Tuning and matching networks for MR imaging and spectroscopy.
The authors briefly discuss four tuning and matching networks and their application range in magnetic resonance (MR) imaging and MR spectroscopy. Analytic expressions for the network elements needed to tune and match an MR imaging or spectroscopy probe for a given coil impedance are given, and the connection to the "standard" two-element capacitive matching networks is shown. The power loss factor and the phase shift between the input current and the load current are explicitly calculated for each of the four networks. The merits of the different matching networks are then discussed in the context of a general-purpose license plate coil commonly used in MR imaging.


Genetics of acute and chronic pancreatitis.
Acute pancreatitis, recurrent acute pancreatitis (RAP) and chronic pancreatitis are interrelated and progressive inflammatory disorders of the pancreas with highly variable and complex susceptibility, severity and outcomes. The role of genetics in acute pancreatitis, RAP and progression to chronic pancreatitis within a new framework is needed. The first genome-wide association study in the pancreas has been published with genome-wide significance linked with noncoding variants at the PRSS1-PRSS2 locus on chromosome seven and the CLDN2 locus on the X chromosome. A new personalized medicine paradigm is being considered to facilitate organization of genetic and other susceptibility risk compared with the risk of disease progression or resolution and risk of complications. A new framework for organizing multiple, complex data sets is emerging. The role of genetics in the context of other variables is important in understanding susceptibility to RAP and in the modification of disease severity and progression to chronic pancreatitis. Questions of when to order testing, what to order and how to use the data in real time remains an area for future research and development.


Decreased interleukin 1 activity in culture supernatant of lipopolysaccharide stimulated monocytes from patients with liver cirrhosis and hepatocellular carcinoma.
Immunoregulatory function of peripheral blood monocytes was studied in patients with hepatocellular carcinoma (HCC) and liver cirrhosis (LC), by assaying interleukin 1 (IL-1) and prostaglandin E2 (PGE2) in the culture supernatant of lipopolysaccharide-stimulated monocytes. IL-1 activity of the monocyte culture supernatant without indomethacin was decreased in patients with HCC and LC, compared with that of controls. The activity was lower in patients with HCC than that in those with LC. The PGE2 content of the culture supernatant of monocytes from patients with LC and HCC was increased, compared to normal controls. To avoid the effect of PGE2 on the IL-1 assay, we cultured the monocytes with addition of indomethacin and assayed IL-1 activity in the culture supernatant. As a result, monocyte IL-1 production was increased in patients with HCC and LC, compared with normal controls. The decrease in IL-1 activity of the supernatant without indomethacin of patients with LC and HCC was considered to be due to increased secretion of PGE2 by the monocytes. Therefore, monocytes from patients with HCC and LC had an increased capacity of secreting both IL-1 and PGE2 over normal controls, but the effect of the suppressor function (PGE2 secretion) dominated in these patients.


Device Safety Implications of the Clinical Data Leading to US Food and Drug Administration Approval of Soft-Tissue Fillers: A Systematic Review.
The US Food and Drug Administration (FDA) recently issued a safety warning regarding soft-tissue fillers (STFs) based on the risk of blindness and facial necrosis. To examine the quality of evidence leading to FDA approval of STFs. All original approvals for STFs were mined using the publicly available FDA database. The Cochrane Collaboration's risk of bias assessment tool was applied to all randomized clinical trials (RCTs). A total of 14 STF approvals were identified. Of those, 10 pivotal studies (71%) were RCTs and 9 (60%) were masked. The median number of patients per trial was 144 (range, 30-439). Eleven of 12 studies (92%) met their primary end point. Ten of 14 trials (71%) involved injections solely of the nasolabial folds or cheeks; only 4 trials involved treatment of other facial regions. All 10 RCTs had an unclear risk of selection bias. Only 2 RCTs reported exclusions and attrition. Safety warnings relate more to the off-label use of STFs, which has not been sufficiently studied prospectively. Although STFs remain a safe device, with approval based mostly on RCT outcomes, implementation of unique device identifiers and greater use of physician-led registries would ensure physician, consumer, and regulatory confidence in STF safety.


[EPIDURAL ANALGESIA FOR INTRAOPERATIVE AND POSTOPERATIVE CARE IN NEWBORNS].
Anesthesia care in newborns has to be complex, balanced and safe. Nowadays epidural analgesia (EA) in neonates during intra- and postoperative period is widely used in Russia. Modern EA techniques imply the installation of a catheter into epidural space at lumbar or thoracic level as well as different approach to local anesthetics dosage. Newborns have special anatomy, physiology and pharmacodynamics which have to be taken in mind when EA is used. At the present moment Ropivacine (2 mg/ml) is approved for peripheral nerve blocks in newborns.


Determination of Appropriate Umbilicus Position during Abdominoplasty in Male Patients.
Abdominoplasty techniques are well documented. The ideal position of the umbilicus has, however, received limited attention. Unfortunately, umbilicus position is not universally agreed upon in male cosmetic abdominoplasty. This study was conducted to determine the ideal umbilicus anthropometric measurements in young men, and the relationships between umbilical position and anterior trunk and torso reference points that may be applicable to intraoperative positioning. It is aimed also at investigating whether umbilicus position would be more accurately determined by considering nipple position instead of the abdominal crease, as recently proposed. Several anthropometric measurements of various anterior abdominal and thoracic landmarks were conducted on 60 young and middle-aged male volunteers and 30 cadavers at São Paulo city. All statistical analysis was completed using Stata software. Of all the measured reference points, a much stronger correlation (0.513) was demonstrated between umbilicus-anterior axillary fold (U-AX) and inter-nipple (N-N) distances with a constant golden number ratio relationship (N-N = U-AX × 0.618) compared with the weak correlation of 0.034 between umbilicus-xiphisternum and umbilicus-abdominal crease. In 75% of volunteers, the calculated U-AX was within ±3 cm of actual measurement, and in 33.33% within ±1 cm. U-AX = 1.618 × N-N equation is more predictive of adequate umbilicus repositioning during abdominoplasty in male patients. Chest and abdomen of men are a single aesthetic unit. Proper positioning of the nipples and umbilicus, as well as harmonious abdominal and torso proportions are critical for an optimal final aesthetic outcome.


[Image diagnosis of vasculitis--ultrasound].
Ultrasound is widely used for the diagnosis and follow-up of patients with many kinds of vasculitis. Transcutaneous or transesophageal two-dimensional ultrasound images can depict vascular wall lesions associated with vasculitis such as stenosis, aneurysm or wall thickening. Pulsed Doppler and color Doppler are useful for the assessment of stenoses and changes in blood flow. Recent progress in intravascular ultrasound apparatus made it possible to more precisely assess morphological changes in vascular wall. Intravascular ultrasound of a 27-year old male patient suffering from inferior myocardial infarction with right coronary aneurysms revealed calcification and fibrosis of the intima and early development of atherosclerosis in both right and left coronary arteries suggestive of the sequelae of Kawasaki disease.


A Method for Islet Transplantation to the Omentum in Mouse.
Islet transplantation has been proposed to be a potential treatment for type 1 diabetes. Recent compelling evidence indicates that intravascular islet infusion is far from ideal and therefore, the omentum is re-emerging as a potentially valuable site for islet transplantation. This experiment requires the isolation of high quality islets and the implantation of the islets to the diabetic recipients. Transplantation to the omentum requires surgical steps that can be better demonstrated visually. Here, the detailed steps for this procedure are presented. Two methods of mixing the isolated islets with hydrogel before placing the mixture into the omental pouch of diabetic mice are described here. Different hydrogels are used for the different conditions. Blood glucose levels of diabetic mouse recipients of syngeneic islets in the omentum were monitored for up to 35 days. Some animals were sacrificed after 14 days to perform immuno-histochemical analysis. This pre-clinical transplantation approach can be used as preliminary data leading up to translation to clinical transplantation.


Isolation and identification of mycoplasmas from the nasal cavity of sheep.
Mycoplasmas isolated from the nasal cavity of sheep in a ram test station were examined to determine their identity and prevalence. Specimens were obtained for mycoplasmal culture in 1980, 1982, and 1983 from 558 sheep, and mycoplasmas were isolated from 630 specimens from 320 sheep (57.3%). The isolates were characterized and differentiated into groups on the basis of sensitivity to digitonin, fermentation of glucose, and hydrolysis of arginine. Isolates in some groups were further characterized by use of additional diagnostic media, and their identity was confirmed by agglutination or growth inhibition with antiserum prepared from reference mycoplasmas. Of the 320 sheep with mycoplasmas, 293 had Mycoplasma ovipneumoniae, 12 had M arginini, and 1 had M capricolum. Two sheep had Acholeplasma spp, and 3 sheep had unidentified Mycoplasma spp. The remaining 9 sheep had M ovipneumoniae in combination with Acholeplasma spp (n = 3), M arginini (n = 3), M capricolum (n = 2), and an unidentified Mycoplasma spp (n = 1). The biochemical reactions of the M ovipneumoniae from the 293 sheep were similar, but varied in the degree of growth and fermentation in the basal medium containing glucose. The high prevalence of M ovipneumoniae indicated that it may be commensal in the upper respiratory tract of healthy sheep.


Clinicopathologic Features and Treatment Characteristics of Congenital Corneal Opacity Infants and Children Aged 3 Years or Less: A Retrospective Single Institution Analysis.
In this retrospective single institution study, we investigated the clinicopathologic features and treatment characteristics of 90 patients with congenital corneal opacities (CCO) (117 eyes) who were 3 years and younger and treated at our hospital. We reviewed the clinical data of patients with CCO who presented for the first time for treatment at our hospital between January 1, 2017, and December 31, 2017. CCO were classified using the "STUMPED" (Sclerocornea, Tears in Descement's membrane, Metabolic, Peters, Endothelial dystrophy and Dermoid) method and confirmed by pathological examination. -Results: Seventy percent of the patients had unilateral CCO. Iridocorneal adhesions (61 eyes, 52.1%) and cataracts (22 eyes, 18.8%) were the 2 most common ocular abnormalities. Systemic abnormalities were present in 5 patients (5.6%), including growth retardation (4 patients) and congenital brain defects (1 patient). Eighty-five eyes (72.6%) underwent penetrating keratoplasty (PK), and lamellar keratoplasty (LK) was performed in 30 (25.6%) eyes. Forty-seven (95.9%) eyes with Peters anomaly and all 16 eyes with sclerocornea received PK, and all 24 eyes with dermoids were treated with LK. Our study demonstrates that CCO has varied manifestations in infants and young children in China. A thorough medical history, careful clinical examination, and the use of accessory examinations such as ultrasound biomicroscopy are critical for the accurate diagnosis and classification of CCO and to provide guidance on therapeutic choices.


Comparing rapid assessments of delay discounting with real and hypothetical rewards in children.
Recent advances in assessment methodology have resulted in a highly efficient procedure for obtaining delay discounting rates for adults: a 5-trial adjusting delay task (ADT-5) examining intertemporal choice for hypothetical rewards. The low participant burden of this task makes it potentially useful for children, with whom delay discounting research is relatively limited. However, it is unknown whether results from this task match choice for real rewards. The present study assessed delay discounting for real and hypothetical monetary rewards using a modified ADT-5 with 9 children admitted to a psychiatric day treatment program. Participants completed up to 3 tasks with each reward type in alternating order. No difference in discounting rate, via log(k), was observed between the first task of each reward type. This finding was replicated across subsequent tasks for the subset of participants (n = 6) who completed all 6 tasks. However, delay discounting of real and hypothetical rewards was not found to be statistically equivalent. These results suggest that a modified ADT-5 using hypothetical rewards may be a viable option for assessing delay discounting in children with psychiatric diagnoses, but additional research is needed to explicitly examine whether hypothetical and real rewards are discounted equivalently in this population.


Adiponectin overexpression in C2C12 myocytes increases lipid oxidation and myofiber transition.
Metabolic syndrome and obesity have detrimental effects on the metabolic function of the skeletal muscle. Mounting evidence indicates that patients with those conditions may present an increased ratio of glycolytic to oxidative fibers associated with a decrease in oxidative capacity. In this regard, adiponectin, a hormone mainly secreted by adipocytes that regulates glucose and lipid metabolism, has emerged as a myokine that could play an important role in this process. We aimed to investigate whether adiponectin overexpression in skeletal muscle might be a local protective mechanism, favoring fatty acid utilization. To that end, we generated an in vitro model of myocytes with upregulated endogenous adiponectin using a lentiviral carrier. We demonstrated that the adiponectin-transduced myocytes were able to produce and secrete fully functional adiponectin complexes. Adiponectin overexpression remarkably upregulated the mRNA level of myogenic regulatory factors as well as genes implicated in lipolysis (HSL, ATGL) and cellular and mitochondrial fatty acid transport (LPL, CD36, CPT1B). This was accompanied by increased isoproterenol-induced lipolysis and β-oxidation and reduced lipogenesis, whereas insulin-stimulated glucose uptake was unaltered in transduced myocytes. Lastly, the relative expression of the more glycolytic myofibers (MyHC IIb) compared to the more oxidative ones (MyHC I) was notably reduced. Our results showed that the released adiponectin acted in an autocrine/paracrine manner, increasing lipid oxidation in myocytes and leading to a transition of myofibers from the glycolytic to the oxidative type. In conclusion, muscle adiponectin overexpression might be a way to relieve muscle diseases caused by oxidative muscle fiber deficiency.


Rats (Rattus norvegicus and Rattus losea) harboring Seoul hantavirus in Qingyuan, southern China: a survey during 2011-2013.
Hemorrhagic fever with renal syndrome (HFRS) is a zoonotic disease, which threatens public health and its incidence has increased sharply up to the present time in southern China. A survey of HFRS including in both the natural hosts and humans conducted in Qingyuan, southern China, during 2011-2013 revealed that one, two and seven confirmed cases of HFRS occurred in 2011, 2012 and 2013, respectively. Rodent densities ranged 1.73%-12.2% and Seoul hantavirus was detected by RT-PCR only in Rattus norvegicus and Rattus losea. The positive rate in humans was 0.95% serologically and 0.94% with IIFA in 2011 and 2013, respectively. DNA fragments detected in Rattus norvegicus and Rattus losea were highly homologous with those of Seoul hantavirus HB55 (96.2%) and L99 (95.1%), respectively. Thus HFRS is becoming an emerging and dangerous disease in southern China and it is necessary to further perform molecular characterization of strains isolated from rodents and humans.


Discovery of Novel Oleamide Analogues as Brain-Penetrant Positive Allosteric Serotonin 5-HT2C Receptor and Dual 5-HT2C/5-HT2A Receptor Modulators.
The serotonin 5-HT2A receptor (5-HT2AR) and 5-HT2CR localize to the brain and share overlapping signal transduction facets that contribute to their roles in cognition, mood, learning, and memory. Achieving selective targeting of these receptors is challenged by the similarity in their 5-HT orthosteric binding pockets. A fragment-based discovery approach was employed to design and synthesize novel oleamide analogues as selective 5-HT2CR or dual 5-HT2CR/5-HT2AR positive allosteric modulators (PAMs). Compound 13 (JPC0323) exhibited on-target properties, acceptable plasma exposure and brain penetration, as well as negligible displacement to orthosteric sites of ∼50 GPCRs and transporters. Furthermore, compound 13 suppressed novelty-induced locomotor activity in a 5-HT2CR-dependent manner, suggesting 5-HT2CR PAM, but not 5-HT2AR, activity at the level of the whole organism at the employed doses of 13. We discovered new selective 5-HT2CR PAMs and first-in-class 5-HT2CR/5-HT2AR dual PAMs that broaden the pharmacological toolbox to explore the biology of these vital receptors.


Adolescents with severe chronic fatigue syndrome can make a full recovery.
The needs of children and adolescents severely affected by chronic fatigue syndrome, myalgic encephalomyelitis (CFS/ME) are currently inadequately addressed in the UK. Sadly, there are few specialists addressing the needs of these patients who are primarily bed-bound, wheelchair users or who can only leave home on an infrequent basis. Uncertainty about what to offer as well of a lack of funding may play a part. Action for Young people with ME (AYME) suggests that at least 350 severely affected children/adolescents are receiving little or inadequate care to help them overcome this debilitating illness. This case report illustrates how recovery can occur with pragmatic rehabilitation combined with a committed compassionate family based approach.


Molecular analysis of the hydrogenosomal ferredoxin of the anaerobic protist Trichomonas vaginalis.
We have determined the primary structure of the [2Fe-2S]ferredoxin of the anaerobic protist Trichomonas vaginalis. This protein, situated in the hydrogenosome, is composed of 93 amino acids. A comparison of T. vaginalis ferredoxin with greater than 80 other ferredoxins shows the closest similarity to [2Fe-2S]putidaredoxin of the aerobic bacterium Pseudomonas putida and a lesser one to mitochondrial [2Fe-2S]ferredoxins of vertebrates. This similarity is reflected in the overall primary structure and in the spacing of cysteine residues coordinating the iron-sulfur center. The primary structure, but not the environment of the iron-sulfur center, also shows similarity with [2Fe-2S]ferredoxins of photosynthetic organisms and halobacteria. We have cloned and analyzed the T. vaginalis ferredoxin gene. The gene is present in a single copy and devoid of introns. It gives rise to a transcript with unusually short 5' and 3' untranslated regions of 16 and 18 nucleotides, respectively. DNA sequence analysis of the gene predicts an additional 8 amino acids at the amino terminus which are absent from the purified protein. This amino-terminal region of the protein is characterized by properties typical of mitochondrial presequences.


Globular chitosan prolongs the effective duration time and decreases the acute toxicity of botulinum neurotoxin after intramuscular injection in rats.
Botulinum neurotoxin (BoNT) is used for an increasing number of neurological and non-neurological indications and disorders. Since the duration of action of this neurotoxin is limited, the goal of the work was to improve the pharmacological time course of BoNT. We explored the effect of several polysaccharides on the duration of action of BoNT/A1 in rat electromyography. The formulation of BoNT/A1 containing globular chitosan increased the threshold stimulation intensity almost 2 times in 30 days after injection if compared with the baseline threshold. However, conventional linear chitosan, heparin and hyaluronic acid did not have such an effect. In addition, we compared the effectiveness of different doses of BoNT/A1 (25, 50, 75, and 100 U) with globular chitosan and compared the acute toxicity of this formulation with that of BoNT/A1 in physiological saline after intramuscular injection. The results demonstrated that the dose 25 U of BoNT/A1 with globular chitosan was both effective and safe for animals after intramuscular injection. The assessed median lethal dose (LD50) for intramuscular injection in rats was 1.4 times higher for a combination of BoNT/A1 with globular chitosan than that for a solution of BoNT/A1 in physiological saline. Thus, the results of our study have provided evidence that intramuscular injection of the formulation of BoNT/A1 (25 U) containing globular chitosan in rats is safe and significantly prolongs the effective duration time of BoNT/A1.


Thirty-Two-Year Success of Dental Implants in Periodontally Compromised Dentition.
Osseointegrated dental implants have become an integral factor in the replacement of missing teeth. These implants have demonstrated long-term success for periodontally compromised patients, who require a high level of success to maintain a sense of optimism. This case report offers 32-year results of implant treatment in the maxilla and 25-year results in the mandible. Some maxillary implants suffered a few threads of bone loss, but all seven mandibular implants met Albrektsson's definition of success after 25 years. The purpose of this case report is to provide evidence that further supports the findings of osseointegrated implant treatment's longevity and success, as reported in other studies.


Stereochemical elucidation of streptorubin B.
Streptorubin B is a structurally remarkable member of the prodiginine group of antibiotics produced by several actinobacteria, including the model organism Streptomyces coelicolor A3(2). Transannular strain within the pyrrolophane structure of this molecule causes restricted rotation that gives rise to the possibility of (diastereomeric) atropisomers. Neither the relative nor the absolute stereochemistry of streptorubin B is known. NOESY NMR experiments were used to define the relative stereochemistry of the major atropisomer of streptorubin B·HCl in solution as anti. We exploited this finding together with our knowledge of streptorubin B biosynthesis in S. coelicolor to determine the absolute stereochemistry of the anti atropisomer. 2-Undecylpyrrole stereoselectively labeled with deuterium at C-4' was synthesized and fed to a mutant of S. coelicolor, which was unable to produce streptorubin B because it was blocked in 2-undecylpyrrole biosynthesis, and in which the genes responsible for the last two steps of streptorubin B biosynthesis were overexpressed. (1)H and (2)H NMR analysis of the stereoselectively deuterium-labeled streptorubin B·HCl produced by this mutasynthesis strategy allowed us to assign the absolute stereochemistry of the major (anti) atropisomer as 7'S. HPLC analyses of streptorubin B isolated from S. coelicolor on a homochiral stationary phase and comparisons with streptorubin B derived from an enantioselective synthesis showed that the natural product consists of an approximately 88:7:5 mixture of the (7'S, anti), (7'S, syn), and (7'R, anti) stereoisomers.


Waveguide-based single-pixel up-conversion infrared spectrometer.
A periodically poled lithium niobate (PPLN) waveguide-based single-pixel up-conversion infrared spectrometer was demonstrated. Sum-frequency generation between a 1.5-microm-band scanning pump laser and a 1.3-microm-band signal generated visible radiation which was detected by a silicon single-photon detector. The up-conversion spectrometer's sensitivity was two-orders-of-magnitude higher than that of a commercial optical spectrum analyzer.


Undetected gynaecological disorders in women with renal disease.
Women with chronic renal disease (CRD) who are on dialysis or have a functioning renal transplant are typically stoical in their attitude towards other health problems. We undertook a prospective study of 100 women with CRD to assess the prevalence of gynaecological disorders in this group of patients. Assessment included the measurement of follicle stimulating hormone, luteinizing hormone, prolactin and oestradiol concentrations, cervical cytology and a pelvic ultrasound scan. We found that gynaecological problems are highly prevalent and frequently unrecognized. Of these women, 58% had a menstrual disorder, with uncontrolled menorrhagia being a significant problem when it aggravated the chronic anaemia of renal disease, and 35% were menopausal, including seven women under the age of 40 years. Menopausal symptoms were undertreated. We identified a 14-fold increase in premature ovarian failure secondary to CRD and the use of cyclophosphamide therapy. In all, 22% of the women were subfertile and 10% had an abnormal smear, with cervical dyskariosis being significantly increased because of long-term immunosuppression. Contraceptive advice had often been absent or inappropriate. We conclude that formal gynaecological review should be routinely available for women with CRD.


Chromonychia secondary to chemotherapy.
Chemotherapy drugs can affect the skin and its appendages. Several clinical presentations can be observed, depending on the affected structure. The most common dermatological side effect is chromonychia. The main causative agents are: (1) cyclophosphamide, which can provoke a diffuse, black pigmentation, longitudinal striae and dark grey pigmentation located proximally on the nails; (2) doxorubicin, which promotes dark brown bands alternating with white striae and dark brown pigmentation in transverse bands, and (3) hydroxyurea, which produces a distal, diffuse, dark brown pigmentation. In the majority of cases, the effects are reversible after the suspension of the causative agent for a few months. We report a patient who developed chromonychia while undergoing treatment with cyclophosphamide, vincristine, doxorubicin, dexamethasone, methotrexate and cytarabine for acute lymphocytic leukemia.


Homogeneous oxidation of aqueous solutions of atrazine and fenitrothion through dark and photo-Fenton reactions.
The homogeneous catalytic oxidation of atrazine and fenitrothion in single compound-containing solutions and in mixture, in pure water, was investigated in dark and light-driven oxidation systems using H(2)O(2) as the oxidant and Fe(II) as the catalyst. It was shown that the degradation during the dark Fenton reactions takes place during the first 2h and further prolongation of the reaction time does not lead to enhanced removals. The same holds true for the light-driven reactions. With the photo-Fenton reaction, fenitrothion was completely mineralized while the TOC contained in the atrazine solution was removed by 57%. Both compounds in their parent form were completely eliminated from the first 5 min of the oxidation both when being in the single-containing solutions and in their binary mixture. The oxidation of the TOC in the atrazine solution follows a two-stage second-order kinetic behavior which is attributed to the complex reaction pathways occurring during the oxidation while that of the TOC in the fenitrothion solutions follows a pseudo-first order degradation kinetic and that of binary mixture a two-stage degradation kinetics. The results obtained in this study, clearly indicate that the light-driven Fenton oxidation can be very efficient for the removal of atrazine and especially fenitrothion from polluted waters.


iDISCO: a simple, rapid method to immunolabel large tissue samples for volume imaging.
The visualization of molecularly labeled structures within large intact tissues in three dimensions is an area of intense focus. We describe a simple, rapid, and inexpensive method, iDISCO, that permits whole-mount immunolabeling with volume imaging of large cleared samples ranging from perinatal mouse embryos to adult organs, such as brains or kidneys. iDISCO is modeled on classical histology techniques, facilitating translation of section staining assays to intact tissues, as evidenced by compatibility with 28 antibodies to both endogenous antigens and transgenic reporters like GFP. When applied to degenerating neurons, iDISCO revealed unexpected variability in number of apoptotic neurons within individual sensory ganglia despite tight control of total number in all ganglia. It also permitted imaging of single degenerating axons in adult brain and the first visualization of cleaved Caspase-3 in degenerating embryonic sensory axons in vivo, even single axons. iDISCO enables facile volume imaging of immunolabeled structures in complex tissues. PAPERCLIP:


Relationship between physical activity and coping with stress in people with multiple sclerosis: A moderated mediation model with self-efficacy and disability level as variables.
An increasing number of studies support the beneficial relationship between physical activity and stress coping in people with multiple sclerosis (MS). However, there has been limited understanding of the variables that may influence the nature of this relationship. Therefore, based on the social-cognitive framework and previous research, we aimed to examine the association between the habitual physical activity of people with MS and their coping effectiveness. Furthermore, we sought to determine the extent to which self-efficacy acts as a mediator in this relationship, considering the level of disability as a moderator variable. In this cross-sectional study, a total of 351 people with MS participated. The participants were asked to complete several assessment tools, including the Mini-COPE Inventory for Measurement-Coping with Stress, the Generalized Self-Efficacy Scale, and the Godin Leisure-Time Exercise Questionnaire. Additionally, a neurologist assessed the severity of the disease using the Expanded Disability Status Scale. Information on the demographic and clinical characteristics of the participants was collected via a self-report survey. Two moderated mediation analyses were conducted as part of the study. The study findings indicated a positive correlation between engagement in physical activity and self-efficacy among participants with high and medium disability levels. This, in turn, demonstrated a positive association with effective stress-coping strategies and a negative association with ineffective coping methods. In particular, a significant relationship was observed between involvement in physical activity and self-efficacy in participants with high disability, while it was not statistically significant in participants with low disability. Physical activity was associated with improved psychosocial functioning in people with high levels of disability caused by MS. This association may be attributed to factors such as increased self-efficacy and improved stress coping. However, the relationship between physical activity and psychosocial functioning was less evident in people with low disability caused by MS.


Strain-specific difference that affects the inhibition of division of Escherichia coli filaments by chloramphenicol.
The Escherichia coli mutations ts1882 and ts2158 cause temperature-sensitive septum formation and result in growth of cells as long, multinucleate, nonseptate filaments at 42 C. When filaments are transferred to 28 C, they divide into short cells. Chloramphenicol, when added to cultures of filaments at the time of temperature reduction. Inhibited division of filaments when these temperature-sensitive mutations were present in the K-12 strain AB1157. However, when the ts1882 and ts2158 mutations were present in another k-12 strain, UTH4113, filaments of these strains divided in the presence of chloramphenicol.


In-Flight Hypoxemia in a Tracheostomy-Dependent Infant.
Millions of passengers board commercial flights every year. Healthcare providers are often called upon to treat other passengers during in-flight emergencies. The case presented involves an anesthesia resident treating a tracheostomy-dependent infant who developed hypoxemia on a domestic flight. The patient had an underlying congenital muscular disorder and was mechanically ventilated while at altitude. Although pressurized, cabin barometric pressure while at altitude is less than at sea level. Due to this environment patients with underlying pulmonary or cardiac pathology might not be able to tolerate commercial flight. The Federal Aviation Administration (FAA) has mandated a specific set of medical supplies be present on all domestic flights in addition to legislature protecting "Good Samaritan" providers.


The influence of the thymine C5 methyl group on spontaneous base pair breathing in DNA.
Sequences of four or more AT base pairs without a 5'-TA-3' step, so-called A-tracts, influence the global properties of DNA by causing curvature of the helix axis if phased with the helical repeat and also influence nucleosome packaging. Hence it is interesting to understand this phenomenon on the molecular level, and numerous studies have been devoted to investigations of dynamical and structural features of A-tract DNA. It was early observed that anomalously slow base pair-opening kinetics were a striking physical property unique to DNA A-tracts (Leroy, J. L., Charretier, E., Kochoyan, M., and Gueron, M. (1988) Biochemistry 27, 8894-8898). Furthermore, a strong correlation between DNA curvature and anomalously slow base pair-opening dynamics was found. In the present work it is shown, using imino proton exchange measurements by NMR spectroscopy that the main contribution to the dampening of the base pair-opening fluctuations in A-tracts comes from the C5 methylation of the thymine base. Because the methyl group has been shown to have a very limited effect on the DNA curvature as well as the structure of the DNA helix, the thymine C5 methyl group stabilizes the helix directly. Empirical potential energy calculations show that methylation of the tract improves the stacking energy of a base pair with its neighbors in the tract by 3-4 kcal/mol.


HIV infection: the spectrum of symptoms and disease in male and female patients attending a London hospice.
To delineate the demographic features and clinical profile of male and female individuals utilizing a respite, rehabilitation and hospice centre in London, a retrospective study of patient notes was undertaken. The subjects were 59 male and 59 female HIV patients at first admission to the hospice. The study revealed clinically important differences between men and women and also identified special problems for African women and intravenous drug users. Women experienced more constipation, headache and musculoskeletal pain than men. Men suffered more neuropathic pain and visual loss. Lower respiratory tract infections were more common in women, especially in those with a history of injection drug use. Gynaecological morbidity was common. There was no difference in AIDS diagnoses between men and women. HIV encephalopathy developed in 23% of the cohort during the follow-up period which has implications for provider units. There are increasing numbers of HIV-positive women and injection drug users being referred for palliative care services. Multidisciplinary teams should be aware of the particular symptom profile of these groups.


Auxiliary liver organ formation by implantation of spleen-encapsulated hepatocytes.
Hepatocyte transplantation is an attractive alternative to orthotopic liver transplantation. However, its application has been limited because of its short-term success only. Here we report a new approach to hepatocyte transplantation resulting in the generation of an auxiliary liver in vivo. Isolated primary hepatocytes were encapsulated in isolated spleens and then transplanted by attaching the spleens to the livers of recipient animals (mice or rats) using biodegradable adhesive. A vascular network was rapidly established, and protein molecules circulated freely between the transplanted spleen and the liver, to which they adhered. In contrast, the spleen, which did not adhere to the liver or adhered elsewhere (adipose tissue or peritoneum), did not become vascularized but shrank and died. Encapsulation of hepatocytes in an isolated spleen enhanced their survival significantly, and co-encapsulation of Engelbreth- Holm-Swarm gel together with the hepatocytes further enhanced it. The encapsulated hepatocytes expressed liver-specific differentiation genes for more than 3 weeks. Plasma albumin concentrations in Nagase analbuminemic rats began to increase 3 days after transplantation. The transplanted hepatic cells migrated into the liver parenchyma, whereas the spleen was absorbed. Thus, we have developed a novel, simple approach for the rapid and efficient formation of functional auxiliary liver using a modified hepatocyte transplantation method.


Imaging Evaluation of Pediatric Parotid Gland Abnormalities.
Parotid gland lesions in children can be divided into benign or malignant. The age of the patient helps narrow the differential diagnosis, with vascular and congenital lesions being more frequent in the 1st year of life, while solid tumors are more frequent in older children. Inflammatory disease usually has rapid onset in comparison with that of neoplastic or congenital processes, which have more gradual clinical evolution. Currently, multiple imaging techniques are available to study the parotid region, such as US, CT, and MRI. However, it is still a challenge to distinguish nonmalignant lesions from malignant ones. US is the first-line diagnostic approach in children to characterize the morphology and vascularity of these lesions. CT in children may be indicated for evaluation of abscesses or sialolithiasis. MRI is the imaging modality of choice for investigating the nature of the lesion and its extent. In addition to complete and detailed clinical information, knowledge of parotid gland anatomy and characteristic radiologic features of parotid disorders is essential for optimal radiologic evaluation and avoiding unnecessary interventional diagnostic procedures or treatment. This article illustrates a variety of entities (congenital, inflammatory, vascular, neoplastic) that can occur in the parotid gland, highlighting the most frequent radiologic patterns of manifestation and correlating them with clinical, surgical, and pathologic findings. ©RSNA, 2018.


Surviving entanglement in optic-microwave conversion by an electro-optomechanical system.
In recent development of quantum technologies, a frequency conversion of quantum signals has been studied widely. We investigate the optic-microwave entanglement that is generated by applying an electro-optomechanical frequency conversion scheme to one mode in an optical two-mode squeezed vacuum state. We quantify entanglement of the converted two-mode Gaussian state, where surviving entanglement of the state is analyzed with respect to the parameters of the electro-optomechanical system. Furthermore, we show that there exists an upper bound for the entanglement that survives after the conversion of highly entangled optical states. Our study provides a theoretical platform for a practical quantum illumination system.


Three-dimensional optical sectioning by scanning confocal electron microscopy with a stage-scanning system.
We evaluated the depth resolution of annular dark-field (ADF) scanning confocal electron microscopy (SCEM) with a stage-scanning system by observation of nanoparticles. ADF-SCEM is a three-dimensional (3D) imaging technique that we recently proposed. An ADF-SCEM instrument involves a pinhole aperture before a detector for rejecting electrons from the out-of-focal plane in a specimen and an annular aperture under the specimen for collecting only scattered electrons. The stage-scanning system enables us to directly obtain optical slice images perpendicular and parallel to an optical axis at a desired position. In particular, the parallel slices visualize the elongation of nanoparticles along the optical axis, which depends on the depth resolution. ADF-SCEM effectively reduced the elongation length of the nanoparticles sufficiently to demonstrate depth sectioning, in comparison with scanning transmission electron microscopy and bright-field SCEM. The experimentally obtained length was nearly equal to the theoretically estimated one from the probe size considering the experimental conditions. Furthermore, we applied this ADF-SCEM technique to analysis of the 3D position of catalytic nanoparticles on carbon nanostructures.


Effects of restriction of sodium or administration of fludrocortisone on parotid salivary kallikrein in man.
Urinary kallikrein is increased by restriction of dietary sodium and by administration of fludrocortisone, a sodium-retaining steroid. In order to determine whether salivary kallikrein responds similarly, we studied 16 normal volunteers after 1-week periods of daily intake of 9, 109, and 259 mEQ of sodium; 10 subjects were studied after addition of 0.6 mg/day fludrocortisone for a week to a regimen of 109 mEq/day sodium. During sodium restriction, parotid saliva had a significantly higher mean concentration of kallikrein ad potassium and a significantly lower concentration of sodium than during periods of intake of 109 or 259 mEq/day sodium. Sodium restriction also caused significantly higher urinary excretion of kallikrein and aldosterone. Salivary amylase remained unchanged during the three sodium periods. Administration of fludrocortisone significantly increased the mean concentration of parotid kallikrein and excretion of urinary kallikrein in comparison with control levels, however the concentrations of parotid sodium and potassium did not change significantly. Four patients studied before and after removal of aldosterone-producing adenomas each showed decreased concentrations of parotid kallikrein and potassium and increased concentrations of parotid sodium after surgery. It is concluded that both salivary and urinary kallikrein increased in response to restriction of sodium and that these increases were mediated by levels of sodium-retaining steroid. Increased output of kallikrein in response to increased levels of sodium-retaining steroid may be a generalized response of organs that contain glandular kallikrein and can conserve sodium.


Structural and biochemical properties of bidentate tetraaquarhodium(III) complexes of inorganic pyrophosphate and adenosine 5'-diphosphate.
The structural and biochemical properties of the alpha,beta-bidentate tetraaquarhodium(III) complexes of inorganic pyrophosphate [Rh(H2O)4PP] and adenosine diphosphate [Rh(H2O)4ADP] are examined. These Rh(III) complexes are exchange-inert analogues of the corresponding physiologically important MgIIPP and MgIIADP complexes. The crystal structure of [Rh(H2O)4H2P2O7]+Cl- shows that the six-membered chelate ring adopts a twist-boat conformation with an unusually high puckering amplitude of 0.756 (3) A. The Rh coordination distances average 2.02 (1) A, while the bridge P-O bonds are virtually equal in length. All 10 protons of the complex participate in hydrogen bonding. There are two intramolecular hydrogen bonds between the phosphate oxygen atoms and the axially coordinated water molecules. The Rh(H2O)4PP complex was found to be a substrate for yeast inorganic pyrophosphatase, with Ki = 0.063 (7) mM and Vm = 500 (100) min-1. The two screw sense isomers of Rh(H2O)4ADP were prepared from (Rp)-[alpha-16O,18O]ADP and assigned configuration on the basis of the magnitude of their 31P NMR isotopic chemical shifts. The Rh(H2O)4ADP complex binds a number of kinases as tightly as MgADP. Arginine kinase and creatine kinase were shown to bind the delta Rh(H2O)4ADP isomer 7 and 45 times tighter, respectively, than the lambda isomer. The reactivity of Rh(H2O)4PP with pyrophosphatase is comparable to that of Cr(H2O)4PP, and the binding affinities of the Rh(H2O)4ADP screw sense isomers for kinases are also comparable to those observed for the corresponding Cr(H2O)4ADP screw sense isomers.


N-Type Quinoidal Polymers Based on Dipyrrolopyrazinedione for Application in All-Polymer Solar Cells.
Conjugated molecules and polymers with intrinsic quinoidal structure are promising n-type organic semiconductors, which have been reported for application in field-effect transistors and thermoelectric devices. In principle, the molecular and electronic characteristics of quinoidal polymers can also enable their application in organic solar cells. Herein, two quinoidal polymers, named PzDP-T and PzDP-ffT, based on dipyrrolopyrazinedione were synthesized and used as electron acceptors in all-polymer solar cells (all-PSCs). Both PzDP-T and PzDP-ffT showed suitable energy levels and wide light absorption range that extended to the near-infrared region. When combined with the polymer donor PBDB-T, the resulting all-PSCs based on PzDP-T and PzDP-ffT exhibited a power conversion efficiency (PCE) of 1.33 and 2.37 %, respectively. This is the first report on the application of intrinsic quinoidal conjugated polymers in all-PSCs. The photovoltaic performance of the all-PSCs was revealed to be mainly limited by the relatively poor and imbalanced charge transport, considerable charge recombination. Detailed investigations on the structure-performance relationship suggested that synergistic optimization of light absorption, energy levels, and charge transport properties is needed to achieve more successful application of intrinsic quinoidal conjugated polymers in all-PSCs.


* Gelatin Scaffolds Containing Partially Sulfated Cellulose Promote Mesenchymal Stem Cell Chondrogenesis.
Articular cartilage has a limited capacity to heal after damage from injury or degenerative disease. Tissue engineering constructs that more closely mimic the native cartilage microenvironment can be utilized to promote repair. Glycosaminoglycans (GAGs), a major component of the cartilage extracellular matrix, have the ability to sequester growth factors due to their level and spatial distribution of sulfate groups. This study evaluated the use of a GAG mimetic, cellulose sulfate, as a scaffolding material for cartilage tissue engineering. Cellulose sulfate can be synthesized to have a similar level and spatial distribution of sulfates as chondroitin sulfate C (CSC), the naturally occurring GAG. This partially sulfated cellulose (pSC) was incorporated into a fibrous gelatin construct by the electrospinning process. Scaffolds were characterized for fiber morphology and overall stability over time in an aqueous environment, growth factor interaction, and for supporting mesenchymal stem cell (MSC) chondrogenesis in vitro. All scaffold groups had micron-sized fibers and maintained overall stability in aqueous environments. Increasing concentrations of the transforming growth factor-beta 3 (TGF-β3) were detected on scaffolds with increasing pSC. MSC chondrogenesis was enhanced on the scaffold with the highest pSC concentration as seen with the highest collagen type II production, collagen type II immunostaining, expression of cartilage-specific genes, and ratio of collagen type II to collagen type I production. These studies demonstrated the potential of pSC sulfate as a scaffolding material for cartilage tissue engineering.


Weighted gene co-expression network analysis identifies dysregulated B-cell receptor signaling pathway and novel genes in pulmonary arterial hypertension.
Pulmonary arterial hypertension (PAH) is a devastating cardio-pulmonary vascular disease in which chronic elevated pulmonary arterial pressure and pulmonary vascular remodeling lead to right ventricular failure and premature death. However, the exact molecular mechanism causing PAH remains unclear. RNA sequencing was used to analyze the transcriptional profiling of controls and rats treated with monocrotaline (MCT) for 1, 2, 3, and 4 weeks. Weighted gene co-expression network analysis (WGCNA) was employed to identify the key modules associated with the severity of PAH. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to explore the potential biological processes and pathways of key modules. Real-time PCR and western blot analysis were used to validate the gene expression. The hub genes were validated by an independent dataset obtained from the Gene Expression Omnibus database. A total of 26 gene modules were identified by WGCNA. Of these modules, two modules showed the highest correlation with the severity of PAH and were recognized as the key modules. GO analysis of key modules showed the dysregulated inflammation and immunity, particularly B-cell-mediated humoral immunity in MCT-induced PAH. KEGG pathway analysis showed the significant enrichment of the B-cell receptor signaling pathway in the key modules. Pathview analysis revealed the dysregulation of the B-cell receptor signaling pathway in detail. Moreover, a series of humoral immune response-associated genes, such as BTK, BAFFR, and TNFSF4, were found to be differentially expressed in PAH. Additionally, five genes, including BANK1, FOXF1, TLE1, CLEC4A1, and CLEC4A3, were identified and validated as the hub genes. This study identified the dysregulated B-cell receptor signaling pathway, as well as novel genes associated with humoral immune response in MCT-induced PAH, thereby providing a novel insight into the molecular mechanisms underlying inflammation and immunity and therapeutic targets for PAH.


Racial and ethnic disparities in cancer survival by neighborhood socioeconomic status in Surveillance, Epidemiology, and End Results (SEER) Registries.
Reducing cancer disparities is a major public health objective. Disparities often are discussed in terms of either race and ethnicity or socioeconomic status (SES), without examining interactions between these variables. Surveillance, Epidemiology, and End Results (SEER)-18 data, excluding Alaska Native and Louisiana registries, from 2002 to 2008, were used to estimate five-year, cause-specific survival by race/ethnicity and census tract SES. Differences in survival between groups were used to assess absolute disparities. Hazard ratios were examined as a measure of relative disparity. Interactions between race/ethnicity and neighborhood SES were evaluated using proportional hazard models. Survival increased with higher SES for all racial/ethnic groups and generally was higher among non-Hispanic white and Asian/Pacific Islander (API) than non-Hispanic black and Hispanic cases. Absolute disparity in breast cancer survival among non-Hispanic black vs non-Hispanic white cases was slightly larger in low-SES areas than in high-SES areas (7.1% and 6.8%, respectively). In contrast, after adjusting for stage, age, and treatment, risk of mortality among non-Hispanic black cases compared with non-Hispanic white cases was 21% higher in low-SES areas and 64% higher in high-SES areas. Similarly, patterns of absolute and relative disparity compared with non-Hispanic whites differed by SES for Hispanic breast cancer, non-Hispanic black colorectal cancer, and prostate cancer cases. Statistically significant interactions existed between race/ethnicity and SES for colorectal and female breast cancers. In health disparities research, both relative and absolute measures provide context. A better understanding of the interactions between race/ethnicity and SES may be useful in directing screening and treatment resources toward at-risk populations.


A method to study human synaptic protein-protein interactions by using flow cytometry coupled to proximity ligation assay (Syn-FlowPLA).
Synapses are highly specialized sites characterized by intricate networks of protein-protein interactions (PPIs) important to maintain healthy synapses. Therefore, mapping these networks could address unsolved questions about human cognition, synaptic plasticity, learning, and memory in physiological and pathological conditions. The limitation of analyzing synaptic interactions in living humans has led to the development of methods to isolate synaptic terminals (synaptosomes) from cryopreserved human brains. Here, we established a method to detect synaptic PPIs by applying flow cytometric proximity ligation assay (FlowPLA) to synaptosomes isolated from frozen human frontal cortex (FC) and hippocampus (HP) (Syn-FlowPLA). Applying this method in synaptosomes, we were able to detect the known post-synaptic interactions between distinct subtypes of N-methyl-D-aspartate glutamate receptors (NMDARs) and their anchoring postsynaptic density 95 protein (PSD95). Moreover, we detected the known pre-synaptic interactions between the SNARE complex proteins synaptosomal-associated protein of 25 kDa (SNAP25), synaptobrevin (VAMP2), and syntaxin 1a (STX1A). As a negative control, we analyzed the interaction between mitochondrial superoxide dismutase 2 (SOD2) and PSD95, which are not expected to be physically associated. PPIs have been studied in vitro primarily by co-immunoprecipitation, affinity chromatography, protein-fragment complementation assays (PCAs), and flow cytometry. All these are valid approaches; however, they require more steps or combination with other techniques. PLA technology identifies PPIs with high specificity and sensitivity. The Syn-FlowPLA described here allows rapid analyses of PPIs, specifically within the synaptic compartment isolated from frozen autopsy specimens, achieving greater target sensitivity. Syn-FlowPLA, as presented here, is therefore a useful method to study human synaptic PPI in physiological and pathological conditions.


ATGL has a key role in lipid droplet/adiposome degradation in mammalian cells.
Lipid droplets (LDs), also called adiposomes, are found in many eukaryotic cells, and are highly upregulated in lipid-storage cells, such as adipocytes. The mechanism by which adiposomes and their component neutral lipids are degraded is an important health issue with the rapidly spreading epidemic of obesity. Recently, a novel triglyceride lipase (adipose triglyceride lipase (ATGL)) that catalyses the initial step in triglyceride hydrolysis in adipocyte LDs was identified. Here, we show that ATGL also functions in non-adipocyte cells, and has an important role in LD degradation in these cells. Overexpression of wild-type ATGL causes a marked decrease in LD size, whereas a catalytically inactive mutant retains the ability to localize to LDs, but is unable to decrease their size. Depletion of ATGL by RNA interference leads to a significant increase in the size of LDs. These results show that ATGL has an important role in LD/adiposome turnover in mammalian cells.


Mechanism of action of cinodine, a glycocinnamoylspermidine antibiotic.
The mechanism of action of cinodine, a glycocinnamoylspermidine antibiotic, was investigated. Upon addition of cinodine to growing cultures of Escherichia coli, a rapid decline in viable cell numbers was observed. Culture turbidity continued to increase for a short period before plateauing. Microscopic examination indicated that the antibiotic-treated cells continued to elongate with subsequent formation of serpentine-like structures. Radioisotopic-labeling studies of E. coli demonstrated that deoxyribonucleic acid (DNA) synthesis was immediately and irreversibly inhibited upon addition of cinodine. Ribonucleic acid synthesis was reduced after a significant delay, whereas protein synthesis remained unaffected. There was a minor degree of inhibition of incorporation of radiolabeled diaminopimelic acid into cell wall material. Cinodine likewise inhibited bacteriophage T7 DNA synthesis in infected E. coli cells. After inhibition of E. coli DNA synthesis by cinodine, intracellular DNA degradation was observed. Equilibrium dialysis studies demonstrated that the drug physically bound to DNA. These data indicate that cinodine functions as a potent irreversible inhibitor of bacterial and phage DNA synthesis.


Effects of perlite and caustic soda on microorganism activities of leachate in a sequence batch reactor.
The purpose of this article is to evaluate the effect of adsorbents and alkali pre-treatment on microorganism activities of activated sludge (AS) for the treatment of landfill leachate (LFL). The chemical oxygen demand (COD) and BOD5/COD ratio of LFL used in this research were 10,500 and 0.68, respectively. In order to survey the role of porous absorbent, perlite was employed as an alternative with low porosity and was compared to powdered activated carbon (PAC), which has been most widely used in the treatment process. As a result, the COD removal efficiency increased from 32% to 47.7% when alkali LFL was loaded to the sequence batch reactors (SBRs) at the optimum conditions of the biological process. Also, at the same condition, both SBRs containing PAC and perlite showed COD removals of over 81% and 72%, respectively. The specific oxygen uptake rate (SOUR) showed that alkali pre-treatment reduces the toxicity effect of heavy metals on microorganism activities. The adsorption capacity (the uptake of COD) was analyzed by Langmuir and Freundlich isotherm models. Further, the kinetic study of COD adsorption during the treatment process demonstrated that the alkali pre-treatment of LFL proceeded faster and was intensified by the presence of adsorbents.


Developmental trajectories of infants born at less than 30 weeks' gestation on the Bayley-III Scales.
To describe the cognitive, language and motor developmental trajectories of children born very preterm and to identify perinatal factors that predict the trajectories. Data from a cohort of 1142 infants born at <30 weeks' gestation who were prospectively assessed on the Bayley Scales of Infant and Toddler Development, third edition (Bayley-III) at 3, 6, 12 and 24 months corrected age, were analysed using the Super Imposition by Translation and Rotation (SITAR) growth curve analysis model. Developmental trajectory SITAR models for Bayley-III cognitive, language (receptive and expressive communication subscales) and motor (fine and gross motor subscales) scores. The successfully fitted SITAR models explained 62% of variance in cognitive development, 68% in receptive communication, 53% in fine motor and 68% in the gross motor development. There was too much variation in the expressive communication subscale to fit a SITAR model. The rate of development (gradient of the curve) best explains the variation in trajectories of development in all domains. Lower gestational age, lower birth weight and male sex significantly predicted a slower rate of development. The rate of development, rather than single time point developmental assessment, best predicts the very preterm infant's developmental trajectory and should be the focus for monitoring and early intervention.


Helmets for snow sports: prevalence, trends, predictors and attitudes to use.
Traumatic head injury is the leading cause of mortality and serious morbidity in snow sports and is increasing in incidence. Helmet use in snow sports can reduce the incidence of head injury by up to 60%. Mandatory helmet use is not legislated in most recreational settings for snow sports. This study aimed to quantify the prevalence and trends of helmet use by skiers and snowboarders as well as to investigate predictors of helmet use and reasons why they are worn. A cross-sectional study including observation of skiers and snowboarders allocated to adult and child groups (5,267 persons), retrospective review of ski patrol accident report forms recorded between 2003 and 2008 (3,984), and completion of specially designed questionnaires by randomly approached snow sport participants (1,029). In 2008, 16% of adults and 67% of children wore helmets. Helmet use increased for adult and child participants between 2003 and 2008. Children, men, and snowboarders were significantly more likely to wear a helmet than their respective counterparts. Significant predictors of helmet use were level of experience, past major crash requiring medical assistance, snow sport lessons, and location of activity. Key reasons for helmet use and disuse were identified. Child helmet use far exceeded adult helmet use. Rates of voluntary helmet use are increasing but many remain unprotected from the benefits of a helmet. Identification of the snow sport participants least likely to wear a helmet and their reasons for not doing so should allow targeted helmet promotion and injury prevention strategies to reduce serious head injuries in snow sports.


Contributions of KRAS and RAL in non-small-cell lung cancer growth and progression.
KRAS mutations are poor prognostic markers for patients with non-small-cell lung cancer (NSCLC). RALA and RALB GTPases lie downstream of RAS and are implicated in RAS-mediated tumorigenesis. However, their biological or prognostic role in the context of KRAS mutation in NSCLC is unclear. Using expression analysis of human tumors and a panel of cell lines coupled with functional in vivo and in vitro experiments, we evaluated the prognostic and functional importance of RAL in NSCLC and their relationship to KRAS expression and mutation. Immunohistochemical (N = 189) and transcriptomic (N = 337) analyses of NSCLC patients revealed high RALA and RALB expression was associated with poor survival. In a panel of 14 human NSCLC cell lines, RALA and RALB had higher expression in KRAS mutant cell lines whereas RALA but not RALB activity was higher in KRAS mutant cell lines. Depletion of RAL paralogs identified cell lines that are dependent on RAL expression for proliferation and anchorage independent growth. Overall, growth of NSCLC cell lines that carry a glycine to cystine KRAS mutation were more sensitive to RAL depletion than those with wild-type KRAS. The use of gene expression and outcome data from 337 human tumors in RAL-KRAS interaction analysis revealed that KRAS and RAL paralog expression jointly impact patient prognosis. RAL GTPase expression carries important additional prognostic information to KRAS status in NSCLC patients. Simultaneously targeting RAL may provide a novel therapeutic approach in NSCLC patients harboring glycine to cystine KRAS mutations.


The development of symbol-infused joint engagement.
Fifty-six children were observed longitudinally from 18 to 30 months of age interacting with their mothers during a Communication Play that contained 8 scenes designed to encourage interacting, requesting, commenting, and narrating. Of primary concern was how often symbols infused the child's states of engagement with people and objects and how experience in such symbol-infused states related to language acquisition. Findings indicate that symbols increasingly infuse joint engagement, and that both the timing and the trajectory vary widely among typically developing toddlers, especially during the last half of the 2nd year. Moreover, variations in amount of symbol-infused supported joint engagement may both be influenced by variations in the onset of language and contribute to differences in language facility at 30 months.


A systematic review of group therapy programs for smoking cessation in Asian countries.
Tobacco causes more than 8 million deaths each year. Behavioral interventions such as group therapy, which provides counselling for smoking cessation, can be delivered in group form and smokers who receive cessation counselling are more likely to quit smoking compared to no assistance. We review the evidence of group-based counselling for smoking cessation for smokers in Asian countries. The review aims to determine the availability of group-based therapy for smoking cessation in Asian countries. The outcome measured was abstinence from smoking following group therapy. Electronic database searches in PubMed, OVID Medline, SCOPUS, Google Scholar, and PsycINFO, using keywords such as: 'smoking', 'cigarette', 'tobacco', 'nicotine', 'group therapy' and 'cessation' (smok*, *cigarette*, tobacco, nicotine, group therap*, cessation) were used. The results were reported following PRISMA and PROSPERO guidelines. Review Manager was used for data analysis. A total of 21251 records were retrieved for screening the abstracts. In all, 300 articles for review were identified and assessed for eligibility. Nine articles, including Cochrane reviews, randomized control trials, cohort, observational and cross-sectional studies, were included in the final review. There were three observational qualitative studies, two prospective cohort studies, two crosssectional studies, one non-randomized quasi-experimental study and a single cluster-randomized, controlled trial. Group therapy was found to significantly increase the abstinence rate. Group therapy provided at the workplace, smoking cessation services, availability of pharmacotherapy, and socioeconomic status, appear to be key factors determining success. Evidence of the use of group therapy for smoking cessation in Asian countries is still lacking despite publications in the Western population showed that group therapy was effective. Further research on group-based interventions for smoking cessation in Asian countries is required and direct one-to-one comparisons between group therapy and individual therapy for smokers who want to quit smoking, are needed.


The anaphase promoting complex induces substrate degradation during neuronal differentiation.
The anaphase promoting complex (APC) is an E3 ubiquitin ligase required for the metaphase-to-anaphase transition and mitotic exit. However, APC also plays roles in G(1), where it is regulated by Cdh1, and APC activity has also been detected in differentiated and non-proliferating cells, suggesting that it may play roles outside the cell cycle. Here, we report that disrupting APC(Cdh1) activity inhibits neurite outgrowth of both PC12 pheochromocytoma cells and primary cerebellar granule cells. APC(Cdh1) activity dramatically increases as PC12 cells differentiate in response to nerve growth factor. Furthermore, a key target degraded by APC(Cdh1) following nerve growth factor treatment is the F-box protein Skp2, and APC(Cdh1)-mediated destruction of Skp2 is essential for proper terminal differentiation of neuronal precursors.


Transcatheter mitral valve repair for functional mitral regurgitation: Evaluating the evidence.
Two trials (Cardiovascular Outcomes Assessment of the MitraClip Percutaneous Therapy for Heart Failure Patients with Functional Mitral Regurgitation Trial and Percutaneous Repair with the MitraClip Device for Severe Functional/Secondary Mitral Regurgitation Trial) were published in 2018 evaluating the effectiveness and safety of transcatheter repair for patients with heart failure with significant functional mitral regurgitation, which yielded different results. This article reviews the strength of the evidence, differences in trial designs, ethical and implementation implications, and delineates future research needs to help guide the appropriate dissemination of transcatheter repair for functional patients with mitral regurgitation. The National Heart, Lung, and Blood Institute convened a workshop of interdisciplinary experts to address these objectives. Transcatheter repair of functional mitral regurgitation can provide significant benefits in terms of heart failure hospitalizations, survival, and quality of life when appropriate heart failure candidates with moderate to severe or severe mitral regurgitation while on optimal guideline-directed medical therapy can be identified. Key ingredients for success are preoperative evaluation and management and postoperative care by an interdisciplinary heart team. Given the discordance observed between trials, ongoing innovation in patient management, and potential expansion of indications for use, the evidence base must be expanded to optimize appropriate implementation of this complex therapy. This will require more complete capture of outcome data in real-world settings for all eligible candidates whether or not they receive this therapy. Inevitably, the indications for use of this therapy will expand, as will the devices and therapeutic approaches for this population, necessitating the study of comparative effectiveness through randomized trials or observational studies. Moreover, given the substantial variations in care delivery, conducting implementation research to delineate characteristics of the optimal care model would be of benefit.


Comprehension demands modulate re-reading, but not first pass reading behavior.
Several studies have examined effects of explicit task demands on eye movements in reading. However, there is relatively little prior research investigating the influence of implicit processing demands. In the present study, processing demands were manipulated by means of a between-subject manipulation of comprehension question difficulty. Consistent with previous results from Wotschack and Kliegl (2013), the question difficulty manipulation influenced the probability of regressing from late in sentences and re-reading earlier regions; readers who expected difficult comprehension questions were more likely to re-read. However, this manipulation had no reliable influence on eye movements during first pass reading of earlier sentence regions. Moreover, for the subset of sentences that contained a plausibility manipulation, the disruption induced by implausibility was not modulated by the question manipulation. We interpret these results as suggesting that comprehension demands influence reading behavior primarily by modulating a criterion for comprehension that readers apply after completing first-pass processing.


Refractive index sensing with subradiant modes: a framework to reduce losses in plasmonic nanostructures.
Plasmonic modes with long radiative lifetimes, subradiant modes, combine strong confinement of the electromagnetic energy at the nanoscale with a steep spectral dispersion, which makes them promising for biochemical sensors or immunoassays. Subradiant modes have three decay channels: Ohmic losses, their extrinsic coupling to radiation, and possibly their intrinsic dipole moment. In this work, the performance of subradiant modes for refractive index sensing is studied with a general analytical and numerical approach. We introduce a model for the impact that has different decay channels of subradiant modes on the spectral resolution and contrast. It is shown analytically and verified numerically that there exists an optimal value of the mode coupling for which the spectral dispersion of the resonance line shape is maximal. The intrinsic width of subradiant modes determines the value of the dispersion maximum and depends on the penetration of the electric field in the metallic nanostructure. A figure of merit, given by the ratio of the sensitivity to the intrinsic width, which are both intrinsic properties of subradiant modes, is introduced. This figure of merit can be directly calculated from the line shape in the far-field optical spectrum and accounts for the fact that both the spectral resolution and contrast determine the limit of detection. An expression for the intrinsic width of a plasmonic mode is derived and calculated from the line shape parameters and using perturbation theory. The method of analysis introduced in this work is illustrated for dolmen and heptamer nanostructures. Fano-resonant systems have the potential to act as very efficient refractive index sensing platforms compared to Lorentz-resonant systems, due to control of their radiative losses. This study paves the way toward sensitive nanoscale biochemical sensors and immunoassays with a low limit of detection and, in general, any nano-optical device where Ohmic losses limit the performance.


Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers.
The presence of multipotent human limbal stromal cells resembling mesenchymal stromal cells (MSC) provides new insights to the characteristic of these cells and its therapeutic potential. However, little is known about the expression of stage-specific embryonic antigen 4 (SSEA-4) and the embryonic stem cell (ESC)-like properties of these cells. We studied the expression of SSEA-4 surface protein and the various ESC and MSC markers in the ex vivo cultured limbal stromal cells. The phenotypes and multipotent differentiation potential of these cells were also evaluated. Limbal stromal cells were derived from corneoscleral rims. The SSEA-4(+) and SSEA-4(-) limbal stromal cells were sorted by fluorescence-activated cells sorting (FACS). Isolated cells were expanded and reanalyzed for their expression of SSEA-4. Expression of MSC and ESC markers on these cells were also analyzed by FACS. In addition, expression of limbal epithelial and corneal stromal proteins such as ATP-binding cassette sub-family G member 2 (ABCG2), tumour protein p63 (p63), paired box 6 (Pax6), cytokeratin 3 (AE5), cytokeratin 10, and keratocan sulfate were evaluated either by immunofluorecence staining or reverse transcription polymerase chain reaction. Appropriate induction medium was used to differentiate these cells into adipocytes, osteocytes, and chondrocytes. Expanded limbal stromal cells expressed the majority of mesenchymal markers. These cells were negative for ABCG2, p63, Pax6, AE-5, and keratocan sulfate. After passaged, a subpopulation of these cells showed low expression of SSEA-4 but were negative for other important ESC surface markers such as Tra-1-60, Tra-1-81, and transcription factors like octamer-binding transcription factor 4 (Oct4), SRY(sex determining region Y)-box 2 (Sox2), and Nanog. Early passaged cells when induced were able to differentiate into adipocytes, osteocytes and chondrocytes. The expanded limbal stromal cells showed features of multipotent MSC. Our study confirmed the expression of SSEA-4 by a subpopulation of cultured limbal stromal cells. However, despite the expression of SSEA-4, these cells did not express any other markers of ESC. Therefore, we conclude that the cells did not show properties of ESC.


Nanostructured fat crystal systems.
A new understanding of the nature and organization of fat crystalline supramolecular structure, in particular at the nanoscale, has arisen in the past three years. These new findings have helped establish that the first step in the formation of a triacylglycerol network is the creation of nanocrystalline platelets that aggregate into polycrystalline clusters in the micrometer range, ultimately forming a three-dimensional network. This review explains how fat nanostructure can be characterized and highlights recent findings on how crystallization parameters influence the formation of fat nanocrystals. For instance, shear has been shown to modify not only nanoplatelet size but also their aggregation, affecting some macroscopic properties such as porosity and, therefore, the ability of the network to effectively bind liquid oil. This new information on fat nanostructure is relevant from scientific and technological standpoints and has opened up the possibility of nanoengineering material properties as well as developing new products and processes.


Plasma miR-200b in ovarian carcinoma patients: distinct pattern of pre/post-treatment variation compared to CA-125 and potential for prediction of progression-free survival.
Ovarian carcinomas (OvCa) are highly heterogeneous malignancies. We investigated four circulating plasma microRNAs (miR-21, miR-34a, miR-200b and miR-205) as candidate biomarkers. Using qPCR, we assessed the plasma concentration of these markers in 101 women, including 51 previously untreated OvCa patients, 25 healthy women and 25 patients bearing benign pelvic lesions. For a subset of 33 OvCa patients, the assay was repeated at the end of the primary treatment. The pattern of variations (post- minus pre-treatment) of concentration was compared to that of CA-125. A Cox regression model was used to study the association between variations and the progression-free survival (PFS). Plasma miR-200b proved to have a greater average concentration in OvCa samples (median 2-ΔΔCt = 15.18) than in samples linked to non-malignant lesions (median 2-ΔΔCt = 1.26, p-value = 0.0004). Its concentration was highly heterogeneous among OvCa patients, without any correlations with the FIGO stage and the pre-treatment CA-125 level. The decrease in CA-125 concentration was constant and often dramatic, while the variations of miR-200b concentration were much more diverse. The variation of miR-200b was marginally associated with the PFS (hazard ratio=2.95 95%CI=[0.94; 9.28], p=0.06) while miR-200b as a continuous time-dependent variable was significantly associated (HR=1.06 [1.02; 1.10], p=0.003). This study is the first direct empirical evidence that miR-200b can provide additional information, independent of CA-125 in OvCa patients.


A Form 990 Schedule H conundrum: how much of your bad debt might be charity?
IRS Form 990 Schedule H requires hospitals to estimate the amount of bad debt expense attributable to patients eligible for charity under the hospital's charity care policy. Responses to Schedule H, Part III.A.3 open up the entire patient collection process to examination by the IRS, state officials, and the public. Using predictive analytics can help hospitals efficiently identify charity-eligible patients when answering Part III.A.3.


Circulating and tumor-infiltrating myeloid-derived suppressor cells in patients with colorectal carcinoma.
Myeloid-derived suppressor cells (MDSCs) are a heterogeneous family of myeloid cells that suppress T cell immunity in tumor-bearing hosts. In patients with colon cancer, MDSCs have recently been described as Lin(-/low)HLA-DR(-)CD11b(+)CD33(+) cells correlating with cancer stage, metastasis and chemotherapy response. To learn in more detail the dynamic change and clinical relevance of circulating and tumor-infiltrating Lin(-/low)HLA-DR(-)CD11b(+)CD33(+) MDSC in colorectal cancer, we harvested the blood from 64 patients with varying stage of colorectal cancer and tumor and matched paraneoplastic tissues from 5 patients with advanced colorectal cancer, subjected them to multicolor flow cytometric analysis of percentage, absolute number and phenotype of MDSC and finally characterized their immunosuppressive functions. Our results demonstrate that peripheral blood from colorectal cancer patients contains markedly increased percentage and absolute number of Lin(-/low)HLA-DR(-)CD11b(+)CD33(+) MDSCs compared with healthy individuals, and this increase is closely correlated with clinical cancer stage and tumor metastasis but not primary tumor size and serum concentrations of cancer biomarker. A similar increase of MDSCs was also observed in the tumor tissues. Phenotyping MDSCs shows that they express high CD13 and CD39, low CD115, CD117, CD124 and PD-L1, and devoid of CD14, CD15 and CD66b, reminiscent of precursor myeloid cells. MDSCs from cancer patients but not healthy donors have the immunosuppressive activity and were able to inhibit in vitro autologous T-cell proliferation. Collectively, this study substantiates the presence of increased immunosuppressive circulating and tumor-resident Lin(-/low)HLA-DR(-)CD11b(+)CD33(+) MDSCs in patients with colorectal cancers correlating with cancer stage and metastasis, and suggests that pharmacologic blockade of MDSCs should be considered in future clinical trials.


Granulocyte-macrophage colony-stimulating factor regulation in murine T cells and its relation to cyclosporin A.
The coordinated expression of the hematopoietic growth factors, interleukin 3 (IL3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in activated T cells suggests common synthetic pathways. However, cyclosporin A (CsA) appears to differentially effect the synthesis of these two lymphokines. When the supernatants from the concanavalin A-stimulated thymoma EL-4 or the T-cell hybrid 2B4 were assayed on the GM-CSF/IL3-dependent PT-18 and on the IL3-dependent DA-1 cell lines, IL3 activity could not be detected following CsA treatment, but substantial growth activity, which was identified as GM-CSF, was observed on the PT-18 cell line. CsA did not affect the kinetics of GM-CSF release, but inhibited the release of IL3 over a period of 40 h after the cells were stimulated and treated with CsA. In addition, CsA could be added up to 1 h after stimulation of the cells without affecting GM-CSF activity but inhibiting completely the IL3 activity. To substantiate the inability of CsA to inhibit GM-CSF activity, GM-CSF gene expression was evaluated. By Northern analysis GM-CSF mRNA was not inhibited by doses of CsA up to 1 microgram/ml. The data reveal that CsA can dissociate between the production of IL3 and GM-CSF, suggesting that these two CSFs are regulated by different mechanisms.


[Clinical laboratory tests in diabetes mellitus].
The clinical diagnosis of diabetes is often prompted by symptoms such as increased thirst and urine volume, and weight loss. High levels of glucosuria are usually present. A single blood glucose estimation of diagnostic value indicated > or = 200 mg/dl at random, or > or = 140 mg/dl at fasting in specimens of venous plasma. Only if blood glucose values lie in the uncertain range (i.e., between the level that establish or exclude diabetes) need an oral glucose tolerance test (OGTT) be considered in order to establish the diagnosis status. Diagnostic interpretation of the 75 g OGTT responses is performed by the criteria of the expert committee of the Japan Diabetic Society. Glycated hemoglobin (HbA1c, HbA1) is widely used as a cumulative estimate of the mean blood glucose concentration over the preceding one approximately two months. Reference ranges of HbA1c are 4 approximately 6%. Labile glycohemoglobin often influences the estimate. Fructosamine and glycated albumin are also used as means of evaluating the degree of control. These data reveal the mean blood glucose concentration over the preceding 2 weeks. 1.5-Anhydroglucitol in blood is measured as a means of diagnosis and control evaluation of diabetes. Microalbuminuria is widely measured for early detection of diabetic nephropathy. Recently other microproteinuria such as urinary transferrin and IgG are assayed for the same purpose. There are highly significant correlations between microalbuminuria and urinary transferrin or IgG.


General health related quality of life and associated factors among prostate cancer patients in two tertiary medical centers in Kuala Lumpur, Malaysia: a cross-sectional study.
Measurement of quality of life among prostate cancer patients helps the health care providers to understand the impact of the disease in the patients' own perspective. The main aim of this study is to measure the quality of life among prostate cancer patients at University Malaya Medical Center (UMMC) and Universiti Kebangsaan Malaysia Medical Centre (UKMMC) and to ascertain the association factors for physical coefficient summary (PCS) and mental coefficient summary (MCS). A hospital based, cross sectional study using the Short Form-36 (SF-36) questionnaire was conducted over a period of 6 months. A total of 193 respondents were recruited. Their total quality of life score was 70.1± 14.7 and the PCS score was lower compared to MCS. The factors associated for PCS were: age, living partner, renal problem, urinary problem of intermittency, dysuria and hematuria. Factors associated for MCS were: age, living partner, renal problem, presenting prostatic specific antigen and urinary problem of intermittency and dysuria. Our prostate cancer patients had moderate quality of life in the physical health components but their mental health was less affected.


Role of Mad2 expression during the early development of the sea urchin.
Mitotic arrest deficient 2 (Mad2) belongs to the spindle assembly checkpoint (SAC), a mechanism that blocks progression of the cell cycle until microtubule attachment to kinetochores is complete. It has been found to be involved in the resistance of cancer cells to "anti-mitotic" drugs such as paclitaxel. Mad2 controls meiotic progression, but its role during sea urchin development had never been investigated. Furthermore, the existence of a SAC in this species had never been proved. The present data show that a Mad2 protein, highly homologous to that of humans, is expressed in this species. Mad2 expression increases during development, becoming confined to the endomesoderm at gastrula stages. The level of Mad2 expression is enhanced in embryos that do not gastrulate after treatment with anti-mitotic drugs, lithium or inhibition of the ERK pathway. Mis-aligned and lagging chromosomes were induced after injection of an anti-Mad2 antibody or a Mad2 morpholino. Our results point to the role of a non-canonical SAC involving Mad2 in the control of mitotic divisions of the sea urchin embryo.


Mechanistic studies of cAMP-dependent protein kinase action.
The details of the process by which protein kinase catalyzes phosphoryl group transfers are beginning to be understood. Early work that explored the primary specificity of cAMP-dependent protein kinase action enabled the synthesis of small peptide substrates for the enzyme. Enzyme-peptide interactions seem simpler to understand than protein-protein interactions, so peptide substrates have been used in most protein kinase studies. In most investigations the kinetics for the phosphorylation of small peptides have been interpreted as being consistent with mechanisms which do not invoke phospho-enzyme intermediates (see, for example, Bolen et al.). Protein kinase has been shown to bind two metal ions in the presence of a nucleotide. Using magnetic resonance techniques the binding of these ions has been utilized to elucidate the conformation of nucleotide and peptide substrates or inhibitors when bound in the enzymic active site. Also, two new peptides with the form Leu-Arg-Arg-Ala-Ser-Y-Gly, where Y was either Pro or (N-methyl)Leu, were synthesized and found not to be substrates, within the limits of detection, for protein kinase. The striking lack of affinity that protein kinase has for such peptides which are unlikely to form a beta 3-6 turn has not been reported before. Our results may indicate that this type of turn is a requirement for protein kinase catalyzed phosphorylation or that these peptides lack the ability to form a particular hydrogen bond with the enzyme. Magnetic resonance techniques have indicated that the distance between the phosphorous in the gamma-phosphoryl group of MgATP and the hydroxyl oxygen of serine in the peptide Leu-Arg-Arg-Ala-Ser-Leu-Gly is 5.3 +/- 0.7 A. This, together with certain kinetic evidence, suggests that the mechanism by which protein kinase catalyzes phosphoryl group transfer has considerable dissociative character. Chemical modifications, including one using a peptide-based affinity label, have identified two residues at or near the active site, lysine-72 and cysteine 199. While neither of these groups has been shown to be catalytically essential, similar studies may help to identify groups that are directly involved in the catalytic process. Finally, a spectrophotometric assay for cAMP-dependent protein kinase has been described. Using this assay the preliminary results of an in-depth study of the pH dependence of protein kinase catalyzed phosphoryl group transfer have been obtained. This study shall aid in the identification of active site residues and should contribute to the elucidation of the enzyme's catalytic mechanism.(ABSTRACT TRUNCATED AT 400 WORDS)


Ex Vivo Imaging of Resident CD8 T Lymphocytes in Human Lung Tumor Slices Using Confocal Microscopy.
CD8 T cell are key players in the fight against cancer. In order for CD8 T cells to kill tumor cells they need to enter into the tumor, migrate within the tumor microenvironment and respond adequately to tumor antigens. The recent development of improved imaging approaches, such as 2-photon microscopy, and the use of powerful mouse tumor models have shed light on some of the mechanisms that regulate anti-tumor T cell activities. Whereas such systems have provided valuable insights, they do not always predict human responses. In human, our knowledge in the field mainly comes from a description of fixed tumor samples from human patients, as well as in vitro studies. However, in vitro models lack the complex three-dimensional tumor milieu and, therefore, are incomplete approximations of in vivo T cell activities. Fresh slices made from explanted tissue represent a complex multi-cellular tumor environment that can act as an important link between co-cultured studies and animal models. Originally set up in murine lymph nodes1 and previously described in a JoVE article2, this approach has now been transposed to human tumors to examine the dynamics of both plated3 as well as resident T cells4. Here, a protocol for the preparation of human lung tumor slices, immunostaining of resident CD8 T and tumor cells, and tracking of CD8 T lymphocytes within the tumor microenvironment using confocal microscopy is described. This system is uniquely placed to screen for novel immunotherapy agents favoring T cell migration in tumors.


Elucidation of Physiological, Transcriptomic and Metabolomic Salinity Response Mechanisms in Medicago sativa.
Alfalfa (Medicago sativa L.) is a widely grown perennial leguminous forage crop with a number of positive attributes. However, despite its moderate ability to tolerate saline soils, which are increasing in prevalence worldwide, it suffers considerable yield declines under these growth conditions. While a general framework of the cascade of events involved in plant salinity response has been unraveled in recent years, many gaps remain in our understanding of the precise molecular mechanisms involved in this process, particularly in non-model yet economically important species such as alfalfa. Therefore, as a means of further elucidating salinity response mechanisms in this species, we carried out in-depth physiological assessments of M. sativa cv. Beaver, as well as transcriptomic and untargeted metabolomic evaluations of leaf tissues, following extended exposure to salinity (grown for 3-4 weeks under saline treatment) and control conditions. In addition to the substantial growth and photosynthetic reductions observed under salinity treatment, we identified 1233 significant differentially expressed genes between growth conditions, as well as 60 annotated differentially accumulated metabolites. Taken together, our results suggest that changes to cell membranes and walls, cuticular and/or epicuticular waxes, osmoprotectant levels, antioxidant-related metabolic pathways, and the expression of genes encoding ion transporters, protective proteins, and transcription factors are likely involved in alfalfa's salinity response process. Although some of these alterations may contribute to alfalfa's modest salinity resilience, it is feasible that several may be disadvantageous in this context and could therefore provide valuable targets for the further improvement of tolerance to this stress in the future.


Benign pleomorphic adenomas of the salivary glands: surface coil MR imaging versus CT.
High resolution magnetic resonance (MR) imaging of three salivary gland tumors was performed at 1.5 T and compared with CT. Two of the three tumors were well seen on CT. However, one required CT sialography to separate tumor from the remaining parotid parenchyma, and, in the other, soft tissue invasion could not be excluded on the basis of CT. Magnetic resonance imaging demonstrated sharp tumor margins with no evidence of invasion. All three tumors were pathologically proven benign pleomorphic adenomas confirming the MR findings. On T2 weighted images, the two large adenomas demonstrated inhomogeneity that was not observed on CT.


Transient immune suppression of inapparent carriers infected with a principal neutralizing domain-deficient equine infectious anaemia virus induces neutralizing antibodies and lowers steady-state virus replication.
The genetic variation of equine infectious anaemia virus (EIAV) clearly affects the antigenic properties of the viral envelope; however, effects on immunogenicity remain undefined, although widely assumed. Here, the immunogenicity is reported of a novel, neutralization-resistant, pony-isolate envelope EIAV(PV564DeltaPND) that contains a 14-residue deletion in the designated principal neutralizing domain (PND) of the gp90 protein. Two ponies inoculated with a chimeric virus, EIAV(DeltaPND), containing the EIAV(PV564DeltaPND) envelope in a reference provirus strain, remained asymptomatic through 14 months post-inoculation, producing high steady-state levels of envelope-specific antibodies but no detectable serum-neutralizing antibodies. Consequent dexamethasone-induced immune suppression produced characteristic EIA that resolved concomitantly with the development of high-titre, strain-specific, neutralizing antibodies and a 100-fold reduction in steady-state virus loads. These results demonstrate: natural variations in the EIAV envelope have profound effects on both antigenic and immunogenic properties; the PND is not required for neutralizing antibody responses; and transient immune suppression can enhance established host immunity to achieve more effective control of steady-state lentivirus replication.


Perinatal outcome associated with nuchal umbilical cord.
Nuchal umbilical cord (NUC) is a possible complication of pregnancy which can be associated with adverse perinatal outcome. A retrospective study was done at the County Emergency Hospital Timisoara, Romania, between January 2009 and December 2010 and included cases with NUC at the time of delivery. Outcome variable related to the mothers and newborns were studied. The incidence of NUC in the studied period was 8%. Most were primiparous. There were no significant statistical differences between vaginal births and cesarean section with one minute Apgar scores higher than 8. Five percent of the studied group presented intrauterine fetal death. The presence of NUC implies more attention but are not associated with increased rate of operative vaginal or cesarean delivery. One minute Apgar scores in these cases are comparable. NUC can be a cause of IUFD.


Dual-Phase Separation in a Semiconfined System: Monodispersed Heterogeneous Block-Copolymer Membranes for Cell Encoding and Patterning.
Block copolymers (BCPs) have the capacity to self-assemble into a myriad of well-defined aggregate structures, offering great promise for the construction of drug delivery, photolithographic templates, and complex nanoscale assemblies. A uniqueness of these materials is their propensity to become kinetically frozen in non-equilibrium states, implying that the process of self-assembly can be utilized to remodel the resulting structures. Here, a new semiconfined system for processing the BCP self-assembly is constructed, in which an unusual dual-phase separation occurs, including nonsolvent-induced microphase separation and osmotically driven macrophase separation, ultimately yielding heterogeneous BCP membranes. These membranes with cellular dimensions show unique anisotropy that can be used for cell encoding and patterning, which are highly relevant to biology and medicine. This processing method not only provides new levels of tailorability to the structures and encapsulated contents of BCP assemblies, but can also be generalized to other block polymers, particularly those with attractive electronic and/or optical properties.


Local vs global approaches to treat two equivalent methyl internal rotations and 14N nuclear quadrupole coupling of 2,5-dimethylpyrrole.
The microwave spectrum of 2,5-dimethylpyrrole was recorded using a molecular jet Fourier transform microwave spectrometer operating in the frequency range from 2 to 26.5 GHz. Only one stable conformer was observed as expected and confirmed by quantum chemical calculations carried out to complement the experimental analysis. The two equivalent methyl groups cause each rotational transition to split into four torsional species, which is combined with the quadrupole hyperfine splittings in the same order of magnitude arising from the 14N nucleus. This results in a complicated spectrum feature. The spectral assignment was done separately for each torsional species. Two global fits were carried out using the XIAM code and the BELGI-C2v-2Tops-hyperfine code, a modified version of the BELGI-C2v-2Tops code, giving satisfactory root-mean-square deviations. The potential barriers to internal rotation of the two methyl groups were determined to be V3 = 317.208(16) cm-1. The molecular parameters were obtained with high accuracy, providing all necessary ground state information for further investigations in higher frequency ranges and on excited torsional-vibrational states.


[The influence of percutaneous nephrolithotomy on renal function evaluated by 99m-Tc-DTPA renoscintigram].
The influence of percutaneous nephrolithotomy (PNL) on renal function was evaluated by 99m-Tc-DTPA renoscintigraphy. The renoscintigram was recorded and regions of interest of both kidneys (R) and background (BKG) were settled. Renal uptake ratio was calculated by the following formula. Renal uptake ratio = (formula; see text). The renal uptake ratio of 50 patients were compared before and up to 6 months after PNL. Most cases showed an increase or no change in post-PNL renal uptake ratio in operated kidney. Five cases (10%) showed decreased renal uptake ratio of post-PNL. No risk factors for the renal damage caused by PNL such as age, sex, stone size, number of nephrostomy and course of PNL were detected when compared with increased or no change groups. In cases of complete staghorn calculi in the decreased renal uptake ratio group, compensatory hypertrophy of contralateral kidney PNL course might occur, and the function of the operated kidney might not have recovered. More during cases and longer follow-up is necessary for the recovery of renal function.