epithelial cell monolayers and silicone hydrogel lens surfaces. The resulting stress relaxations of senofilcon A-derived silicone hydrogel materials with varying crosslinking densities and delefilcon A were tested. Senofilcon A-like materials labeled L1, L2, L3, L4, and L5 contained crosslinker concentrations of 1.2, 1.35, 1.5, 1.65, and 1.8 wt%, respectively. The residual modulus measured from the live cell monolayer rheometer provided a direct indication of adhesive attachment. Within the senofilcon-derived series, the adhesive strength shows a surprising minimum with respect to crosslink density. Specifically, L1 (1.20%) has the highest adhesive strength of 39.5 ± 11.2 Pa. The adhesive strength diminishes to a minimum of 11.2 ± 2.1 Pa for L3, whereafter it increases to 14.5 ± 2.5 Pa and 18.1 ± 5.1 Pa for L4 and L5, respectively. The delefilcon A lens exhibits a comparable adhesive strength of 27.8 ± 6.3 Pa to L1. These results demonstrated that increasing the crosslink density has a nonmonotonic influence on the adherence of lenses to mucin-expressing corneal epithelial cells, which suggests a competition mechanism at the cell/lens interface. Because the adhesiveness of contact lenses to ocular tissues may impact the comfort level for lens wearers and affect ease of removal, this study suggests that lens adhesion can be optimized through the control of crosslink density.


INA complex liaises the F1Fo-ATP synthase membrane motor modules.
The F1F0-ATP synthase translates a proton flux across the inner mitochondrial membrane into a mechanical rotation, driving anhydride bond formation in the catalytic portion. The complex's membrane-embedded motor forms a proteinaceous channel at the interface between Atp9 ring and Atp6. To prevent unrestricted proton flow dissipating the H+-gradient, channel formation is a critical and tightly controlled step during ATP synthase assembly. Here we show that the INA complex (INAC) acts at this decisive step promoting Atp9-ring association with Atp6. INAC binds to newly synthesized mitochondrial-encoded Atp6 and Atp8 in complex with maturation factors. INAC association is retained until the F1-portion is built on Atp6/8 and loss of INAC causes accumulation of the free F1. An independent complex is formed between INAC and the Atp9 ring. We conclude that INAC maintains assembly intermediates of the F1 F0-ATP synthase in a primed state for the terminal assembly step-motor module formation.


Structure of murine enterokinase (enteropeptidase) and expression in small intestine during development.
Enterokinase (enteropeptidase) is expressed only in proximal small intestine, where it initiates digestive enzyme activation by converting trypsinogen into trypsin. To investigate this restricted expression pattern, mouse enterokinase cDNA was cloned, and the distribution of enterokinase mRNA and enzymatic activity were determined in adult mice and during gestation. Analysis of enterokinase sequences showed that a mucinlike domain near the NH2 terminus is composed of repeated approximately 15-amino acid Ser/Thr-rich motifs. By Northern blotting and trypsinogen activation assays, enterokinase mRNA and enzymatic activity were undetectable in stomach, abundant in duodenum, and decreased distally until they were undetectable in midjejunum, ileum, and colon. By in situ mRNA hybridization, enterokinase mRNA was localized to the enterocytes throughout the villus. Expression was not observed in goblet cells, Paneth cells, or Brunner's glands. Enterokinase mRNA and enzymatic activity were not detected in the duodenum of fetal mice but were easily detected in the duodenum on postnatal days 2-6. Both enterokinase mRNA and enzymatic activity decreased to very low levels after day 7 but increased after weaning and reached a high level characteristic of adult life by day 60. Therefore, in mice, duodenal enterocytes are the major type of cells expressing enterokinase, which appears to be regulated at the level of mRNA abundance.


Emergence of SARS-CoV-2 Resistance with Monoclonal Antibody Therapy.
Resistance mutations to monoclonal antibody (mAb) therapy has been reported, but in the non-immunosuppressed population, it is unclear if in vivo emergence of SARS-CoV-2 resistance mutations alters either viral replication dynamics or therapeutic efficacy. In ACTIV-2/A5401, non-hospitalized participants with symptomatic SARS-CoV-2 infection were randomized to bamlanivimab (700mg or 7000mg) or placebo. Treatment-emergent resistance mutations were significantly more likely detected after bamlanivimab 700mg treatment than placebo (7% of 111 vs 0% of 112 participants, P=0.003). There were no treatment-emergent resistance mutations among the 48 participants who received bamlanivimab 7000mg. Participants with emerging mAb resistant virus had significantly higher pre-treatment nasopharyngeal and anterior nasal viral load. Intensive respiratory tract viral sampling revealed the dynamic nature of SARS-CoV-2 evolution, with evidence of rapid and sustained viral rebound after emergence of resistance mutations, and worsened symptom severity. Participants with emerging bamlanivimab resistance often accumulated additional polymorphisms found in current variants of concern/interest and associated with immune escape. These results highlight the potential for rapid emergence of resistance during mAb monotherapy treatment, resulting in prolonged high level respiratory tract viral loads and clinical worsening. Careful virologic assessment should be prioritized during the development and clinical implementation of antiviral treatments for COVID-19.


Methodological quality of studies and patient age as major sources of variation in efficacy estimates of influenza vaccination in healthy adults: a meta-analysis.
A meta-analysis of randomized and quasi-randomized control trials evaluating influenza vaccination efficacy in healthy adults was carried out to investigate potential sources of heterogeneity among the published studies. The pooled estimates of vaccine efficacy were 22% (95% CI = 16-28) for preventing clinically diagnosed cases of influenza and 63% (95% CI = 53-71) for preventing laboratory confirmed cases of influenza. These estimates are likely to vary substantially according to the age of the subjects, with vaccine efficacy decreasing significantly with the increase of the mean age of the patients, and to the methodological quality of trials, with low quality trials associated with an overestimation of influenza vaccine efficacy. Some evidence of publication bias was found, particularly when the outcome assessed was clinically diagnosed influenza. Further high-quality clinical trials, designed to facilitate future pooled analyses, are needed to provide definitive answers for policy-makers.


Interictal Epileptiform Discharge Detection Using Multi-Head Deep Convolutional Neural Network.
Interictal epileptiform discharges (IEDs) are intermittent electrophysiological events that occur in patients with epilepsy between seizures. Automated detection of IEDs helps clinician to identify cortical irritations and relations to seizure recurrence. It also reduces the necessity of visual inspection by physicians interpreting the EEG. This paper presents a novel deep learning-based approach that combines one-dimensional local binary pattern symbolization method with a regularized multi-head one-dimensional convolutional neural network to learn unique morphological patterns from different EEG sub-bands for IED detection. Experimentation using the Temple University Events corpus scalp EEG data shows promising performance, e.g. F1-score of 87.18%.


Pancreatin-induced liberation of starch/cyanidin 3-O-glucoside complexes from rice cooked with black soybean that exhibit slow hydrolysis.
Cyanidin 3-O-glucoside (C3G), which has various health-promoting functions, is contained in black soybean (BSB). In Japan and Korea, BSB is cooked with rice and the cooked rice appears purplish in colour. In this study, BSB was cooked with glutinous rice, non-glutinous rice, and high-amylose rice. The amount of C3G detected in high-amylose rice was greater than that detected in glutinous rice, suggesting that C3G combined more efficiently with amylose than with amylopectin. Pancreatin induced the liberation of starch/C3G complexes from the purplish cooked rice, and rate of the liberation was in the following order; glutinous rice < non-glutinous rice < high-amylose rice. The amylose/C3G complexes liberated from high-amylose rice was hydrolysed slowly, while the amylopectin/C3G complexes liberated from glutinous rice were hydrolysed into smaller amylopectin/C3G complexes that were difficult to further hydrolysis. Thus, C3G may be useful for preparing foods whose starch hydrolysis is slow.


In vivo genetic mutations define predominant functions of the human T-cell leukemia/lymphoma virus p12I protein.
The human T-cell leukemia/lymphoma virus type 1 (HTLV-1) ORF-I encodes a 99-amino acid hydrophobic membrane protein, p12(I), that affects receptors in different cellular compartments. We report here that proteolytic cleavage dictates different cellular localization and functions of p12(I). The removal of a noncanonical endoplasmic reticulum (ER) retention/retrieval signal within the amino terminus of p12(I) is necessary for trafficking to the Golgi apparatus and generation of a completely cleaved 8-kDa protein. The 8-kDa protein in turn traffics to the cell surface, is recruited to the immunologic synapse following T-cell receptor (TCR) ligation, and down-regulates TCR proximal signaling. The uncleaved 12-kDa form of p12(I) resides in the ER and interacts with the beta and gamma(c) chains of the interleukin-2 receptor (IL-2R), the heavy chain of the major histocompatibility complex (MHC) class I, as well as calreticulin and calnexin. Genetic analysis of ORF-I from ex vivo samples of HTLV-1-infected patients reveals predominant amino acid substitutions within ORF-I that affect proteolytic cleavage, suggesting that ER-associated functions of p12(I) may contribute to the survival and proliferation of the infected T cells in the host.


Bacterial isolates and drug susceptibility patterns of urinary tract infection among pregnant women at Muhimbili National Hospital in Tanzania.
Urinary tract infection (UTI) during pregnancy may cause complications such as pyelonephritis, hypertensive disease of pregnancy, anaemia, chronic renal failure, premature delivery and foetal mortality. This study aimed to identify the etiologic agents of UTI and to determine the patterns of antimicrobial drug susceptibility among pregnant women at Muhimbili National Hospital in Tanzania. Retrospective analysis of 200 mid-stream urine specimens processed for culture and antimicrobial drug susceptibility testing between January 2007 and December 2009 was carried out. Significant bacteriuria (> 105 colony forming units/mL of urine) was found in 42/200 (21%) specimens. Of the 42 isolates, the most commonly isolated bacteria were Escherichia coli 14 (33.3%), Klebsiella spp 9 (21.4%) coagulase negative Staphylococcus 7 (16.7%), Staphylococcus aureus 6 (14.3%), Proteus species 3 (7.1%) and Enterococcus species 3 (7.1%). Low rate to moderately high rate of antimicrobial drug resistance was observed against first line drugs namely, nitrofurantoin 18.7 % (n=16), co-trimoxazole 38.5 % (n=13) and ampicillin 57.7% (n=26). Relatively low rate of resistance was seen against second line drugs: ciprofloxacin 13.6 % (n=22) and amikacin 5 % (n=20). High rate of resistance was observed in third generation cephalosporin cefotaxime 31.2 % (n=16). Of the Gram-positive organisms tested against vancomicin and methicilin, resistance was found in 25 % (n=13) and 25 % (n=4), respectively. In conclusion, E coli was found to be the common cause of UTI among the pregnant women. Low to moderately high level of resistance was found in first line drugs while high level of resistance was found in third generation cephalosporin. It is recommended to monitor the levels of resistance for nitrofurantoin, fluoroquinolone and cefotaxime and to screen for Extended Spectrum Beta Lactamase production among cefotaxime resistant E. coli and Klebsiella spp.


Bacterial Flora of the Nose and Paranasal Sinuses Among Patients Over 65 Years Old with Chronic Rhinosinusitis Who Underwent Endoscopic Sinus Surgery.
Chronic rhinosinusitis (CRS) is one of the most common chronic diseases in the geriatric population. However, CRS inflammatory mechanisms in older people have not been thoroughly investigated. Our work aimed to analyze the bacterial flora of the nose and paranasal sinuses in patients with CRS over 65 years of age, including comorbidities, previously performed endoscopic sinus surgery (ESS), presence or absence of polyps and the extent of the inflammatory process. The study involved 529 patients between 18 and 84 years of age with chronic rhinosinusitis who underwent endoscopic sinus surgery. There were 101 patients separated over 65 years of age (M = 52, K = 49, mean age 69 ± 0.7 years). The control group consisted of 168 patients aged 18-40 years with CRS. The bacterial culture of material collected from the patients during ESS was analyzed. In the group of patients over 65 years of age, more frequent occurrence of Proteus spp. and Pseudomonas aeruginosa was found in comparison to younger patients. In older patients with bronchial asthma, the occurrence of S. aureus, Escherichia coli, and Citrobacter spp. was more frequent than in control group. Multiple sinus surgical procedures in older patients were associated with the dominance of Staphylococcus aureus and Escherichia coli, which was not demonstrated in the control group. There were no statistically significant differences between the occurrence of bacterial strain and the presence of polyps, both in the group of patients over 65 years of age as well as in the control group. The bacterial flora of patients with CRS is different in older and younger patients. A different therapeutic approach should be considered in older patients with CRS, but this problem requires further studies.


Longitudinal study of language therapy in 142 children and adolescents with autism spectrum disorders.
The last 25 years produced important changes in the Speech Language intervention for children with Autism Spectrum Disorders in the whole world. They refer to diagnostic criteria, research methods and systematization of data about diagnostic and intervention processes. In this context, the first results about the use of a recording system used on a specialized service highlights the importance of including information about each intervention process on reliable and accessible systems. This procedure allows the association among several types of data and studies with large populations, something that is not usual in the field.


Caring about risks. Are severely depressed patients competent to consent to research?
Depressed patients are often asked to take part in clinical research studies that carry risk. These patients are generally assumed to be mentally competent to consent to research, since depression often leaves a patient's cognitive abilities intact. In this article, it is argued that many severely depressed patients may not be competent to consent because they cannot be considered accountable for their decisions. The article presents 2 arguments: first, that it is unclear whether the decisions of some severely depressed patients are authentically theirs, and second, that some severely depressed patients may not have the appropriate minimal degree of concern for their own well-being. It is argued that assessments of competence must take account of emotional factors, and that, if severely depressed patients are incompetent to consent, research studies involving a poor risk-benefit ratio will be much more difficult to justify.


AKT signaling in regulating angiogenesis.
AKT is a central signaling molecule in regulating cell survival, proliferation, tumor growth and angiogenesis. Upstream components of AKT signaling pathway such as PI3K, PTEN, and Ras are commonly mutated in many human cancers. Recently it is found that AKT plays an important role in regulating normal vascularization and pathological angiogenesis. Angiogenesis is required for tumor growth and metastasis when tumor reaches more than 1 mm in diameter. This review focuses on the role and potential mechanism of AKT signaling in regulating angiogenesis. Recent studies have shown that AKT activation is necessary and sufficient to regulate VEGF and HIF-1 expression in human cancer cells. VEGF and HIF-1 are potent inducers of angiogenesis. It was found that AKT activation induces VEGF and HIF-1 expression through its two downstream molecules HDM2 and p70S6K1. On the other hand, AKT transmits the upstream signals from growth factors, cytokines, heavy metals, and oncogenes for regulating VEGF and HIF-1 expression in human cancer cells. AKT activation and VEGF expression can be inhibited by different natural compounds used for cancer prevention. Thus, inhibition of AKT and its downstream targets offers a new approach for targeting angiogenesis, which could be important for the development of new cancer therapeutics in the future.


Comparative study of the expression of DNA mismatch repair genes, the adenomatous polyposis coli gene and growth arrest DNA damage genes in melanoma recurrences and metastases.
The main goal of this study was to examine the expression of DNA mismatch repair genes (MLH1, MSH2, PMS1 and PMS2), the adenomatous polyposis coli (APC) gene and growth arrest DNA damage inducible (GADD) genes (GADD34, GADD45 and GADD153) in the different stages of melanoma recurrences and metastases, and to identify any mutual consistencies in their expression pattern. All the cases of primary melanoma examined showed a reduced expression of DNA repair genes. These results demonstrate that disturbances of DNA repair begin in the early stages of melanoma. No significant differences were found in the expression of these markers between cutaneous melanomas and their recurrences and metastases (P> 0.05). Eighteen significant correlations between markers were found in the primary melanomas, and 10 significant correlations were observed in the first recurrences of melanoma. In contrast, 27 statistically significant relationships were demonstrated in metastatic lymph nodes. The different correlations found in primary and metastatic tumours confirmed the hypothetical difference in marker interaction in the diagnostic groups investigated. Our results suggest that DNA repair genes may play an important role in the recurrence and metastasis of melanomas.


Mapping the signal-to-noise-ratios of cortical sources in magnetoencephalography and electroencephalography.
Although magnetoencephalography (MEG) and electroencephalography (EEG) have been available for decades, their relative merits are still debated. We examined regional differences in signal-to-noise-ratios (SNRs) of cortical sources in MEG and EEG. Data from four subjects were used to simulate focal and extended sources located on the cortical surface reconstructed from high-resolution magnetic resonance images. The SNR maps for MEG and EEG were found to be complementary. The SNR of deep sources was larger in EEG than in MEG, whereas the opposite was typically the case for superficial sources. Overall, the SNR maps were more uniform for EEG than for MEG. When using a noise model based on uniformly distributed random sources on the cortex, the SNR in MEG was found to be underestimated, compared with the maps obtained with noise estimated from actual recorded MEG and EEG data. With extended sources, the total area of cortex in which the SNR was higher in EEG than in MEG was larger than with focal sources. Clinically, SNR maps in a patient explained differential sensitivity of MEG and EEG in detecting epileptic activity. Our results emphasize the benefits of recording MEG and EEG simultaneously.


A novel retron that produces RNA-less msDNA in Escherichia coli using reverse transcriptase.
Bacterial retroelements, or retrons, use reverse transcriptase (RT) to produce a multicopy single-stranded DNA (msDNA) molecule that is covalently linked to RNA. In these studies we show that a retron from Escherichia coli 110, a clinical isolate, produces a novel RNA-less msDNA with a 5' phosphate residue. The msDNA is a 74-nucleotide single-stranded DNA molecule with a stable stem-loop structure without a mismatched base pair. Only the genes encoding msDNA (msd), msdRNA (msr), and RT (ret) are required to produce the msDNA molecule. The organization of these genes on the retron was similar to that of other elements producing branched msDNA-RNA. The conserved guanine, which is the branched residue in msDNA-RNA complexes and is essential for branch formation, is also present. Site-directed mutagenesis showed that this guanine is essential for the production of RNA-less msDNA. We postulate that the RNA-less msDNA in strain 110 is produced by nucleolytic cleavage of the branched msDNA-RNA compound.


Compulsory drug detention centers in China, Cambodia, Vietnam, and Laos: health and human rights abuses.
According to official accounts, in 2012 more than 235,000 people were detained in over 1,000 compulsory drug detention centers in East and Southeast Asia. Between July 2007 and May 2013, in-depth interviews were conducted with 195 individuals recently released from drug detention centers in China, Cambodia, Vietnam, and Laos. Individuals reported being held for up to five years in drug detention centers without clinical determination of drug dependency or due process, and being denied evidence-based drug treatment as well as other basic health services. Many individuals reported being forced to perform arduous physical exercise or military-style drills. Forced labor was reported by all individuals having been detained in Vietnam, and some held in Cambodia and China. Physical—and less often, sexual—abuse was reported among those held in each country. Long-term, compulsory detention for treatment of drug dependency is counter to established principles of medical care and violates a wide range of human rights, including the right to health. Individuals held in drug detention centers in China, Cambodia, Vietnam, and Laos are subject to torture and cruel, inhuman, and degrading treatment.


A Helminth Protease Inhibitor Modulates the Lipopolysaccharide-Induced Proinflammatory Phenotype of Microglia in vitro.
The aim of this study was to examine whether the natural protease inhibitor Av-cystatin (rAv17) of the parasitic nematode Acanthocheilonema viteae exerts anti-inflammatory effects in an in vitro model of lipopolysaccharide (LPS)-activated microglia. Primary microglia were harvested from the brains of 2-day-old Wistar rats and cultured with or without rAv17 (250 nM). After 6 and 24 h the release of nitric oxide (Griess reagent) and TNF-α (ELISA) was measured in the supernatant. Real-time PCR was performed after 2, 6 and 24 h of culture to measure the mRNA expression of IL-1β, IL-6, TNF-α, COX-2, iNOS and IL-10. To address the involved signaling pathways, nuclear NF-x0138;B translocation was visualized by immunocytochemistry. Morphological changes of microglia were analyzed by Coomassie blue staining. Differences between groups were calculated using one-way ANOVA with Bonferroni's post hoc test. Morphological analysis indicated that LPS-induced microglial transformation towards an amoeboid morphology is inhibited by rAv17. Av-cystatin caused a time-dependent downregulation of proinflammatory cytokines, iNOS and COX-2 mRNA expression, respectively. This was paralleled by an upregulated expression of IL-10 in resting as well as in LPS-stimulated microglia. Av-cystatin reduced the release of NO and TNF-α in the culture supernatant. Immunocytochemical staining demonstrated an attenuated translocation of NF-x0138;B by Av-cystatin in response to LPS. In addition, Western blot analysis revealed a rAv17-dependent reduction of the LPS-induced ERK1/2-pathway activation. The parasite-derived secretion product Av-cystatin inhibits proinflammatory mechanisms of LPS-induced microglia with IL-10, a potential key mediator.


Locus-Specific Regulation of Xist Expression Using the CRISPR-Cas9-Based System.
DNA methylation inhibitor or loss and gain of function of DNA methylation key players were widely used to investigate the regulation of X inactive-specific transcript (Xist) expression by DNA methylation, which results in global change of DNA methylation. Here, we reported a novel method for regulation of Xist using the widely used clustered regularly interspaced short palindromic repeat (CRISPR)-Cas system. First, Xist expression was increased in 5-aza-2'-deoxycytidine-treated female goat fibroblast cells. Second, three single-guide RNAs (sgRNAs) that target the Xist differential methylation region (DMR) were inserted to deactivated Cas9 (dCas9) nuclease and the catalytic domain of the DNA methyltransferase Dnmt3a coexpression plasmid. Bisulfite PCR analysis and quantitative real-time PCR revealed that the methylation level of the DMR was significantly increased, while the expression of Xist was downregulated in all three sgRNAs, compared with the mock-transfected cells. Third, the methylation activity at the sites of 37 bp from the protospacer-adjacent motif sequence showed the strong change relative to the mock-transfected cells. Furthermore, genome-wide DNA methylation and expression of the DNA methylation key players were not statistically changed in all three sgRNAs. Therefore, we confirmed that Xist expression was regulated by DNA methylation, and directed DNA methylation of Xist DMR at locus-specific solution decreased Xist expression.


Mutations in a carboxy-terminal region of vesicular stomatitis virus glycoprotein G that affect membrane fusion activity.
The envelope glycoprotein G of vesicular stomatitis virus induces membrane fusion at acidic pH. A highly conserved amino terminal region spanning residues 123 to 137 has previously been identified as an internal fusion domain. Here we have substituted specific amino acids within a carboxy terminal region, conserved in five vesiculoviruses encompassing residues 395 to 418, and studied the effect of these mutations on membrane fusion at acid pH and pH-dependent conformational change. Substitution of conserved Gly 395, Gly 404, Gly 406, Asp 409, and Asp 411 with Glu, Ala, Ala, Asn, and Asn, respectively, decreased the cell-cell fusion efficiency, as well as reduced the pH threshold of membrane fusion. Mutation of Gly 404 and Asp 409 to Lys and Ala, respectively, abolished the fusion activity. Mutant Gly 404 Lys also showed markedly altered resistance to trypsin digestion at acidic pH. These results suggest that the region between amino acids 395 to 418 is important for the fusogenic activity of the G protein. The possible role of this domain in conformational changes involved in fusion activity of VSV G is also discussed.


Nuclear Ca2+ and CaM kinase IV specify hormonal- and Notch-responsiveness.
Many neuronal processes require gene activation by synaptically evoked Ca(2+) transients. Ca(2+)-dependent signal pathways activate some transcription factors outright, but here we report that such signals also potentiate the activation of nuclear receptors by their cognate hormone, and of CBF1 by Notch, transcription factors hitherto not thought to be Ca(2+)-responsive. This potentiation is occluded by histone deacetylase inhibition, indicating a mechanism involving inactivation of co-repressors associated with these transcription factors. Synaptic activity, acting via the nuclear Ca(2+)-dependent activation of CaM kinase IV, triggers the disruption of subnuclear domains containing class II histone deacetylases (HDACs) and silencing mediator of retinoic acid and thyroid hormone receptors (SMRT), a broad-specificity co-repressor which represses nuclear hormone receptors and CBF1. The sequential loss of class II HDACs and SMRT from the subnuclear domains, followed by nuclear export, is associated with disruption of SMRT interaction with its target transcription factors and sensitization of these factors to their activating signal. Counterbalancing these changes, protein phosphatase 1 promotes nuclear localization of SMRT and inactivation of nuclear receptors and CBF1. Thus, the synaptically controlled kinase-phosphatase balance of the neuron determines the efficacy of SMRT-mediated repression and the signal-responsiveness of a variety of transcription factors.


Antihyperalgesic effects of delta opioid agonists in a rat model of chronic inflammation.
Opioid receptors in the brain activate descending pain pathways to inhibit the nociceptive response to acute noxious stimuli. The aim of the present study was to clarify the role of supraspinal opioid receptors in modulating the nociceptive response to persistent inflammation in rats. Subcutaneous administration of 50 microl of complete Freund's Adjuvant (CFA) into the plantar surface of the hindpaw induced a significant decrease in paw withdrawal latency to thermal stimuli (P<0.01) at 24 h post-injection. Intracerebroventricular (i.c.v.) administration of the mu opioid receptor agonists, DAMGO and morphine, and the delta opioid receptor agonists, deltorphin II and SNC80, significantly reversed the hyperalgesic response associated with peripheral inflammation in a dose-dependent manner (P<0.0001). The mu and delta agonists also significantly attenuated the antinociceptive response to acute thermal stimulation in rats (P<0.001). However, deltorphin II and SNC80 were less potent, and in the case of SNC80 less efficacious, in modulating the response to acute thermal nociception in comparison to hyperalgesia associated with persistent inflammation. These results indicate that mu and delta opioid receptors in the brain modulate descending pain pathways to attenuate the nociceptive response to acute thermal stimuli in both normal and inflamed tissues. The heightened response to delta agonists in the hyperalgesia model suggests that delta opioid receptors in the brain are promising targets for the treatment of pain arising from chronic inflammation.


Generation and Propagation of MALDI Ion Packets Probed by Sheet-Like Nanosecond UV Laser Light.
A sheet-like ultraviolet (UV) probe laser is used to investigate the ejection and propagation of ion packets of matrix CHCA, which are produced by matrix-assisted laser desorption and ionization (MALDI). Laser irradiation of the expanding MALDI plume induced photodissociation of the CHCA-related ions, which existed in a sheet-like volume, leading to their absence in their MALDI signal profiles. The MALDI spectra were measured under varying conditions: the temporal delay of the lasers and the distance of the sheet-like probe laser from the MALDI sample surface. It was found that the center of the (CHCA)H+ packets were ejected at 46±11 ns after MALDI laser irradiation, while the (CHCA)2H+ packets were ejected at 64±12 ns, regardless of the magnitude of acceleration static high-voltage in 3.5-5.5 kV. This suggests that (CHCA)2H+ is formed by a proton transfer reaction from (CHCA)H+ to (CHCA)2 in the heated condensed phase and/or near the surface. This study represents the first experimental determination of ion ejection time in the MALDI process, which is also applicable to other species in the MALDI plume.


Maternal Race/Ethnicity and Postpartum Diabetes Screening: A Systematic Review and Meta-Analysis.
Introduction: Fifty percent of women with gestational diabetes mellitus (GDM) may progress to type 2 diabetes with highest risk among black women. This study aims to characterize postpartum diabetes screening rates among U.S. women with GDM by racial and ethnic group to characterize potential disparities. Materials and Methods: A standardized search of Ovid-Medline, Embase, Scopus, Cumulative Index of Nursing and Allied Health Literature (CINAHL), Cochrane, ProQuest, and Clinicaltrials.gov was conducted through October 12, 2018. Of 1,555 titles reviewed, 27 studies met inclusion criteria. Meta-proportion routines with random-effects models estimated pooled postpartum screening proportion effect size (ES) with 95% confidence interval (CI) by racial and ethnic group. Heterogeneity was measured using Cochrane's Q and Higgins I2 tests. Data were stratified by intervention and data source. Results: There were 96,439 women, of whom 81,930 had race/ethnicity recorded. Heterogeneity was high (I2 = 99.7%). Postpartum screening rates were low (pooled ES 42% [95% CI 35%-48%]). Point estimates for pooled screening proportions were lower among white (pooled ES 35% [95% CI 28%-42%]) and black (pooled ES 33% [95% CI 24%-42%]) women than among Hispanic (pooled ES 45% [95% CI 37%-53%]) and Asian (pooled ES 50% [95% CI 41%-58%]) women. Interventions to improve screening were most common and effective among Hispanic women. Discussion: Postpartum screening for diabetes after GDM remains low, and black women have among the lowest postpartum screening rates despite highest risk for type 2 diabetes progression. Reporting of race/ethnicity, screening methods, and screening time frames varied across studies. Conclusion: Future studies must standardize racial/ethnic data reporting and examine interventions that address postpartum diabetes screening and prevention.


[Focus of childhood obesity from pediatrics].
The prevalences of overweight and obesity have increased dramatically in the last two decades in the adult and children population. The Organization for Cooperation and Economic Development reported in 2010 that Mexico ranks first worldwide in childhood obesity. The 2006 National Health and Nutrition Survey reported that one of every three teenagers are overweight and obese. In the last decades, pediatric hospitals in different parts of the world reported the prevalence of secondary malnutrition, since in those days overweight and obesity did not represent health problems. Currently, the prevalence of overweight and obesity has been scarcely studied in pediatric hospitals. In the Hospital de Pediatría (Children's Hospital) of the Instituto Mexicano del Seguro Social's Centro Médico Nacional de Occidente it is reported a prevalence of overweight of 15.4 % and obesity of 12.2 %, which reflects a nutritional transition.Due to the high prevalence of overweight and obesity in this pediatric hospital of reference, one could conclude that the pediatrician should be able to make a correct evaluation of the nutritional state, because, if he does not detect these problems, we will be condemning children to suffer from a chronic disease for the rest of their lives, and with all the implications in the short, medium and long term.


Effect of regulated vitamin D increase on vascular markers in patients with chronic kidney disease: A systematic review and meta-analysis of randomized controlled trials.
The effect of increased vitamin D levels on vascular function in patients with chronic kidney disease (CKD) is controversial. This meta-analysis aimed to assess the effect of regulated vitamin D increase on vascular markers in patients with CKD. We searched PubMed, Web of Science, Embase and ClinicalTrials.gov from database inception up until July 21, 2023. We included randomized controlled trials assessing the effects of using vitamin D and its analogues on vascular function in patients with CKD. Fixed-effects and random-effects model analyses were performed using weighted mean difference effects for each trial by heterogeneity (I2) assessment. Primary outcomes encompassed blood flow-mediated dilation (FMD)、pulse wave velocity (PWV) and augmentation index (AIx). From 1964 records we selected 12 trials, 5 (n = 331) on FMD, 8 (n = 626) on PWV and 4 (n = 393) on AIx. Vitamin D and VDRA supplementation failed to significantly improve FMD (WMD 1.68%; 95% CI -0.18 to 3.53; P = 0.08; I2 = 88%)、PWV (WMD -0.41 m/s; 95%CI -0.95 to 0.13; P = 0.14; I2 = 57%)and AIx (WMD -0.53%; 95%CI -1.69 to 0.63; P = 0.37; I2 = 0%). Subgroup analysis revealed that 2 μg paricalcitol significantly improved FMD (WMD 2.09%; 95%CI 1.28 to 2.90; P < 0.00001); I2 = 0%), as did cholecalciferol (WMD 5.49%; 95% CI 4.35 to 6.63; P < 0.00001). Supplementation vitamin D and VDRA are associated with improved vascular function as measured by FMD, but not arterial stiffness as measured by PWV and AIx, tentatively suggesting that regulating the increase of vitamin D could not potentially reduce the incidence of cardiovascular disease.


Recovery of upper gastrointestinal bowel movement after rectosigmoid cancer surgery: a pilot transit analysis.
Postoperative gastrointestinal bowel transit right after colorectal resection has not yet been clarified. Thirty patients with rectosigmoid cancer were treated in this pilot study. The nasogastric tube was removed on the first postoperative day. One Sitzmarks capsule was given to each patient on the second postoperative day. Abdominal X-rays were taken at 3, 6, 8, 24, 48, and 72 hours after capsule intake. Distribution of the remaining Sitzmarks capsules were counted on X-ray films to clarify postoperative gastrointestinal movement after bowel resection. All Sitzmarks capsules were observed in the stomach at 3 and 6 hours after capsule intake. At 8 hours (second postoperative day), the Sitzmarks capsules were distributed from the stomach to the small intestine. Sitzmarks capsules were distributed in the right side colon at 24 hours (third postoperative day) after intake. Although the main distribution was still in the right side colon, several patients had evacuations accompanied by the disappearance of the Sitzmarks capsules. In 50% of the patients, it took approximately 72 hours (fifth postoperative day) for the first defecation after intake of the capsules. However, the Sitzmarks capsules remained mainly in the right side colon. Eight hours after intake, the majority of the Sitzmarks capsules shifted to the small intestine. Therefore, medication or feeding should be safely possible starting on the second postoperative day. There was no particular impact of bowel resection on upper gastrointestinal transit in patients with rectosigmoid cancer.


Interferon-gamma enhances radiation-induced cell death via downregulation of Chk1.
Interferon-gamma (IFNγ) is a cytokine with roles in immune responses as well as in tumor control. Interferon is often used in cancer treatment together with other therapies. Here we report a novel approach to enhancement of cancer cell killing by combined treatment of IFNγ with ionizing radiation. We found that IFNγ treatment alone in HeLa cells induced phosphorylation of Chk1 in a time- and dose-dependent manner, and resulted in cell arrest. Moreover IFNγ treatment was correlated with attenuation of Chk1 as the treatment shortened protein half-life of Chk1. As Chk1 is an essential cell cycle regulator for viability after DNA damage, attenuation of Chk1 by IFNγ pre-treatment in HeLa cells resulted in increased cell death following ionizing radiation about 2-folds than ionizing radiation treatment alone whereas IFNγ treatment alone had little effect on cell death. X-linked inhibitor of apoptosis-associated factor 1 (XAF1), an IFN-induced gene, seems to partly regulate IFNγ-induced Chk1 destabilization and radiation sensitivity because transient depletion of XAF1 by siRNA prevented IFNγ-induced Chk1 attenuation and partly protected cells from IFNγ-enhanced radiation cell killing. Therefore the results provide a novel rationale to combine IFNγ pretreatment and DNA-damaging anti-cancer drugs such as ionizing radiation to enhance cancer cell killing.


Repertoire requirements of CD4+ T cells that prevent spontaneous autoimmune encephalomyelitis.
Spontaneous experimental autoimmune encephalomyelitis arises in 100% of mice exclusively harboring myelin basic protein-specific T cells, and can be prevented by a single injection of CD4+ T cells obtained from normal donors. Given the powerful regulatory effect of the transferred T cells, we further investigated their properties, and, in particular, their repertoire requirements. Transfer of monoclonal OVA-specific CD4+ T cells did not confer protection from disease even when present at very high proportions (about 80% of total lymphocytes). Lack of protection was also evident after immunization of these animals with OVA, indicating that not just any postthymic CD4+ T cells has the potential to become regulatory. However, protection was conferred by cells bearing limited TCR diversity, including cells expressing a single Valpha4 TCR chain or cells lacking N nucleotides. We also investigated whether coexpression of the myelin basic protein-specific TCR with another TCR in a single cell would alter either pathogenesis or regulation. This was not the case, as myelin basic protein-specific/OVA-specific recombinase activating gene-1-/- double TCR transgenic mice still developed experimental autoimmune encephalomyelitis spontaneously even after immunization with OVA. Based on this evidence, we conclude that CD4+ T regulatory cells do not express canonical TCRs and that the altered signaling properties brought about by coexpression of two TCRs are not sufficient for the generation of regulatory T cells. Instead, our results indicate that regulatory T cells belong to a population displaying wide TCR diversity, but in which TCR specificity is central to their protective function.


Illumination estimation from specular highlight in a multi-spectral image.
The reflection spectrum of an object characterizes its surface material, but for non-Lambertian scenes, the recorded spectrum often deviates owing to specular contamination. To compensate for this deviation, the illumination spectrum is required, and it can be estimated from specularity. However, existing illumination-estimation methods often degenerate in challenging cases, especially when only weak specularity exists. By adopting the dichromatic reflection model, which formulates a specular-influenced image as a linear combination of diffuse and specular components, this paper explores two individual priors and one mutual prior upon these two components: (i) The chromaticity of a specular component is identical over all the pixels. (ii) The diffuse component of a specular-contaminated pixel can be reconstructed using its specular-free counterpart describing the same material. (iii) The spectrum of illumination usually has low correlation with that of diffuse reflection. A general optimization framework is proposed to estimate the illumination spectrum from the specular component robustly and accurately. The results of both simulation and real experiments demonstrate the robustness and accuracy of our method.


Differential glucocorticoid responsiveness of dialysis patients' lymphocytes.
To evaluate in vitro glucocorticoid responsiveness of phytohemagglutinin (PHA)-stimulated lymphocytes from peritoneal dialysis (PD) patients compared to hemodialysis (HD) patients. Cross-sectional study of prevalent PD and HD patients and concurrent control subjects. Urban outpatient dialysis unit. 20 HD, 14 PD, and 20 control subjects. Using standard lymphocyte culture techniques, the concentration of prednisolone (P) and methylprednisolone (MP) required to cause 50% inhibition (IC50) of the proliferative response to phytohemagglutinin (PHA) was determined from dose-response curves. There was considerable heterogeneity in the sensitivities of individual patients' PBMC to glucocorticoid inhibition, especially those of HD patients' cells to P. The mean +/- SD IC50 for MP was significantly (p < or = 0.001) lower than that for P in each cohort: PD 11 +/- 5 vs. 34 +/- 18 ng/mL; HD 22 +/- 14 vs. 89 +/- 43 ng/mL; control subjects 14 +/- 11 vs. 55 +/- 56 ng/mL. Interestingly, the IC50 for both P and MP was significantly higher in HD than in either PD or controls (ANOVA, P: F = 6.56, p = 0.003; MP: F = 3.77, p = 0.03), indicating decreased sensitivity of HD lymphocytes to both drugs. There were no significant differences in mean IC50 values for either P or MP between PD and controls. No correlations were found between IC50 for either P or MP and patient age, gender, duration of dialysis, serum creatinine, serum albumin, or parathyroid hormone level. In vitro glucocorticoid responsiveness of dialysis patients' lymphocytes appears to be influenced by dialysis modality, but the factor(s) involved remains to be determined. The greater sensitivity of PD lymphocytes to both P and MP might result in better immunosuppression and less severe rejection after renal transplantation. MP may be particularly advantageous following renal transplantation for any patient manifesting relative or absolute in vitro resistance to P.


Legal landmark for community care of former psychiatric patients.
A recent judgment of the Lands Tribunal has set an important precedent which should decrease the potential for restrictive covenants to thwart the development of the government's care in the community programme. In 1989 a Worthing couple were permitted to convert their own house into a residential care home for former psychiatric patients. Judge B Marder, QC, ruled on 12 January 1993 that the "public interest" outweighed a contractual stipulation that the property should be used only for residential, non-business purposes. This is a radical change in the Lands Tribunal's view of mental illness.


Murine genes related to the Drosophila AbdB homeotic genes are sequentially expressed during development of the posterior part of the body.
The cloning, characterization and developmental expression patterns of two novel murine Hox genes, Hox-4.6 and Hox-4.7, are reported. Structural data allow us to classify the four Hox-4 genes located in the most upstream (5') position in the HOX-4 complex as members of a large family of homeogenes related to the Drosophila homeotic gene Abdominal B (AbdB). It therefore appears that these vertebrate genes are derived from a selective amplification of an ancestral gene which gave rise, during evolution, to the most posterior of the insect homeotic genes so far described. In agreement with the structural colinearity, these genes have very posteriorly restricted expression profiles. In addition, their developmental expression is temporally regulated according to a cranio-caudal sequence which parallels the physical ordering of these genes along the chromosome. We discuss the phylogenetic alternative in the evolution of genetic complexity by amplifying either genes or regulatory sequences, as exemplified by this system in the mouse and Drosophila. Furthermore, the possible role of 'temporal colinearity' in the ontogeny of all coelomic (metamerized) metazoans showing a temporal anteroposterior morphogenetic progression is addressed.


Drop foot after knee dislocation: evaluation and treatment.
Although knee dislocations are relatively rare, serious complications make treatment difficult. Common peroneal nerve (CPN) palsy is a debilitating complication and its incidence has been reported as high as 50%. Even after successful ligament construction, unresolved CPN palsy is a major factor contributing to poor outcomes after knee dislocations. CPN palsy is more common with open dislocations, rotatory dislocations, and especially occurs in patients with posterolateral corner injuries. CPN palsy can be readily diagnosed clinically, although a high index of suspicion is needed. The risk versus benefits of surgical exploration in the acute setting is still under debate. Conservative management can be appropriate in the early phase of treatment, however, for persistent nerve damage, surgery is the treatment of choice because it results in better functional outcomes. Neurolysis, primary nerve repair, nerve grafting, and posterior tibialis tendon transfer have all been used by surgeons as viable surgical treatment options. As late surgical treatment of CPN typically results in poor prognosis, awareness of this injury, thorough physical examination and documentation of the nerve injury, and close follow-up are of paramount importance.


BreastPRS is a gene expression assay that stratifies intermediate-risk Oncotype DX patients into high- or low-risk for disease recurrence.
Molecular prognostic assays, such as Oncotype DX, are increasingly incorporated into the management of patients with invasive breast carcinoma. BreastPRS is a new molecular assay developed and validated from a meta-analysis of publically available genomic datasets. We applied the assay to matched fresh-frozen (FF) and formalin-fixed paraffin-embedded (FFPE) tumor samples to translate the assay to FFPE. A linear relationship of the BreastPRS prognostic score was observed between tissue preservation formats. BreastPRS recurrence scores were compared with Oncotype DX recurrence scores from 246 patients with invasive breast carcinoma and known Oncotype DX results. Using this series, a 120-gene Oncotype DX approximation algorithm was trained to predict Oncotype DX risk groups and then applied to series of untreated, node-negative, estrogen receptor (ER)-positive patients from previously published studies with known clinical outcomes. Correlation of recurrence score and risk group between Oncotype DX and BreastPRS was statistically significant (P < 0.0001). 59 of 260 (23 %) patients from four previously published studies were classified as intermediate-risk when the 120-gene Oncotype DX approximation algorithm was applied. BreastPRS reclassified the 59 patients into binary risk groups (high- vs. low-risk). 23 (39 %) patients were classified as low-risk and 36 (61 %) as high-risk (P = 0.029, HR: 3.64, 95 % CI: 1.40-9.50). At 10 years from diagnosis, the low-risk group had a 90 % recurrence-free survival (RFS) rate compared to 60 % for the high-risk group. BreastPRS recurrence score is comparable with Oncotype DX and can reclassify Oncotype DX intermediate-risk patients into two groups with significant differences in RFS. Further studies are needed to validate these findings.


[Signal-to-noise ratio measurement in parallel MRI with subtraction mapping and consecutive methods].
When measuring the signal-to-noise ratio (SNR) of an image the used parallel magnetic resonance imaging, it was confirmed that there was a problem in the application of past SNR measurement. With the method of measuring the noise from the background signal, SNR with parallel imaging was higher than that without parallel imaging. In the subtraction method (NEMA standard), which sets a wide region of interest, the white noise was not evaluated correctly although SNR was close to the theoretical value. We proposed two techniques because SNR in parallel imaging was not uniform according to inhomogeneity of the coil sensitivity distribution and geometry factor. Using the first method (subtraction mapping), two images were scanned with identical parameters. The SNR in each pixel divided the running mean (7 by 7 pixels in neighborhood) by standard deviation/radical2 in the same region of interest. Using the second (consecutive) method, more than fifty consecutive scans of the uniform phantom were obtained with identical scan parameters. Then the SNR was calculated from the ratio of mean signal intensity to the standard deviation in each pixel on a series of images. Moreover, geometry factors were calculated from SNRs with and without parallel imaging. The SNR and geometry factor using parallel imaging in the subtraction mapping method agreed with those of the consecutive method. Both methods make it possible to obtain a more detailed determination of SNR in parallel imaging and to calculate the geometry factor.


Update on hemolytic uremic syndrome: Diagnostic and therapeutic recommendations.
Hemolytic uremic syndrome (HUS) is a rare disease. In this work the authors review the recent findings on HUS, considering the different etiologic and pathogenetic classifications. New findings in genetics and, in particular, mutations of genes that encode the complement-regulatory proteins have improved our understanding of atypical HUS. Similarly, the complement proteins are clearly involved in all types of thrombotic microangiopathy: typical HUS, atypical HUS and thrombotic thrombocytopenic purpura (TTP). Furthermore, several secondary HUS appear to be related to abnormalities in complement genes in predisposed patients. The authors highlight the therapeutic aspects of this rare disease, examining both "traditional therapy" (including plasma therapy, kidney and kidney-liver transplantation) and "new therapies". The latter include anti-Shiga-toxin antibodies and anti-C5 monoclonal antibody "eculizumab". Eculizumab has been recently launched for the treatment of the atypical HUS, but it appears to be effective in the treatment of typical HUS and in TTP. Future therapies are in phases I and II. They include anti-C5 antibodies, which are more purified, less immunogenic and absorbed orally and, anti-C3 antibodies, which are more powerful, but potentially less safe. Additionally, infusions of recombinant complement-regulatory proteins are a potential future therapy.


Regulation of tissue repair and regeneration by electric fields.
Endogenous electric fields (EFs) have been detected at wounds and damaged tissues. The potential roles of EFs in tissue repair and regeneration have been an intriguing topic for centuries. Recent researches have provided significant insights into how naturally occurring EFs may participate in the control of tissue repair and regeneration. Applied EFs equivalent to the size of fields measured in vivo direct cell migration, cell proliferation and nerve sprouting at wounds. More remarkably, physiological EFs are a guidance cue that directs cell migration which overrides other well accepted directional signals including initial injury stimulation, wound void, contact inhibition release, population pressure and chemotaxis. EFs activate many intracellular signaling pathways in a directional manner. Modulation of endogenous wound EFs affects epithelial cell migration, cell proliferation, and nerve growth at cornea wounds in vivo. Electric stimulation is being tested clinically for the treatments of bone fracture, wound healing and spinal cord injury. EFs thus may represent a novel type of signaling paradigm in tissue repair and regeneration. Combination of the electric stimulation and other well understood biochemical regulatory mechanisms may offer powerful and effective therapies for tissue repair and regeneration. This review introduces experimental evidence for the existence of endogenous EFs and discusses their roles in tissue repair and regeneration.


Ballistic dynamics of Airy beams.
We demonstrate both theoretically and experimentally that optical Airy beams propagating in free space can perform ballistic dynamics akin to those of projectiles moving under the action of gravity. The parabolic trajectories of these beams as well as the motion of their center of gravity were observed in good agreement with theory. The possibility of circumventing an obstacle placed in the path of the Airy beam is discussed.


MYC rearrangement and translocations involving band 8q24 in diffuse large cell lymphomas.
The configuration of the MYC gene in diffuse large cell lymphomas (DLCL) with translocations involving band 8q24 [t(8q24)] has not been systematically studied. We collected cytogenetic and clinical data on 171 consecutive cases of DLCL, including cleaved, noncleaved, and immunoblastic types, of which 96 had DNA available and 124 had abnormal karyotypes. The cases with DNA available were evaluated for MYC rearrangement (MYC-R) by Southern hybridization of EcoRI-digested tumor DNA using an exon-1 probe, a combination of probe and enzyme known to detect over 85% of breaks in sporadic Burkitt's lymphoma. In cases studied at diagnosis, MYC-R, t(8;14)(q24;q32), or other t(8q24) were not prognostically significant. Among the 124 cases with karyotypic abnormalities, seropositivity for human immunodeficiency virus was significantly more common in cases with a t(8q24) (72%) than in cases without it (9%) (P less than .05). Of the four cases with an MYC -R, two had a t(8;14), one had a t(7;8;14)(p15;q24;q32), and one had a t(8;?)(q24;?) and a del(8)(q24). In the three previous cases with translocations involving 8q24 and 14q32, comigration of the rearranged MYC band with either the J region or the switch-mu region of the Ig heavy chain gene could not be demonstrated, leaving the 14q32 breakpoint undefined at the molecular level. Among the remaining 72 cases where both an abnormal karyotype and molecular data were available, 11 had a t(8q24), either t(8;14) or t(8;22)(q24;q11), in the absence of an MYC-R. In these cases, the 8q24 break was presumably located outside of the EcoRI MYC fragment. All 15 cases with a t(8q24) were also screened for point mutations in the PvuII site in the first exon of MYC; two cases that were not MYC-R showed loss of this restriction site. These results indicate that in most DLCL with t(8;14) or other t(8q24), the 8q24 breakpoint lies away from the MYC gene; in a minority of these cases, point mutations in regulatory noncoding regions were detected.


Optimization and parallelization of the discrete ordinate method for radiation transport simulation in OpenFOAM: Hierarchical combination of shared and distributed memory approaches.
This paper describes the reduction in memory and computational time for the simulation of complex radiation transport problems with the discrete ordinate method (DOM) model in the open-source computational fluid dynamics platform OpenFOAM. Finite volume models require storage of vector variables in each spatial cell; DOM introduces two additional discretizations, in direction and wavelength, making memory a limiting factor. Using specific classes for radiation sources data, changing the store of fluxes and other minor changes allowed a reduction of 75% in memory requirements. Besides, a hierarchical parallelization was developed, where each node of the standard parallelization uses several computing threads, allowing higher speed and scalability of the problem. This architecture, combined with optimization of some parts of the code, allowed a global speedup of x15. This relevant reduction in time and memory of radiation transport opens a new horizon of applications previously unaffordable.


Liver Angiopoietin-2 Is a Key Predictor of De Novo or Recurrent Hepatocellular Cancer After Hepatitis C Virus Direct-Acting Antivirals.
Recent reports suggested that direct acting antivirals (DAAs) might favor hepatocellular carcinoma (HCC). In study 1, we studied the proangiogenic liver microenvironment in 242 DAA-treated chronic hepatitis C patients with advanced fibrosis. Angiopoietin-2 (ANGPT2) expression was studied in tissue (cirrhotic and/or neoplastic) from recurrent, de novo, nonrecurrent HCC, or patients never developing HCC. Circulating ANGPT2,vascular endothelial growth factor (VEGF), and C-reactive protein (CRP) were also measured. In study 2, we searched for factors associated with de novo HCC in 257 patients with cirrhosis of different etiologies enrolled in a dedicated prospective study. Thorough biochemical, clinical, hemodynamic, endoscopic, elastographic, and echo-Doppler work-up was performed in both studies. In study 1, no patients without cirrhosis developed HCC. Of 183 patients with cirrhosis, 14 of 28 (50.0%) with previous HCC recurred whereas 21 of 155 (13.5%) developed de novo HCC. Patients with recurrent and de novo HCCs had significantly higher liver fibrosis (LF) scores, portal pressure, and systemic inflammation than nonrecurrent HCC or patients never developing HCC. In recurrent/de novo HCC patients, tumor and nontumor ANGPT2 showed an inverse relationship with portal vein velocity (PVv; r = -0.412, P = 0.037 and r = -0.409, P = 0.047 respectively) and a positive relationship with liver stiffness (r = 0.526, P = 0.007; r = 0.525, P = 0.003 respectively). Baseline circulating VEGF and cirrhotic liver ANGPT2 were significantly related (r = 0.414, P = 0.044). VEGF increased during DAAs, remaining stably elevated at 3-month follow-up, when it significantly related with serum ANGPT2 (r = 0.531, P = 0.005). ANGPT2 expression in the primary tumor or in cirrhotic tissue before DAAs was independently related with risk of HCC recurrence (odds ratio [OR], 1.137; 95% confidence interval [CI], 1.044-1.137; P = 0.003) or occurrence (OR, 1.604; 95% CI, 1.080-2.382; P = 0.019). In study 2, DAA treatment (OR, 4.770; 95% CI, 1.395-16.316; P = 0.013) and large varices (OR, 3.857; 95% CI, 1.127-13.203; P = 0.032) were independent predictors of de novo HCC. Our study indicates that DAA-mediated increase of VEGF favors HCC recurrence/occurrence in susceptible patients, that is, those with more severe fibrosis and splanchnic collateralization, who already have abnormal activation in liver tissues of neo-angiogenetic pathways, as shown by increased ANGPT2. (Hepatology 2018; 00:000-000).


Apparent activity of 5-methylfurmethide at muscarinic receptors.
Apparent activity of 5-methylfurfuryltrimethylammonium iodide (5-methylfurmethide, 5-MFT) at muscarinic receptors of the ileal longitudinal muscle of the guinea-pig was investigated. These parameters were determined by the analysis of concentration-response data before and after fractional inactivation of receptors by phenoxybenzamine, an irreversible receptor blocker. The apparent ED50, dissociation constant (KA), and relative affinity (in comparison to acetylcholine, ACh) were 2.75 +/- 0.22 X 10(-8) M, 7.22 +/- 0.15 X 10(-7) M and 1.33 respectively. The relative intrinsic efficacy compared to ACh was 1.54.


Regulation of the development of mesencephalic dopaminergic systems by the selective expression of glial cell line-derived neurotrophic factor in their targets.
Glial cell line-derived neurotrophic factor (GDNF) has been shown to protect and restore dopamine (DA) neurons in injury models and is being evaluated for the treatment of Parkinson's disease. Nevertheless, little is known of its physiological role. We have shown that GDNF suppresses apoptosis in DA neurons of the substantia nigra (SN) postnatally both in vitro and during their first phase of natural cell death in vivo. Furthermore, intrastriatal injection of neutralizing antibodies augments cell death, suggesting that endogenous GDNF plays a role as a target-derived factor. Such a role would predict that overexpression of GDNF in striatum would increase the surviving number of SN DA neurons. To test this hypothesis, we used the tetracycline-dependent transcription activator (tTA)/tTA-responsive promoter system to create mice that overexpress GDNF selectively in the striatum, cortex, and hippocampus. These mice demonstrate an increased number of SN DA neurons after the first phase of natural cell death. However, this increase does not persist into adulthood. As adults, these mice also do not have increased dopaminergic innervation of the striatum. They do, however, demonstrate increased numbers of ventral tegmental area (VTA) neurons and increased innervation of the cortex. This morphologic phenotype is associated with an increased locomotor response to amphetamine. We conclude that striatal GDNF is necessary and sufficient to regulate the number of SN DA neurons surviving the first phase of natural cell death, but it is not sufficient to increase their final adult number. GDNF in VTA targets, however, is sufficient to regulate the adult number of DA neurons.


Prevalence of obesity, hypertension and diabetes among people living with HIV in South Africa: a systematic review and meta-analysis.
HIV has become a manageable chronic condition due to the success and scale-up of antiretroviral therapy (ART). Globally, South Africa has the highest number of people living with HIV (PLHIV) and research evidence indicates that countries with the highest burden of PLHIV have a substantial burden of obesity, hypertension (HPT) and type 2 diabetes (T2D). We sought to summarize the burden of these three common NCDs among PLHIV in South Africa. In this systematic review, multiple databases were searched for articles reporting on the prevalence of obesity, HPT, and T2D among PLHIV in South Africa published since journal inception until March 2022. A meta-analysis was conducted using random-effects models to obtain pooled prevalence estimates of the three NCDs. Heterogeneity was assessed using X2 test on Cochran's Q statistic. We included 32 studies, with 19, 22 and 18 studies reporting the prevalence of obesity, HPT, and T2D among PLHIV, respectively. The overall prevalence of obesity, HPT, and T2D was 23.2% [95% CI 17.6; 29.9], 25.5% [95% CI 15.6; 38.7], and 6.1% [95% CI 3.8; 9.7] respectively. The prevalence of obesity was significantly higher among women (P = 0.034) compared to men, however the prevalence of HPT and T2D did not differ by sex. The prevalence of each of the three NCDs did not differ significantly between rural, urban, and peri-urban areas. The prevalence of obesity and T2D was higher in studies conducted between 2013 and 2022 compared to studies conducted between 2000 and 2012, while the prevalence of HPT was higher between 2000 and 2012 compared to between 2013 and 2022. These findings suggest that South Africa is experiencing a syndemic of NCDs among people PLHIV highlighting the need to increase cost-effective interventions and management strategies that involve integrated HIV and NCD care in the South African setting.


Enhanced electroanalysis in lithium potassium eutectic (LKE) using microfabricated square microelectrodes.
Molten salts (MSs) are an attractive medium for chemical and electrochemical processing and as a result there is demand for MS-compatible analysis technologies. However, MSs containing redox species present a challenging environment in which to perform analytical measurements because of their corrosive nature, significant thermal convection and the high temperatures involved. This paper outlines the fabrication and characterization of microfabricated square microelectrodes (MSMs) designed for electrochemical analysis in MS systems. Their design enables precise control over electrode dimension, the minimization of stress because of differential thermal expansion through design for high temperature operation, and the minimization of corrosive attack through effective insulation. The exemplar MS system used for characterization was lithium chloride/potassium chloride eutectic (LKE), which has potential applications in pyrochemical nuclear fuel reprocessing, metal refining, molten salt batteries and electric power cells. The observed responses for a range of redox ions between 400 and 500 °C (673 and 773 K) were quantitative and typical of microelectrodes. MSMs also showed the reduced iR drop, steady-state diffusion-limited response, and reduced sensitivity to convection seen for microelectrodes under ambient conditions and expected for these electrodes in comparison to macroelectrodes. Diffusion coefficients were obtained in close agreement with literature values, more readily and at greater precision and accuracy than both macroelectrode and previous microelectrode measurements. The feasibility of extracting individual physical parameters from mixtures of redox species (as required in reprocessing) and of the prolonged measurement required for online monitoring was also demonstrated. Together, this demonstrates that MSMs provide enhanced electrode devices widely applicable to the characterization of redox species in a range of MS systems.


Characterization of Nigerian strains of Newcastle disease virus.
Newcastle disease virus was isolated from outbreaks of the disease in vaccinated and unvaccinated poultry flocks representing commercial and backyard farms in different parts of Nigeria. On characterization, all 12 isolates were found to be velogenic.


Alternation of anthelmintic treatments: a molecular evaluation for benzimidazole resistance in nematodes.
The evolution of benzimidazoles (BZ) resistance in Teladorsagia circumcincta was investigated in a controlled trial with lambs, submitted to different treatment regimens. Four paddocks were seeded with a T. circumcincta strain constituted by 25% of BZ-resistant nematodes. Ten permanent lambs were allocated to each paddock, from April to November in order to renew the contamination of pasture. Monthly, three tracer lambs were allocated in each paddock. BZ-resistant nematode frequency was determined (PCR diagnosis). The faecal egg count reduction test (permanent lambs) and the number of nematodes in lambs were also determined (permanent and tracer lambs). Four different regimens of treatments were performed: control, levamisole (a non-BZ drug), fenbendazole (a BZ drug), and an alternation of levamisole and fenbendazole every second treatment. The same protocol was repeated on two consecutive grazing seasons, increasing the number of treatments (3 in first year and 5 in second year). The proportions of BZ-resistant nematodes did not change during all the study in both the control and the levamisole paddocks, supporting an equal global fitness of BZ-resistant and susceptible nematodes. Thus, no reversion of BZ resistance is to be expected. In the alternated drug group and in the BZ treated group, BZ-resistant nematodes increased from 25% to 47% and to 78%, respectively. BZ resistance increased proportionally to the selective pressure (number of BZ treatments). The drug alternation is not a good solution to delay importantly the evolution of resistance when more than 25% of nematodes are BZ-resistant. This study is the first evaluation of BZ-resistance evolution (using individual genotyping) in controlled conditions. It showed that when a monogenic anthelmintic resistance is established at 25% in a sexually reproducing nematode population, it seems to be impossible to prevent its increase even when using limited number of BZ treatments.


The import of trust in regular providers to trust in cancer physicians among white, African American, and Hispanic breast cancer patients.
Interpersonal trust is an important component of the patient-doctor relationship. Little is known about patients' trust in the multiple providers seen when confronting serious illness. To characterize breast cancer patients' trust in their regular providers, diagnosing physicians, and cancer treatment team and examine whether high trust in one's regular provider confers high trust to cancer physicians. In-person interviews. 704 white, black, and Hispanic breast cancer patients, age 30 to 79, with a first primary in situ or invasive breast cancer who reported having a regular provider. We measure trust in: (1) regular provider, (2) diagnosing doctors, and (3) cancer treatment team. Other variables include demographic variables, preventive health care, comorbidities, time with regular provider, time since diagnosis, cancer stage, and treatment modality. Sixty-five percent of patients reported high trust in their regular provider, 84% indicated high trust in their diagnosing doctors, and 83% reported high trust in their treatment team. Women who reported high trust in their regular provider were significantly more likely to be very trusting of diagnosing doctors (OR: 3.44, 95% CI: 2.27-5.21) and cancer treatment team (OR: 3.09, 95% CI: 2.02-4.72 ). Black women were significantly less likely to be very trusting of their regular doctor (OR: 0.58, 95% CI: 0.38-0.88) and cancer treatment team (OR: 0.45, 95% CI: 0.25-0.80). English-speaking Hispanic women were significantly less trusting of their diagnosing doctors (OR: 0.29, 95% CI: 0.11-0.80). Our results suggest that patients are very trusting of their breast cancer providers. This is an important finding given that research with other populations has shown an association between trust and patient satisfaction and treatment adherence. Our findings also suggest that a trusting relationship with a regular provider facilitates trusting relationships with specialists. Additional work is needed to increase interpersonal trust among black women.


Chronic pancreatitis in a patient with malnutrition due to anorexia nervosa.
Both acute and chronic pancreatitis are associated with eating disorders, including malnutrition found in anorexia, bulimia, and major depression. We report a case of a female patient suffering from severe malnutrition and anorexia with repeated attacks of pancreatic pain and an enlarging cystic lesion in the pancreatic head. Due to a progressively enlarging lesion on CT, a pancreaticoduodenectomy was performed. Histology demonstrated chronic pancreatitis. The pathogenesis of chronic pancreatitis remains to be well defined. There is evidence that an imbalance between oxidative stress and antioxidant capacity results in pancreatic inflammation and activation of periacinar myofibroblasts. It has been demonstrated that protein energy malnutrition is associated with increased levels of proinflammatory cytokines as well as pancreatic acinar cell damage and ductal disruption. Furthermore, it has been shown that protein energy malnutrition including anorexia nervosa is associated with a depleted antioxidant status. Thus there is a possible pathogenic basis for severe malnutrition leading to chronic pancreatitis. Our patient underwent surgery based on the presumption that she had a symptomatic cystic neoplasm. Chronic pancreatitis was demonstrated. Patients presenting with malnutrition and recurrent epigastric pain should be investigated for pancreatic pathology and the possibility of pancreatitis and the presence of pseudocysts entertained.


Hemodialyzer reuse: practices in the United States and implication for infection control.
Reuse of hemodialyzers has continued to increase dramatically. In 1983, 52% of the hemodialysis centers were reusing hemodialyzers. Use of hollow fiber hemodialyzers has paralleled this increase in hemodialyzer reuse with 63% of the hemodialysis centers reporting exclusive use of this type of hemodialyzer in 1983. Although reuse of hemodialyzers has not been associated with increased incidence of hepatitis B infection or pyrogenic reactions, an outbreak of nontuberculous mycobacteria in 27 chronic hemodialysis patients was probably associated with water used to reprocess dialyzers. Nontuberculous mycobacteria were isolated from the water in 83% of 115 hemodialysis centers surveyed across the United States and could constitute a potential infection risk because of the organisms' greater germicide resistance than most other naturally occurring water bacteria. Two percent formaldehyde is not an effective germicide for high level disinfection of hemodialyzers. Reprocessed hemodialyzers should be disinfected with 4% formaldehyde or an equivalent disinfectant.


DBIRD complex integrates alternative mRNA splicing with RNA polymerase II transcript elongation.
Alternative messenger RNA splicing is the main reason that vast mammalian proteomic complexity can be achieved with a limited number of genes. Splicing is physically and functionally coupled to transcription, and is greatly affected by the rate of transcript elongation. As the nascent pre-mRNA emerges from transcribing RNA polymerase II (RNAPII), it is assembled into a messenger ribonucleoprotein (mRNP) particle; this is the functional form of the nascent pre-mRNA and determines the fate of the mature transcript. However, factors that connect the transcribing polymerase with the mRNP particle and help to integrate transcript elongation with mRNA splicing remain unclear. Here we characterize the human interactome of chromatin-associated mRNP particles. This led us to identify deleted in breast cancer 1 (DBC1) and ZNF326 (which we call ZNF-protein interacting with nuclear mRNPs and DBC1 (ZIRD)) as subunits of a novel protein complex--named DBIRD--that binds directly to RNAPII. DBIRD regulates alternative splicing of a large set of exons embedded in (A + T)-rich DNA, and is present at the affected exons. RNA-interference-mediated DBIRD depletion results in region-specific decreases in transcript elongation, particularly across areas encompassing affected exons. Together, these data indicate that the DBIRD complex acts at the interface between mRNP particles and RNAPII, integrating transcript elongation with the regulation of alternative splicing.


Oxidation-reduction properties of rat liver cytochromes P-450 and NADPH-cytochrome p-450 reductase related to catalysis in reconstituted systems.
A series of equilibrium and kinetic measurements involving the oxidation-reduction properties of purified rat liver NADPH-cytochrome P-450 reductase and eight different purified rat liver cytochromes P-450 (P-450s) were carried out. Apparent spin states of P-450 iron were determined in the absence and presence of a number of known substrates by using second-derivative and conventional near-UV absorbance spectroscopy. Many of the substrates examined did not produce significant changes in the apparent iron spin state, even when binding could be demonstrated with equilibrium dialysis. Further, the spin state was not correlated to catalytic activity of the P-450s in reconstituted enzyme systems. The oxidation-reduction potentials were determined for the ferric/ferrous couples of each of the eight P-450s in the presence and absence of known substrates, as well as other proteins suspected of altering the potentials. The midpoint potential (Em,7) ranged from -350 to -289 mV for the P-450s under these conditions. In some cases Em,7 was raised with the addition of substrates, but the extent of the increase was no greater than +33 mV. The Em,7 of one P-450 (P-450 beta NF/ISF-G) was not changed significantly when the fraction of high-spin iron varied between 11 and 67%. Steady-state spectral studies provided evidence for the accumulation of an oxygenated ferrous intermediate (or a derived product) of one P-450 (P-450PB-B) in the presence of a substrate, cyclohexane. Studies on the donation of electrons from cytochrome b5 and a series of dyes to this complex suggest that it has an effective Em,7 (for reduction) of approximately +50 mV. In studies with one of the P-450s, steady-state spectral studies indicated that the three-electron-reduced form of NADPH-P-450 reductase accumulates, consistent with the view that this form of the reductase is involved in the reduction of P-450 from the ferric to the ferrous state.


Error Characterization and Statistical Modeling Improves Circulating Tumor DNA Detection by Droplet Digital PCR.
Droplet digital PCR (ddPCR) is a widely used and sensitive application for circulating tumor DNA (ctDNA) detection. As ctDNA is often found in low abundance, methods to separate low-signal readouts from noise are necessary. We aimed to characterize the ddPCR-generated noise and, informed by this, create a sensitive and specific ctDNA caller. We built 2 novel complimentary ctDNA calling methods: dynamic limit of blank and concentration and assay-specific tumor load estimator (CASTLE). Both methods are informed by empirically established assay-specific noise profiles. Here, we characterized noise for 70 mutation-detecting ddPCR assays by applying each assay to 95 nonmutated samples. Using these profiles, the performance of the 2 new methods was assessed in a total of 9447 negative/positive reference samples and in 1311 real-life plasma samples from colorectal cancer patients. Lastly, performances were compared to 7 literature-established calling methods. For many assays, noise increased proportionally with the DNA input amount. Assays targeting transition base changes were more error-prone than transversion-targeting assays. Both our calling methods successfully accounted for the additional noise in transition assays and showed consistently high performance regardless of DNA input amount. Calling methods that were not noise-informed performed less well than noise-informed methods. CASTLE was the only calling method providing a statistical estimate of the noise-corrected mutation level and call certainty. Accurate error modeling is necessary for sensitive and specific ctDNA detection by ddPCR. Accounting for DNA input amounts ensures specific detection regardless of the sample-specific DNA concentration. Our results demonstrate CASTLE as a powerful tool for ctDNA calling using ddPCR.


Nutlin-3 up-regulates the expression of Notch1 in both myeloid and lymphoid leukemic cells, as part of a negative feedback antiapoptotic mechanism.
The small molecule inhibitor of the MDM2/p53 interaction Nutlin-3 significantly up-regulated the steady-state mRNA and protein levels of Notch1 in TP53(wild-type) (OCI, SKW6.4) but not in TP53(deleted) (HL-60) or TP53(mutated) (BJAB) leukemic cell lines. A direct demonstration that NOTCH1 was a transcriptional target of p53 in leukemic cells was obtained in experiments carried out with siRNA for p53. Moreover, inhibition of Notch1 expression using Notch1-specific siRNA significantly increased cytotoxicity in TP53(wild-type) leukemic cells. Of note, Nutlin-3 up-regulated Notch1 expression also in primary TP53(wild-type) B-chronic lymphocytic leukemia (B-CLL) cells and the combined use of Nutlin-3 plus pharmacological gamma-secretase inhibitors of the Notch signaling showed a synergistic cytotoxicity in both TP53(wild-type) leukemic cell lines and primary B-CLL cells. A potential drawback of gamma-secretase inhibitors was their ability to enhance osteoclastic maturation of normal circulating preosteoclasts induced by RANKL + M-CSF. Notwithstanding, Nutlin-3 completely suppressed osteoclastogenesis irrespective of the presence of gamma-secretase inhibitors. Taken together, these data indicate that the p53-dependent up-regulation of Notch1 in response to Nutlin-3 represents an antiapoptotic feedback mechanism able to restrain the potential therapeutic efficacy of Nutlin-3 in hematologic malignancies. Therefore, therapeutic combinations of Nutlin-3 + gamma-secretase inhibitors might potentiate the cytotoxicity of Nutlin-3 in p53(wild-type) leukemic cells.


Surgery in the very elderly.
The very elderly who underwent major surgical procedures had longer hospital stays and higher total charges than the younger age group in the majority of DRGs studied. Higher mortality rates in the very elderly for most of the DRGs were found to be associated with emergency surgery for conditions that carry very high mortality rates when surgery is not performed. Surgery in the very elderly, then, although from 30% to 40% more costly in terms of hospital days and charges, should not be withheld on the basis of age alone. Careful assessment of risks to each individual patient should be continued and can result in acceptable mortality rates.


Detection of resistance to isoniazid by denaturing gradient-gel electrophoresis DNA sequencing in Mycobacterium tuberculosis clinical isolates.
Isoniazid (INH) resistance was genotypically assessed in 104 (37 INH-susceptible, 67 INH-resistant) genetically unrelated Mycobacterium tuberculosis strains cultured in North Italy. The PCR products of selected regions of the katG gene, the oxyR-ahpC intergenic region, and the inhA regulatory region were analyzed utilizing the double gradient-denaturing gradient gel electrophoresis (DG-DGGE) technique and confirmed by DNA sequencing. Mutations were detected in 61 (91%) of the INH-resistant strains, the relative frequency of the mutations being 65.7% in katG, 23.9% in oxyR-ahpC, and 13.4% in inhA. Previously described alterations, invariably associated with drug resistance, accounted for 95.1% of the mutations. No alterations were found in the INH-susceptible strains. DG-DGGE analysis and DNA sequencing were equally sensitive, but the former is cheaper, easier and more robust. Rapid genotypic assessment of INH resistance by means of the methodology described here could reasonably be used in clinical mycobacteriology laboratories.


Healthcare Human Resources: Trends and Demand in Saudi Arabia.
This paper estimates the impact of policies on the current status of Healthcare Human Resources (HHR) in Saudi Arabia and explores the initiatives that will be adopted to achieve Saudi Vision 2030. Retrospective time-series data from the Ministry of Health (MOH) and statistical yearbooks between 2003 and 2015 are analyzed to identify the impact of these policies on the health sector and the number of Saudi and non-Saudi physicians, nurses and allied health specialists employed by MOH, Other Government Hospitals (OGH) and Private Sector Hospitals (PSH). Moreover, multiple regressions are performed with respect to project data until 2030 and meaningful inferences are drawn. As a local supply of professional medical falls short of demand, either policy to foster an increase in supply are adopted or the Saudization policies must be relaxed. The discrepancies are identified in terms of a high rate of non-compliance of Saudization in the private sector and this is being countered with alternative measures which are discussed in this paper. The study also analyzed the drivers of HHR demand, supply and discussed the research implications on policy and society. The findings suggest that the 2011 national Saudization policy yielded the desired results mostly regarding allied health specialists and nurses. This study will enable decision-makers in the healthcare sector to measure the effectiveness of the new policies and, hence, whether to continue in implementing them or to revise them.


Rapid start-up of an innovative pilot-scale staged PN/A continuous process for enhanced nitrogen removal from mature landfill leachate via robust NOB elimination and efficient biomass retention.
The start-up and stable operation of partial nitritation-anammox (PN/A) treatment of mature landfill leachate (MLL) still face challenges. This study developed an innovative staged pilot-scale PN/A system to enhance nitrogen removal from MLL. The staged process included a PN unit, an anammox upflow enhanced internal circulation biofilm (UEICB) reactor, and a post-biofilm unit. Rapid start-up of the continuous flow PN process (full-concentration MLL) was achieved within 35 days by controlling dissolved oxygen and leveraging free ammonia and free nitrous acid to selectively suppress nitrite-oxidizing bacteria (NOB). The UEICB was equipped with an annular flow agitator combined with the enhanced internal circulation device of the guide tube, which achieved an efficient enrichment of Candidatus Kuenenia in the biofilm (relative abundance of 33.4 %). The nitrogen removal alliance formed by the salt-tolerant anammox bacterium (Candidatus Kuenenia) and denitrifying bacteria (unclassified SBR1031 and Denitratisoma) achieved efficient nitrogen removal of UEICB (total nitrogen removal percentage: 90.8 %) and at the same time effective treatment of the refractory organic matter (ROM). The dual membrane process of UEICB fixed biofilm combined with post-biofilm is effective in sludge retention, and can stably control the effluent suspended solids (SS) at a level of less than 5 mg/L. The post-biofilm unit ensured that effluent total nitrogen (TN) remained below the 40 mg/L discharge standard (98.5 % removal efficiency). Compared with conventional nitrification-denitrification systems, the staged PN/A process substantially reduced oxygen consumption, sludge production, CO2 emissions and carbon consumption by 22.8 %, 67.1 %, 87.1 % and 87.1 %, respectively. The 195-day stable operation marks the effective implementation of the innovative pilot-scale PN/A process in treating actual MLL. This study provides insights into strategies for rapid start-up, robust NOB suppression, and anammox biomass retention to advance the application of PN/A in high-ammonia low-carbon wastewater.


Bilateral paramedian thalamic syndrome after infection.
Although bilateral paramedian thalamic infarctions occur more frequently in adults than in children, they are rare entities at any age. The syndrome is thought to result from occlusion of the artery of Percheron, which arises as a common trunk from one of the posterior cerebral arteries to supply both paramedian thalamic regions. We describe two children with acute ischemic infarction involving both paramedian thalami developing after infection. The first patient developed mutism with ataxia after chicken pox infection. The second child developed headache, somnolence, agitation, and speech dysfunction following an upper respiratory tract infection. Bilateral thalamic lesions were documented on magnetic resonance imaging of both children. Bilateral infarctions of the paramedian thalamus may result in severe illness and impairment. Common clinical manifestations include disorientation, confusion, hypersomnolence, deep coma and "coma vigil," or akinetic mutism (awake unresponsiveness), as well as severe memory impairment.


Mutation in POLR3K causes hypomyelinating leukodystrophy and abnormal ribosomal RNA regulation.
To identify the genetic cause of hypomyelinating leukodystrophy in 2 consanguineous families. Homozygosity mapping combined with whole-exome sequencing of consanguineous families was performed. Mutation consequences were determined by studying the structural change of the protein and by the RNA analysis of patients' fibroblasts. We identified a biallelic mutation in a gene coding for a Pol III-specific subunit, POLR3K (c.121C>T/p.Arg41Trp), that cosegregates with the disease in 2 unrelated patients. Patients expressed neurologic and extraneurologic signs found in POLR3A- and POLR3B-related leukodystrophies with a peculiar severe digestive dysfunction. The mutation impaired the POLR3K-POLR3B interactions resulting in zebrafish in abnormal gut development. Functional studies in the 2 patients' fibroblasts revealed a severe decrease (60%-80%) in the expression of 5S and 7S ribosomal RNAs in comparison with control. These analyses underlined the key role of ribosomal RNA regulation in the development and maintenance of the white matter and the cerebellum as already reported for diseases related to genes involved in transfer RNA or translation initiation factors.