[The peculiarities of the chromosome organization in meiosis].
Meiotic and mitotic chromosomes differ in a number of features. 1. At the early prophase I of meiosis, chromosomes acquire proteinaceous axial elements (AEs) which were absent at mitosis. In addition to somatic cohesins, AEs contain meiosis-specific cohesins REC8, SMC1beta, STAG3. 2. At the middle prophase I, proteinaceous lateral elements (LEs) of synaptonemal complexes (SC) are shaped on a basis of AEs. Proteins of LEs are not conserved, but in Saccharomyces cerevisiae and Arabidopsis thaliana they contain functional domains with conserved secondary structure. Proteins or functional domains similar to SC proteins have been found in green and brown algae, some of lower fungi and in Coelenterata amongs about 679 hundreds of proteins of primitive eukaryotes studied with bioinformatic methods. 3. During the pachytene and diplotene stages of meiosis, chromosomes of spermatocytes and mother pollen cells acquire the structure resembling in miniature the structure of amphibian and avian lamp brush chromosomes. Lateral chromatin loops of 90, 160 and more than 480 Kb in size are observed in human spermatocytes during the diplotene stage. Taken together, these findings support the idea of considerable conservation of the scheme of molecular and ultrastructural organization of meiotic chromosomes in a variety of eukaryotic organisms.


Sympathetic-Sensory Coupling as a Potential Mechanism for Acupoints Sensitization.
A series of studies have demonstrated acupoint sensitization, in which acupoints can be activated in combination with sensory hypersensitivity and functional plasticity during visceral disorders. However, the mechanisms of acupoint sensitization remain unclear. Neuroanatomy evidence showed nociceptors innervated in acupoints contribute to the mechanism of acupoint sensitization. Increasing studies suggested sympathetic nerve plays a key role in modulating sensory transmission by sprouting or coupling with sensory neuron/nociceptor in the peripheral, forming the functional structure of the sympathetic-sensory coupling. Notably, the sensory inputs of the disease-induced sensitized acupoint contribute to the homeostatic regulation and also involve in delivering therapeutic information under acupuncture, hence, the role of sprouted sympathetic in acupoint function should be given attention. We herein reviewed the current knowledge of sympathetic and its sprouting in pain modulation, then discussed and highlighted the potential value of sympathetic-sensory coupling in acupoint functional plasticity.


Prospective exploratory study of gemcitabine and S-1 against elderly patients with advanced non-small cell lung cancer.
A prospective study was conducted to investigate the efficacy of a combined regimen of gemcitabine and S-1 for the treatment of elderly patients (>70 years) with advanced non-small cell lung cancer (NSCLC) as a first-line setting based on the dosage recommended in a previous phase I study. Chemotherapy-naïve patients with advanced NSCLC received gemcitabine plus S-1. S-1 (40 mg/m2) was administered orally twice daily for 14 days while gemcitabine (1,000 mg/m2) was administered on days 1 and 15 of each cycle, and this regimen was repeated every 4 weeks. A total of 20 patients were included in the present study. Of these, 8 patients achieved an overall response rate of 40.0%, and the overall disease control rate was 65.0%. According to the histological type, the response rate in patients with NSCLC and adenocarcinoma was 38.5%, and that for non-adenocarcinoma was 42.9%. Progression-free survival and median survival times were 6.4 months and 17.8 months, respectively. Grade 3 or 4 hematological toxicities observed were leukopenia (29%) and neutropenia (24%), while febrile neutropenia was not observed in any patient. The only non-hematological adverse event observed was grade 3 skin rash (10%). Therefore, the combination of gemcitabine and S-1 may be a promising and feasible regimen in the first-line setting for elderly patients with advanced NSCLC.


PLD1 participates in BDNF-induced signalling in cortical neurons.
The brain-derived neurotrophic factor BDNF plays a critical role in neuronal development and the induction of L-LTP at glutamatergic synapses in several brain regions. However, the cellular and molecular mechanisms underlying these BDNF effects have not been firmly established. Using in vitro cultures of cortical neurons from knockout mice for Pld1 and Rsk2, BDNF was observed to induce a rapid RSK2-dependent activation of PLD and to stimulate BDNF ERK1/2-CREB and mTor-S6K signalling pathways, but these effects were greatly reduced in Pld1(-/-) neurons. Furthermore, phospho-CREB did not accumulate in the nucleus, whereas overexpression of PLD1 amplified the BDNF-dependent nuclear recruitment of phospho-ERK1/2 and phospho-CREB. This BDNF retrograde signalling was prevented in cells silenced for the scaffolding protein PEA15, a protein which complexes with PLD1, ERK1/2, and RSK2 after BDNF treatment. Finally PLD1, ERK1/2, and RSK2 partially colocalized on endosomal structures, suggesting that these proteins are part of the molecular module responsible for BDNF signalling in cortical neurons.


Posterior laryngeal cleft.
The posterior laryngeal cleft is a rare congenital anomaly which results from absence of fusion of the posterior cricoid lamina and possibly the tracheoesophageal septum. A high degree of suspicion is required for early diagnosis and repair to prevent significant morbidity and mortality from recurrent aspiration pneumonia. Minor clefts may be repaired endoscopically, whereas severe laryngotracheoesophageal clefts require a multidisciplinary approach with combined extrathoracic and intrathoracic repair. Associated congenital anomalies increase the mortality of the laryngeal clefts and may complicate the timing of surgical repair.


Recent Progress in Small-Molecule Fluorescence and Photoacoustic Dual-Modal Probes for the In-Vivo Detection of Bioactive Molecules.
Intracellular bioactive molecules are essential for the maintenance of homeostasis in living organisms. Abnormal levels of them are closely related to the occurrence and development of some diseases. Hence, the direct and accurate visualization of these bioactive molecules is of vital importance for exploring their pathological roles. However, the low-content, short-lived, and widely distributed properties of bioactive molecules impede the comprehensive analysis of them dramatically. Fluorescence and photoacoustic dual-mode imaging technology provides a new solution to the above issue. Specifically, the combination of fluorescence and photoacoustic, which possesses the advantages of high resolution and in-depth tissue analysis, enables a more in-depth and systematic exploration of the pathogenic mechanisms of bioactive molecules. Moreover, due to the structural tailorability of small molecule probes, numerous small molecule dual-mode probes have been developed to meet the demand for real-time tracking and visualization of bioactive molecules in living cells or in vivo. Hence, in this review, we briefly summarize the key advances in small molecule fluorescence and photoacoustic dual-modal probes within recent years (2015-2021). A particular focus is placed on the design strategies and biological applications of probes for the detection of various bioactive molecules in vivo. Furthermore, the challenges and further prospects in this hot field are highlighted.


Catecholaminergic influences on motion sickness.
The evidence for possible roles of catecholamines in motion sickness is reviewed. l-Dopa is implicated in that it is the precursor to catecholaminergic neurotransmitters. Dopamine may have a role in stimulating motion sickness, though much basic information is lacking. Epinephrine may have a stimulatory role, since anatomical, pharmacological and physiological evidence is consistent with such a role. Norepinephrine probably inhibits motion sickness, as indicated by anatomical, physiological and pharmacological evidence. Information is missing that is critical to the understanding of the roles of each neurotransmitter in motion sickness.


Vaspin as a Risk Factor of Insulin Resistance in Obstructive Sleep Apnea-Hypopnea Syndrome in an Animal Model.
In this study, we aimed to establish a chronic intermittent hypoxia model in rats and explore the possible role of vaspin in insulin sensitivity. Healthy male Wistar rats were randomly divided into two groups: normal control group (NC) and chronic intermittent hypoxia group (CIH). The NC group was raised under physiological conditions and the CIH group was kept in the plexiglass chamber between 9 am and 5 pm undergoing intermittent hypoxic challenge for 8 hours/day for 8 weeks. Arterial blood pressure of rats (tail cannulation) was measured before and after the study. Fasting plasma glucose (FPG), total cholesterol (TC), triglycerides (TG), fasting insulin (FINS), vaspin, and leptin levels were measured. Vaspin mRNA expression in visceral adipose tissues was measured with Real Time-PCR. The protein levels of vaspin, Akt and phospho-Akt in visceral tissues were determined by Western-blot. At baseline, all the measurements in the CIH and NC groups were comparable. By the end of the experiment, the blood pressure of the CIH group was significantly higher than the NC group. The levels of FPG, FINS, TG, TC, leptin, and vaspin in the CIH group were significantly higher than in NC group. Plasma vaspin levels were correlated with FINS, HOMA-IR, and TG levels. Vaspin expression in both mRNA and protein levels in visceral adipose tissues of the CIH group were clearly higher than the NC group. Phospho-Akt protein level was decreased in visceral adipose tissues of the CIH group compared to the NC group. In the chronic intermittent hypoxia rat model, the expression of vaspin in visceral adipose tissues and plasma were increased, which were correlated with insulin resistance.


Rescue of Recombinant Adenoviruses by CRISPR/Cas-Mediated in vivo Terminal Resolution.
Recombinant adenovirus (rAd) vectors represent one of the most frequently used vehicles for gene transfer applications in vitro and in vivo. rAd genomes are constructed in Escherichia coli where their genomes can be maintained, propagated, and modified in form of circular plasmids or bacterial artificial chromosomes. Although the rescue of rAds from their circular plasmid or bacmid forms is well established, it works with relatively low primary efficiency, preventing this technology for library applications. To overcome this barrier, we tested a novel strategy for the reconstitution of rAds that utilizes the CRISPR/Cas-machinery to cleave the circular rAd genomes in close proximity to their inverted terminal repeats (ITRs) within the producer cells upon transfection. This CRISPR/Cas-mediated in vivo terminal resolution allowed efficient rescue of vectors derived from different human adenovirus (HAdV) species. By this means, it was not only possible to increase the efficiency of virus rescue by about 50-fold, but the presented methodology appeared also remarkably simpler and faster than traditional rAd reconstitution methods.


Intramolecular electron transfer in yeast flavocytochrome b2 upon one-electron photooxidation of the fully reduced enzyme: evidence for redox state control of heme-flavin communication.
Flavocytochrome b2, which has been fully reduced using L-lactate, can be rapidly oxidized by 1 equiv using the laser-generated triplet state of 5-deazariboflavin. Parallel photoinduced oxidation occurs at the reduced heme and at the fully reduced FMN (FMNH2) prosthetic groups of different enzyme monomers, producing the anion semiquinone of FMN and a ferric heme. Following the initial oxidation reaction, rapid intramolecular reduction of the ferric heme occurs with concomitant oxidation of FMNH2, generating the neutral FMN semiquinone. The observed rate constant for this intramolecular electron transfer is 2200 s-1, which is 1 order of magnitude larger than the turnover number under these conditions. A slower reduction of the heme prosthetic group also occurs with an observed rate constant of approximately 10 s-1, perhaps due to intersubunit electron transfer from reduced FMN to heme. The rapid intramolecular electron transfer between the FMNH2 and ferric heme is eliminated upon addition of excess pyruvate (Ki = 3.8 mM). This latter result indicates that pyruvate inhibition of catalytic turnover apparently can occur at the FMNH2-->heme electron transfer step. These results markedly differ from those previously obtained (Walker, M. C., & Tollin, G. (1991) Biochemistry 30, 5546-5555) and confirmed here for electron transfer within the one-electron reduced enzyme and for the effect of pyruvate binding, suggesting that intramolecular communication between the heme and flavin prosthetic groups can be controlled by the redox state of the enzyme and by ligand binding to the active site.


NF-kappaB and STAT3 signaling in glioma: targets for future therapies.
Glioblastoma remains the most clinically challenging tumor of the CNS, as evidenced by the dismal change in overall survival over the past 50 years. However, recent advances in high-throughput screening techniques have given rise to a wealth of new information regarding the aberrant signaling pathways that drive the tumor phenotype. Two of these so-called 'oncopathways' are NF-kappaB and JAK/STAT. This review will describe the basic mechanisms of these pathways, explore the relevance of NF-kappaB and JAK/STAT signaling in glioblastoma, and look ahead to experimental compounds that will integrate our knowledge of these pathways into existing therapies.


Markov Property of the Conformal Field Theory Vacuum and the a Theorem.
We use strong subadditivity of entanglement entropy, Lorentz invariance, and the Markov property of the vacuum state of a conformal field theory to give new proof of the irreversibility of the renormalization group in d=4 space-time dimensions-the a theorem. This extends the proofs of the c and F theorems in dimensions d=2 and d=3 based on vacuum entanglement entropy, and gives a unified picture of all known irreversibility theorems in relativistic quantum field theory.


Exploring the relationship between head anatomy and cochlear implant stability in children.
In our experience, surgical outcomes in children have been excellent with a low complication rate. Our aim in this study was to better understand what aspects of our current surgical technique have been successful with a view to retain those that are beneficial as we proceed with implantation of future devices. Because the receiver-stimulator and overlying skin flap may be more vulnerable to damage in children than adults, we concentrated on issues related to the positioning and security of this part of the implant on the head. Three specific areas of vulnerability were explored in separate experiments. In Experiment 1, we determined the effect of the position of the device on the ability of a child to roll their head without allowing contact between the device and a supporting surface. The 'freeroll' angle was determined for devices position conventionally (back position) and for those in which the device is placed in a more anterior position (up position). In Experiment 2, we studied the retentive capacity of the child's pericranium and measured the displacement force required to dislodge an implant from the bed if retained by the calvarium only. In Experiment 3, we compared the skull curvature of children in whom the device was placed in the back versus the up position. These results inform us as how to best proceed with implantation in children using future devices that have thinner and wider receiver-stimulators.


Study of the influence of coating methods on lipid spheres manufactured on rotor fluidized bed process.
Different previous works have shown that various kinds of spheres can be manufactured by rotor granulation in a 'single-pot process' using a lipid base: hydrogenated castor oil. This single-pot technology is based on wet granulation where all components are placed in the powder form in the rotor bowl; then, they are continuously suspended in a fluidized air, with a tangentially sprayed liquid solution. This process allows the granulation and manufacturing of sphere during the same time. Previous experiments have studied the influence of the formulation and the manufacturing process parameters on spheres in terms of feasibility and dissolution properties. Both the spraying time and the weight of liquid sprayed were found to be the most relevant parameters that govern the final quality of the sphere. Now, in a second part of the work, a first comparison is made with two different fluid bed methods: the tangential rotor spray and the Wurster bottom spray for coating the lipid spheres previously manufactured with the rotor tangential spray. The external aspect of the coated spheres manufactured has been evaluated with an electronic microscopy analysis and a study of dissolution properties of the active ingredient has been done by USP in vitro dissolution tests.


Insight into the mechanisms and functions of spliceosomal snRNA pseudouridylation.
Pseudouridines (Ψs) are the most abundant and highly conserved modified nucleotides found in various stable RNAs of all organisms. Most Ψs are clustered in regions that are functionally important for pre-mRNA splicing. Ψ has an extra hydrogen bond donor that endows RNA molecules with distinct properties that contribute significantly to RNA-mediated cellular processes. Experimental data indicate that spliceosomal snRNA pseudouridylation can be catalyzed by both RNA-dependent and RNA-independent mechanisms. Recent work has also demonstrated that pseudouridylation can be induced at novel positions under stress conditions, suggesting a regulatory role for Ψ.


High-energy x-ray diffractometer for nondestructive strain depth profile measurement.
We describe a lab-based high-energy x-ray diffraction system and a new approach to nondestructively measuring strain profiles in polycrystalline samples. This technique utilizes the tungsten K(α1) characteristic radiation from a standard industrial x-ray tube. We introduce a simulation model that is used to determine strain values from data collected with this system. Examples of depth profiling are shown for shot peened aluminum and titanium samples. Profiles to 1 mm depth in aluminum and 300 μm depth in titanium with a depth resolution of 20 μm are presented.


Video and audio processing in paediatrics: a review.
Video and sound acquisition and processing technologies have seen great improvements in recent decades, with many applications in the biomedical area. The aim of this paper is to review the overall state of the art of advances within these topics in paediatrics and to evaluate their potential application for monitoring in the neonatal intensive care unit (NICU). For this purpose, more than 150 papers dealing with video and audio processing were reviewed. For both topics, clinical applications are described according to the considered cohorts-full-term newborns, infants and toddlers or preterm newborns. Then, processing methods are presented, in terms of data acquisition, feature extraction and characterization. The paper first focuses on the exploitation of video recordings; these began to be automatically processed in the 2000s and we show that they have mainly been used to characterize infant motion. Other applications, including respiration and heart rate estimation and facial analysis, are also presented. Audio processing is then reviewed, with a focus on the analysis of crying. The first studies in this field focused on induced-pain cries and the newest ones deal with spontaneous cries; the analyses are mainly based on frequency features. Then, some papers dealing with non-cry signals are also discussed. Finally, we show that even if recent improvements in digital video and signal processing allow for increased automation of processing, the context of the NICU makes a fully automated analysis of long recordings problematic. A few proposals for overcoming some of the limitations are given.


Comparison of the neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios in normoglycemic and hyperglycemic subjects.
To compare the neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios as possible parameters of systemic inflammation in hyperglycemic and normoglycemic subjects. A retrospective, cross-sectional study of data collected from patients tested for fasting blood glucose, glycated hemoglobin (HbA1c) and blood count on the same day, between July and December 2016. Patients were divided into hyperglycemic and normoglycemic, and matched by age and sex. The data were analyzed using Epi Info™, version 184.108.40.206, for the Windows® platform. We enrolled 278 subjects, 139 hyperglycemic and 139 normoglycemic. The absolute number of leukocytes and neutrophils was higher in the Hyperglycemic Group (p=0.006 and p=0.004, respectively). There was no difference in the neutrophil-to-lymphocyte ratio between the Hyperglycemic Group and the Normoglycemic Group (2.1 versus 2.0; p=0.264), and both neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios showed no differences between those with HbA1c ≥7% (n=127, p=0.778) and those with HbA1c <7% (n=12, p=0.490). In contrast, the platelet-to-lymphocyte ratio was lower in the Hyperglycemic Group (117.8 versus 129.6; p=0.007). Hyperglycemic subjects had a neutrophil-to-lymphocyte ratio similar to that of normoglycemic subjects, but had a lower platelet-to-lymphocyte ratio. Future prospective studies will be useful to determine the importance and prognostic value of neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios in the hyperglycemic state.


Functional differences between the Oct2 transactivation domains determine the transactivation potential of individual Oct2 isoforms.
The lymphocyte specific transcription factor Oct2 is involved in mediating the B-cell specific transcriptional activity of the octamer motif. Mutational analyses in the context of the complete Oct2 protein had indicated that Oct2 contains two transactivation domains. These two domains appeared to be redundant for activation from a promoter proximal position, whereas stimulation from a remote enhancer position specifically required the C-terminal transactivation domain and an additional B-cell restricted activity. We have generated fusion proteins between the DNA binding domain of the yeast Gal4 transcription factor and individual Oct2 protein domains to analyze their transactivation potential separately. We show that both N- and C-terminal domains can stimulate transcription from a promoter proximal position independently. However, only the C-terminal transactivation domain activates from a distance and it can only do so in B-cells. The C-terminal transactivation domain represents a composite transactivation domain. Whereas removal of just 9 aminoacids from the extreme C-terminus lead to a complete inactivation of this domain deletions from the other side resulted in a gradual loss of activity. We also characterized the transactivation potential of different N-terminal regions of Oct2 generated by alternative splicing. We show that the N-terminus of one of the isoforms, Oct2.3, contains a negative regulatory domain (NRD), which can inactivate the neighbouring glutamine-rich transactivation in cis. The presence of this NRD affects the overall phosphorylation state of the Oct2 protein. This result suggests that the mechanism of inactivation might involve differential protein phosphorylation.


Looking beyond appearances: when liver biopsy is the key for hepatic tuberculosis diagnosis.
Primary hepatic tuberculosis is a rare clinical entity with non-specific clinical and imaging features that can mimic other liver diseases, representing a diagnostic challenge. We report a case of a 35-year-old man with metabolic syndrome, type 2 diabetes and high alcohol consumption presenting asymptomatic with abnormal liver tests, hepatosplenomegaly and diffuse hepatic steatosis in ultrasound imaging initially suspected to be alcoholic steatohepatitis but later diagnosed as hepatic tuberculosis in the histological specimen. Anti-tuberculosis therapy was started. This clinical case highlights the diagnostic difficulty of hepatic tuberculosis and the importance of not overlooking liver biopsy and to consider it in the differential diagnosis in patients with obvious hepatic injury factors but with atypical clinical presentation.


Developmental defects of the thyroid gland: relationship with advanced maternal age.
Developmental defects of the thyroid gland are the most frequent causes of permanent congenital hypothyroidism. This study aimed to investigate the epidemiological features of patients with thyroid dysgenesis (TD). Medical records of 234 patients with TD followed between the years 2008 and 2010 were evaluated retrospectively. Diagnosis was made by ultrasonography. Of 234 patients, 120 (51.3%) were male and 114 (48.7%) were female. Male to female ratio was 1.08 and there were no significant differences in epidemiologic and clinical findings between girls and boys. One hundred eighty-three patients (78.2%) were diagnosed as hypoplasia, 35 (14.9%) as thyroid agenesis, 4 as ectopic thyroid gland and 12 as hemiagenesis. The mean maternal age of the group was 28.9 ± 0.4 years (range 18 to 45 years), which is significantly higher than the recently reported mean maternal ages for Turkish women. Advanced maternal age was more prevalent in patients with TD. Our clinical and epidemiologic findings suggested no evidence of sexual dimorphism.


Molecular characterization of the NSP4 gene of human group A rotavirus samples from the West Central region of Brazil.
Nonstructural protein 4 (NSP4), encoded by group A rotavirus genome segment 10, is a multifunctional protein and the first recognized virus-encoded enterotoxin. The NSP4 gene has been sequenced, and five distinct genetic groups have been described: genotypes A-E. NSP4 genotypes A, B, and C have been detected in humans. In this study, the NSP4-encoding gene of human rotavirus strains of different G and P genotypes collected from children between 1987 and 2003 in three cities of West Central region of Brazil was characterized. NSP4 gene of 153 rotavirus-positive fecal samples was amplified by reverse transcriptase-polymerase chain reaction and then sequenced. For phylogenetic analysis, NSP4 nucleotide sequences of these samples were compared to nucleotide sequences of reference strains available in GenBank. Two distinct NSP4 genotypes could be identified: 141 (92.2%) sequences clustered with NSP4 genotype B, and 12 sequences (7.8%) clustered with NSP4 genotype A. These results reinforce that further investigations are needed to assess the validity of NSP4 as a suitable target for epidemiologic surveillance of rotavirus infections and vaccine development.


Pharmacological interventions for recurrent abdominal pain in childhood.
Between 4% and 25% of school-aged children at some stage complain of recurrent abdominal pain (RAP) of sufficient severity to interfere with their daily lives. When no clear organic cause is found, the children are managed with reassurance and simple measures; a large range of pharmacological interventions have been recommended for use in these children. To determine the effectiveness of pharmacological interventions for RAP in children of school age. We searched the Cochrane Central Register of Controlled Trials (CENTRAL), Ovid MEDLINE, Embase, and eight other electronic databases up to June 2016. We also searched two trials registers and contacted researchers of published studies. Randomised controlled trials involving children aged five to 18 years old with RAP or an abdominal pain-related functional gastrointestinal disorder, as defined by the Rome III criteria (Rasquin 2006). The interventions were any pharmacological intervention compared to placebo, no treatment, waiting list, or standard care. The primary outcome measures were pain intensity, pain duration or pain frequency, and improvement in pain. The secondary outcome measures were school performance, social or psychological functioning, and quality of daily life. Two review authors independently screened titles, abstracts, and potentially relevant full-text reports for eligible studies. Two review authors extracted data and performed a 'Risk of bias' assessment. We used the GRADE approach to rate the overall quality of the evidence. We deemed a meta-analysis to be not appropriate as the studies were significantly heterogeneous. We have consequently provided a narrative summary of the results. This review included 16 studies with a total of 1024 participants aged between five and 18 years, all of whom were recruited from paediatric outpatient clinics. Studies were conducted in seven countries: seven in the USA, four in Iran, and one each in the UK, Switzerland, Turkey, Sri Lanka, and India. Follow-up ranged from two weeks to four months. The studies examined the following interventions to treat RAP: tricyclic antidepressants, antibiotics, 5-HT4 receptor agonists, antispasmodics, antihistamines, H2 receptor antagonists, serotonin antagonists, selective serotonin re-uptake inhibitors, a dopamine receptor antagonist, and a hormone. Although some single studies reported that treatments were effective, all of these studies were either small or had key methodological weaknesses with a substantial risk of bias. None of these 'positive' results have been reproduced in subsequent studies. We judged the evidence of effectiveness to be of low quality. No adverse effects were reported in these studies. There is currently no convincing evidence to support the use of drugs to treat RAP in children. Well-conducted clinical trials are needed to evaluate any possible benefits and risks of pharmacological interventions. In practice, if a clinician chooses to use a drug as a 'therapeutic trial', they and the patient need to be aware that RAP is a fluctuating condition and any 'response' may reflect the natural history of the condition or a placebo effect, rather than drug efficacy.


Objective analysis of intermediate-term outcome of the Ponseti technique: a review of the experience from Los Angeles.
The Ponseti method of manipulative treatment for clubfoot deformity became widely adopted by pediatric orthopaedic surgeons beginning in the mid-1990s. The technique allows correction of most idiopathic clubfeet using gentle manipulation and cast application. The treatment represents a marked advance over past efforts to gain correction of the foot through extensive release surgery. In 2006, we began a Clubfoot Clinic at the Orthopaedic Institute for Children in Los Angeles, California dedicated to managing clubfoot patients using Ponseti's method. An IRB-approved database of patient-related, treatment related, and demographic variables was assembled and used to ascertain the outcome of treatment as well as to address parental questions regarding certain aspects of treatment. Here, we present a review of our body of work, which has improved clinical decision making as well as our ability to better inform our patients' parents regarding the treatment and prognosis of the Ponseti method. Studies from our institution showed that while relapses and the need for extra-articular tibialis anterior tendon transfer (TATT) surgery remain common to the Ponseti method, these events do not adversely affect overall patient function or satisfaction. These findings were not unlike those of classic studies reported from Ponseti's institution. We conclude that the Ponseti method is not only a technique to achieve initial correction of an idiopathic clubfoot, but also how to manage relapses that will inevitably occur in many patients. While relapses and tendon transfer surgery are likely to remain common with this treatment method, these events do not adversely affect overall patient function or satisfaction. The parents of infants whose clubfeet are managed using the Ponseti method should be counselled accordingly.


The effects of intravenous lignocaine on depth of anaesthesia and intraoperative haemodynamics during open radical prostatectomy.
Lignocaine is a local anaesthetic agent, which is also commonly used as a perioperative analgesic adjunct to accelerate rehabilitation and enhance recovery after surgery. Lignocaine's systemic effects on intraoperative haemodynamics and volatile anaesthetic requirements are not well explored. Therefore, we evaluated the effects of intravenous lignocaine on intraoperative volatile agent requirements and haemodynamics in patients undergoing major abdominal surgery. We performed an analysis of 76 participants who underwent elective open radical retropubic prostatectomy. Patients received lignocaine (1.5 mg/kg loading dose) followed by an infusion (1.5 mg/kg/h) for the duration of surgery, or saline at an equivalent rate. The aims of the study were to evaluate the end-tidal sevoflurane concentration required to maintain a bispectral index of between 40 and 60. Measurements included intraoperative blood pressure, heart rate, and the volume of intravenous fluids and dosage of vasoactive medications administered. The average end-tidal sevoflurane concentration was lower in the Lignocaine group compared to saline [1.49% (SD: 0.32) vs. 1.89% (SD: 0.29); 95% CI 0.26-0.5, p < 0.001]. In the Lignocaine group, the average mean arterial pressure was 80.3 mmHg (SD: 4.9) compared to 85.1 mmHg (SD: 5.4) in the Saline group (95% CI 2.4-7.1, p < 0.001). Systolic blood pressure was also lower in the Lignocaine group: 121.7 mmHg (SD: 6.1) vs. 128.0 mmHg (SD: 6.4) in the Saline group; 95% CI 3.5-9.2, p < 0.001, as was the mean heart rate [Lignocaine group: 74.9 beats/min (SD: 1.8) vs. 81.5 beats/min (SD: 1.7) in the Saline group, 95% CI 4.1-9.1, p < 0.001]. Maintenance fluid requirements were higher in the Lignocaine group: 3281.1 mL (SD: 1094.6) vs. 2552.6 mL (SD: 1173.5) in the Saline group, 95% CI 206-1251, p = 0.007. There were no differences in the use of vasoactive drugs. Intravenous lignocaine reduces volatile anaesthetic requirements and lowers blood pressure and heart rate in patients undergoing open radical prostatectomy.


Recombinant interleukin-2-activated intracavitary lymphocytes: phenotypic characteristics and effector function.
A preclinical study of intracavitary lymphocytes (ICL) from malignant effusions of cancer patients is described. The object of this study was to evaluate the antitumor potential of ICL as a baseline for developing adoptive immunotherapy trial for ovarian carcinoma patients. The main parameters studied were functional cytolytic activity of fresh and recombinant interleukin-2 (rIL-2)-activated ICL and their phenotypic characteristics. Spontaneous cytolytic activity of ICL was detected in all samples tested against natural killer (NK)-sensitive targets (K562), while very low activity was shown against NK-resistant targets (Daudi) and fresh tumor cells. Activation in culture with rIL-2 generated cytolytic activity against NK-resistant targets and significantly augmented NK activity. The pattern of antitumor lytic activity of ICL resembles lymphokine-activated killer cell activity and is non-major histocompatibility complex restricted against a variety of tumor targets. Phenotypic characterization of fresh ICL showed the predominance of CD3+ cells with the CD4/CD8 ratio resembling that of peripheral blood lymphocytes. During culture with rIL-2, changes in phenotypic expression of activated ICL were detected: enrichment in NKH1+ cells (up to 65%), of CD8+ (up to 70%), and very late antigen (VLA)-1+ cells (up to 54%), concomitantly with a decrease in CD4+ population. The NKH1, CD8, and VLA-1 expression peaked at 2 weeks in culture and coincided with peak cytolytic activity of cultured ICL. Depletion of CD8+ cells from activated ICL resulted in a decreased proportion of cells expressing the NKH1 and VLA-1 phenotype, whereas depletion CD4+ cells led to enrichment in CD8, NKH1, and VLA-1 antigens. Cytolytic activity was significantly increased in CD4 depleted population against NK-resistant and NK-sensitive targets. In this study we found that rIL-2 activation and culture of ICL generates killer activity against NK-resistant and fresh tumor targets and that enrichment in expression of NKH1, CD8, and VLA-1 antigens is associated with effector function.


Nonclassic functions of human topoisomerase I: genome-wide and pharmacologic analyses.
The biological functions of nuclear topoisomerase I (Top1) have been difficult to study because knocking out TOP1 is lethal in metazoans. To reveal the functions of human Top1, we have generated stable Top1 small interfering RNA (siRNA) cell lines from colon and breast carcinomas (HCT116-siTop1 and MCF-7-siTop1, respectively). In those clones, Top1 is reduced approximately 5-fold and Top2alpha compensates for Top1 deficiency. A prominent feature of the siTop1 cells is genomic instability, with chromosomal aberrations and histone gamma-H2AX foci associated with replication defects. siTop1 cells also show rDNA and nucleolar alterations and increased nuclear volume. Genome-wide transcription profiling revealed 55 genes with consistent changes in siTop1 cells. Among them, asparagine synthetase (ASNS) expression was reduced in siTop1 cells and in cells with transient Top1 down-regulation. Conversely, Top1 complementation increased ASNS, indicating a causal link between Top1 and ASNS expression. Correspondingly, pharmacologic profiling showed L-asparaginase hypersensitivity in the siTop1 cells. Resistance to camptothecin, indenoisoquinoline, aphidicolin, hydroxyurea, and staurosporine and hypersensitivity to etoposide and actinomycin D show that Top1, in addition to being the target of camptothecins, also regulates DNA replication, rDNA stability, and apoptosis. Overall, our studies show the pleiotropic nature of human Top1 activities. In addition to its classic DNA nicking-closing functions, Top1 plays critical nonclassic roles in genomic stability, gene-specific transcription, and response to various anticancer agents. The reported cell lines and approaches described in this article provide new tools to perform detailed functional analyses related to Top1 function.


Effect of aging on follicular function may be relieved by exogenous gonadotropin treatment in a sheep model.
The current study investigated hormonal and ovarian changes during physiological reproductive aging in Sarda ewes. In a first experiment, follicular and corpus luteum dynamics were compared during an induced oestrus cycle in aged (12-14 years) and young adult ewes (4-5 years). Oestrus cycle characteristics did not differ between the two experimental groups. However, follicular function during the follicular phase showed significant alterations in aged ewes, as determined by a lack of dominance effect and by lower mean values of circulating oestradiol (E(2)) and inhibin levels, compared with young adult ewes. In a second experiment, differences in follicle growth, hormonal milieu and oocyte quality in response to exogenous FSH administration were assessed in aged and adult ewes. No differences were recorded in ovarian response to FSH treatment between young adult and aged ewes, as evaluated by ultrasonographic data and circulating concentrations of LH, E(2) and inhibin-A. Although the total number of recovered oocytes was similar in the two age groups, the number of good quality oocytes selected for IVM was significantly lower in aged ewes compared with adult ones. Thereafter, no differences were recorded in cleavage rates, total blastocyst output, embryo developmental kinetic and quality between aged and adult groups. In conclusion, this study demonstrated that reproductive aging in sheep is associated with impaired follicle functionality and an increase in the proportion of oocytes showing morphological abnormalities. However interestingly, oocyte developmental competence in vitro and embryo cryotolerance were not affected by the aging process, when only good quality oocytes were chosen.


An argyrophil III method for the demonstration of elastic fibres and membranes.
An esterification with isopropyl alcohol containing 0.2% periodic acid and 2% acetone (at 56 degrees C for 16 hours) followed by a treatment in a special physical developer, similarly, an acetylation with a 3:2 mixture of pyridine and acetic anhydride (at room temperature for 16 hours) followed by the same development, render the elastic fibres and membranes visible. Both pretreatments (esterification and acetylation) serve to make the catalytic points more active in the elastic elements than in the other components of tissue.


High yield production and purification of two recombinant thermostable phosphotriesterase-like lactonases from Sulfolobus acidocaldarius and Sulfolobus solfataricus useful as bioremediation tools and bioscavengers.
Thermostable phosphotriesterase-like lactonases (PLLs) are able to degrade organophosphates and could be potentially employed as bioremediation tools and bioscavengers. But nowadays their manufacturing in high yields is still an issue that limits their industrial applications. In this work we aimed to set up a high yield production and purification biotechnological process of two recombinant PLLs expressed in E. coli, the wild type SacPox from Sulfolobus acidocaldarius and a triple mutated SsoPox C258L/I261F/W263A, originally from Sulfolobus solfataricus. To follow this aim new induction approaches were investigated to boost the enzyme production, high cell density fermentation strategies were set-up to reach higher and higher enzyme yields up to 22-L scale, a downstream train was studied to meet the requirements of an efficient industrial purification process. Physiological studies in shake flasks demonstrated that the use of galactose as inducer increased the enzyme concentrations up to 4.5 folds, compared to the production obtained by induction with IPTG. Optimising high cell density fed-batch strategies the production and the productivity of both enzymes were further enhanced of 26 folds, up to 2300 U·L- 1 and 47.1 U·L- 1·h- 1 for SacPox and to 8700 U·L- 1 and 180.6 U·L- 1·h- 1 for SsoPox C258L/I261F/W263A, and the fermentation processes resulted scalable from 2.5 to 22.0 L. After being produced and extracted from the cells, the enzymes were first purified by a thermo-precipitation step, whose conditions were optimised by response surface methodology. A following ultra-filtration process on 100 and 5 KDa cut-off membranes drove to a final pureness and a total recovery of both enzymes of 70.0 ± 2.0%, suitable for industrial applications. In this paper, for the first time, a high yield biotechnological manufacturing process of the recombinant enzymes SacPox and SsoPox C258L/I261F/W263A was set-up. The enzyme production was boosted by combining a new galactose induction approach with high cell density fed-batch fermentation strategies. An efficient enzyme purification protocol was designed coupling a thermo-precipitation step with a following membrane-based ultra-filtration process.


Improving the productivity of 19,20-epoxy-cytochalasin Q in Xylaria sp. sof11 with culture condition optimization.
19,20-Epoxy-cytochalasin Q (B5A) is a cytochalasin with a wide range of biological activities, which can be produced by Xylaria sp. sof11, a strain isolated from the seafloor of the northern South China Sea. Since the low titer of B5A has greatly limited its further studies, we have systematically conducted the fermentative optimization for B5A production in this article. The effects of major medium components, including the carbon and organic nitrogen sources, as well as of the concentration of sea salt, were respectively investigated through single-factor experiments. As a result, sucrose and fish meal were determined to be the key factors affecting the production of B5A. Then three important variables, sucrose, fish meal, and filling volume, were screened out by the Plackett-Burman (PB) design. The optimal level of these variables was further confirmed by response surface analysis. The final formulated medium was set as 35.2 g/L sucrose and 18.0 g/L fish meal, with filling volume of 34.6 mL, which could afford 440.3 mg/L production of B5A, approximately 4.4-fold higher than that in the original medium. The significantly improved productivity of B5A will facilitate the subsequent mechanistic and clinical studies of B5A.


[Neoadjuvant chemotherapy for primary breast cancer].
To evaluate the feasibility and effect of fluorouracil (5-Fu) in association with anthracycline-based regimen as neoadjuvant chemotherapy for primary breast cancer. For one hundred and eleven primary breast cancer patients with 114 lesions who were to be operated, two to six cycles of 5-Fu (continuous infusion) in association with epirubicin or pirarubicin and cyclophosphamide (CEFci or CTFci regimen) were given before operation. The response rate, side effect and its relation with tumor characteristics were studied. The overall response rate was 87.7%, of which the complete clinical response was 39.5%, pathological complete response was 23.7%, only one patient (0.9%) showed progressive disease. The regimen containing pirarubicin was superior to epirubicin regimen in pathological complete response rate (P < 0.05). Alopecia was mild in pirarubicin regimen as compared with epirubicin regimen but neutropenia was more severe in pirarubicin regimen than that in epirubicin regimen. Hormonal receptor expressions were significantly related to treatment response, the pathological complete response rate was 33.3% in oestrogen or progestin receptor negative tumors, but it was 7.5% in the positive tumors (P < 0.005). No correlation was observed between treatment response and tumor size, as well as HER-2 expression. CTFci/CEFci regimen as neoadjuvant chemotherapy is effective and safe for primary breast cancer. CTFci regimen is superior to CEFci regimen in response rate. The patients with negative hormonal receptor are more sensitive to the neoadjuvant chemotherapy.


The conduction system in corrected transposition with situs inversus.
The location and course of the conduction system were investigated by serial section in two hearts with corrected transposition of the great arteries in situs inversus. In the first case, characterized by dextrocardia, the interventricular septum was intact, while in the second case with levocardia, a high ventricular septal defect was associated with pulmonary atresia. In both hearts, a regular posterior connecting AV node was present, left-sided in one case and right-sided in the other. The bundle branches were distributed in the morphologically appropriate ventricles. Extensive hemorrhage and coagulation necrosis of the common bundle and bundle branches were observed in the first case, being consequent upon surgical replacement of the right-sided tricuspid valve. The common bundle ran below the membranous septum in this case. In the heart with ventricular septal defect it was related to the postero-inferior rim of the defect. In both cases, an accessory AV node was located anteriorly in the interatrial septum but did not connect with the ventricular musculature. The different position of the atrioventricular conducting tissue in corrected transposition, between cases in situs solitus and situs inversus, is emphasized.


Clinical detection of acute vestibulocerebellar disorders.
The acute onset of vertigo, nystagmus and postural instability, without brain-stem signs, is commonly attributed to a disorder of the labyrinth, the vestibular, sensory end organ. Identical symptoms can occur, however, with discrete infarctions or hemorrhages involving the central vestibulocerebellum. Whereas acute labyrinthine disorders are usually benign and self-limited, vascular injuries of the cerebellum may produce swelling, compression of the brain stem and acute hydrocephalus one to four days after the onset of symptoms. Therefore it is important to accurately distinguish between labyrinthine and vestibulocerebellar disorders with the neurologic examination. Acute labyrinthine disease causes unidirectional nystagmus with past-pointing and falling in the opposite direction of the nystagmus, environmental vertigo in the same direction and suppression of the nystagmus with visual fixation. Disorders of the vestibulocerebellum do not produce this consistent pattern of findings.


[Utility and drawbacks of fludeoxyglucose-positron emission tomography in locally advanced esophageal cancer treated by neoadjuvant chemotherapy followed by esophagectomy].
In Japan, neoadjuvant chemotherapy followed by esophagectomy has become the standard treatment for resectable advanced esophageal cancer. However, there are few reports on the utility of fludeoxyglucose( FDG)-positron emission tomography (PET) in such cases so far. In this study, we analyzed the utility and drawbacks of FDG-PET in patients who underwent neoadjuvant chemotherapy followed by surgery. Thirty-seven patients with cStage II or III thoracic esophageal cancer who received 2 courses of combination chemotherapy with 5-fluorouracil and cisplatin followed by surgery were enrolled this study. The maximum standardized uptake value (SUVmax) in the main lesion significantly decreased after chemotherapy (38.6%), and there was a greater decrease in SUVmax in patients with downstaging. Following chemotherapy, the esophageal lesions were not detected by FDG-PET in 6 patients. However, in these patients, the histopathological diagnosis of the resected specimens revealed that the cancer cells persisted. The SUVmax increased after chemotherapy in 5 patients, and in 4 of 5 patients recurrent disease was observed. In conclusion, we should understand the characteristics of FDG-PET in esophageal cancer after neoadjuvant chemotherapy and use it precisely.


Avoiding off-target effects in electrical stimulation of the cervical vagus nerve: Neuroanatomical tracing techniques to study fascicular anatomy of the vagus nerve.
Vagus nerve stimulation (VNS) is a promising therapy for treatment of various conditions that are resistant to standard medication, such as heart failure, epilepsy, and depression. The vagus nerve is a complex nerve providing afferent and efferent innervation of the pharynx, larynx, heart, tracheobronchial tree and lungs, oesophagus, stomach, liver, pancreas, small intestine and proximal colon. It is therefore a prime target for intervention for VNS. Surprisingly, the fascicular organisation of the vagus nerve at the cervical level is still not well understood. This, along with the current stimulation techniques, results in the entire nerve being stimulated, which leads to unwanted off-target effects. Neuronal tracing is a promising method to delineate the organ-specific innervation by the vagus nerve, thereby providing valuable insight into the fascicular anatomy. In this review we discuss the current knowledge of vagus nerve anatomy and neuronal tracers used for mapping of its organ-specific projections in various species. Efferent vagal projections are a chain of two neurones (pre- and postganglionic), while afferent projections consist of only one pseudounipolar neurone with one branch terminating in the target organ/tissue directly and another in the brainstem. It would be feasible to retrogradely trace the afferent fibres from their respective visceral targets and identify them at the cervical level using non-transsynaptic neuronal tracers. Using this to create a map of the functional anatomical organisation of the vagus nerve will enable selective VNS ultimately allowing for the avoidance of the off-target effects and improving overall efficacy.


Barriers and Facilitators to Genetic Testing for Familial Hypercholesterolemia in the United States: A Review.
Familial Hypercholesterolemia (FH) is an underdiagnosed condition in the United States (US) and globally, affecting an estimated 1/250 individuals. It is a genetic risk factor for premature cardiovascular disease and is responsible for an estimated 600,000 to 1.2 million preventable vascular events. Studies show that FH genetic testing can identify a causal gene variant in 60 to 80% of clinically suspected FH cases. However, FH genetic testing is currently underutilized in clinical settings in the US despite clinical recommendations and evidence supporting its use. Reasons for underutilization are not well understood. We conducted a literature review in the PubMed/MEDLINE database and eight peer-reviewed journals. After filtering for and reviewing 2340 articles against our inclusion criteria, we included nine commentaries or expert opinions and eight empirical studies reported between January 2014 and March 2019 in our review. After applying the Consolidated Framework for Implementation Research (CFIR), we identified a total of 26 potential barriers and 15 potential facilitators (estimated barrier to facilitator ratio of 1.73). We further estimated ratios of potential barriers to facilitators for each CFIR domain (Characteristics of Intervention, Outer Setting, Inner Setting, Characteristics of Individuals, and Process). Findings derived from our systematic approach to the literature and calculations of estimated baseline ratios of barriers and facilitators can guide future research to understand FH genetic testing implementation in diverse clinical settings. Our systematic approach to the CFIR could also be used as a model to understand or compare barriers and facilitators to other evidence-based genetic testing processes in health care settings in the US and abroad.


Bioavailability of coated and uncoated ZnO nanoparticles to cucumber in soil with or without organic matter.
There is a gap of knowledge for the fate, effects and bioavailability of coated and uncoated ZnO nanoparticles (NPs) in soil. Moreover, little is known about the effects of soil properties on effects of NPs on plants. In this study, the availability ZnO NPs in two soils with different organic matter content (one treated with cow manure (CM) and the other as untreated) was compared with their bulk particles. Results showed that coated and uncoated ZnO NPs can be more bioaccessible than their bulk counterpart and despite their more positive effects at low concentration (< 100mgkg-1), they were more phytotoxic for plants compared to the bulk ZnO particles at high concentration (1000mgkg-1) in the soil untreated with CM. The concentration of 1000mgkg-1 of ZnO NPs, decreased shoot dry biomass (52%) in the soil untreated with CM but increased shoot dry biomass (35%) in CM-treated soil compared to their bulk counterpart. In general, plants in the CM-treated soil showed higher Zn concentration in their tissues compared with those in untreated soil. The difference in shoot Zn concentration between CM-treated and untreated soil for NPs treatments was more than bulk particles treatment. This different percentage at 100mgkg-1 of bulk particles was 20.6% and for coated and uncoated NPs were 37% and 32%, respectively. Generally, the distribution of ZnO among Zn fractions in soil (exchangeable, the metal bound to carbonates, Fe-Mn oxides, organic matter and silicate minerals and the residual fraction) changed based on applied Zn concentration, Zn source and soil organic matter content. The root tip deformation under high concentration of NPs (1000mgkg-1 treatment) was observed by light microscopy in plants at the soil untreated with CM. It seems that root tip deformation is one of the specific effects of NPs which in turn inhibits plant growth and nutrients uptake by root. The transmission electron microcopy image showed the aggregation of NPs inside the plant cytoplasm and their accumulation adjacent to the cell membrane.


Bevacizumab and ovarian cancer.
Bevacizumab is a recombinant humanized monoclonal IgG1 antibody that targets vascular endothelial growth factor-A, and is indicated in the treatment of various tumors (colon, lung, renal, and glioblastoma). It has been recently approved for the treatment of ovarian cancer in various countries. This review summarizes the activity and toxicity of bevacizumab in the treatment of ovarian cancer, both as single-agent drug and in combination with cytotoxic chemotherapy. As a single-agent drug, it has shown response rates of 16-21% in the treatment of recurrent ovarian cancer. Two phase III randomized trials have been published evaluating the addition of bevacizumab to standard chemotherapy as front-line treatment of advanced ovarian cancer. In addition, trials evaluating the combination with chemotherapy in recurrent ovarian cancer (platinum-sensitive and platinum-resistant disease) have also been reported. All these trials showed a statistically significant improvement in progression-free survival although no improvement in overall survival has been reported. The main adverse event is hypertension. Other serious, but uncommon adverse events include gastrointestinal perforation as well as renal and central nervous system toxicity.


Statistical analysis of quality of life with missing data in cancer clinical trials.
We summarize issues that arise when considering quality of life (QOL) data in cancer clinical trials, especially those related to missing data. We describe different types of missing data mechanisms, and discuss ways of assessing and testing missing data mechanisms. A section on presentation of study design and results describes how graphical displays can effectively document the extent of the missing data problem, as well as describe its impact on interpretation of results. Finally, we describe several different statistical methods used to analyse repeated measures, with an emphasis on their properties and their ability to adequately handle different types of missing data mechanisms. We make recommendations as to the most appropriate methods, and suggest important directions for future research.


Versatile Analysis of DNA-Biomolecule Interactions in Solution by Hydrodynamic Separation and Single Molecule Detection.
DNA can interact with a wide array of molecules with a range of binding affinities, stoichiometry, and size-scales. We present a sensitive, quantitative, and versatile platform for sensing and evaluating these diverse DNA-biomolecule interactions and DNA conformational changes in free solution. Single molecule free solution hydrodynamic separation utilizes differences in hydrodynamic mobility to separate bound DNA-biomolecule complexes from unbound DNA and determine the associated size change that results from binding. Single molecule detection enables highly quantitative analysis of the fraction of DNA in the bound and unbound state to characterize binding behavior including affinity, stoichiometry, and cooperativity. A stacked injection scheme increases throughput to enable practical analysis of DNA-biomolecule interactions using only picoliters of sample per measurement. To demonstrate analysis of DNA-protein interactions on a local scale, we investigate binding of the E. coli single stranded binding protein to two DNA oligos both individually and in direct competition. We show that stoichiometry and cooperativity is a function of DNA length and verify these differences in binding characteristics through direct competition. To demonstrate analysis of DNA-small molecule interactions and global conformational changes, we also assess DNA condensation with the polyamine spermidine. We use hydrodynamic mobility to evaluate the size of spermidine-condensed DNA and single molecule burst analysis to evaluate DNA packing within the condensed globules relative to free-coiled DNA. This platform thus presents a versatile tool capable of quantitative and sensitive evaluation of diverse biomolecular interactions, complex properties, and binding characteristics.


Staging and current treatments for endometrial cancer.
Patients with Stage I endometrial carcinoma need to be further subdivided into low or high risk for recurrence. Treatment consists of surgery, with or without pre- or postoperative radiation. The supposed advantages of preoperative radiation are questionable. Definitive treatment recommendations for Stage I can be made only for Stage IaG1, where surgery alone suffices. Stage II patients have a high survival (80-90%) if there is no spread beyond the uterus. Although doxorubicin is the most active chemotherapeutic agent, the response rate is low. Response to progesterone is high (30-40%), but complete responses of prolonged duration are few.


"Lesbians are not women": feminine and lesbian sensibilities in Harmony Hammond's late-1970s sculpture.
Harmony Hammond's wrapped fabric sculptures are placed in context of the theories of gender and sexuality that circulated among lesbian and straight feminists at the time they were made, the late 1970s. Hammond has cited in particular Monique Wittig's novels, such as The Lesbian Body, and her essays including "The Straight Mind" where Wittig concludes that the lesbian is not a woman. The critique to which Wittig's lesbian separatism has been subjected by Judith Butler in her consideration of the appeal and limitations of essentialism also applies to Hammond's art. Hammond's use of vaginal imagery was instrumental to visualizing a lesbian sensibility, but the proposition of such a sensibility established a new problematic: a new essential category. The article concludes that because Hammond's work was produced in the context of a complex set of discourses, lesbian, feminist, and aesthetic, it resisted reduction to a singular meaning. Her sculptures avoided the pitfall of substituting one essence for another, lesbian for feminine sensibility, but activated both. The sculptures effectively queered vaginal imagery: When Hammond used vaginal imagery to represent lesbian sensibility, she subverted the equation of sex and gender and the essentialist notion of feminine sensibility.


Contributions of phonological memory, language comprehension and hearing to the expressive language of adolescents and young adults with Down syndrome.
Expressive language constitutes a major challenge to the development of individuals with Down syndrome. This paper investigates the relationships between expressive language abilities, language comprehension and the deficits in verbal short-term memory and hearing which are also associated with the syndrome. Tests of nonverbal ability, expressive language, verbal short-term memory, visuo-spatial memory, language comprehension and hearing were administered. Phonological memory, measured by nonword repetition, was significantly correlated with expressive language abilities measured by MLU and sentence recall. Adjusting for word repetition skills did not reduce this correlation, suggesting that the relationship did not depend on the fact that both tests required spoken output. Hearing did not contribute significantly to expressive language scores of participants who provided an intelligible narrative. However, level of hearing loss as well as other language and memory measures did differentiate these participants from those who were unable to produce an intelligible narrative. Phonological memory was closely associated with the expressive language abilities of individuals with Down syndrome. Hearing loss appeared to be less closely related except that individuals with uncorrected mild to moderate hearing loss had difficulty with the narrative task. Further research is necessary to establish the nature of these relationships.


Distinct luminal-type mammary carcinomas arise from orthotopic Trp53-null mammary transplantation of juvenile versus adult mice.
Age and physiologic status, such as menopause, are risk factors for breast cancer. Less clear is what factors influence the diversity of breast cancer. In this study, we investigated the effect of host age on the distribution of tumor subtypes in mouse mammary chimera consisting of wild-type hosts and Trp53 nullizygous epithelium, which undergoes a high rate of neoplastic transformation. Wild-type mammary glands cleared of endogenous epithelium at 3 weeks of age were subsequently transplanted during puberty (5 weeks) or at maturation (10 weeks) with syngeneic Trp53-null mammary tissue fragments and monitored for one year. Tumors arose sooner from adult hosts (AH) compared with juvenile hosts (JH). However, compared with AH tumors, JH tumors grew several times faster, were more perfused, exhibited a two-fold higher mitotic index, and were more highly positive for insulin-like growth factor receptor phosphorylation. Most tumors in each setting were estrogen receptor (ER)-positive (80% JH vs. 70% AH), but JH tumors were significantly more ER-immunoreactive (P = 0.0001) than AH tumors. A differential expression signature (JvA) of juvenile versus adult tumors revealed a luminal transcriptional program. Centroids of the human homologs of JvA genes showed that JH tumors were more like luminal A tumors and AH tumors were more like luminal B tumors. Hierarchical clustering with the JvA human ortholog gene list segregated luminal A and luminal B breast cancers across datasets. These data support the notion that age-associated host physiology greatly influences the intrinsic subtype of breast cancer.


Central autonomic modulation of cardiac baroreflex by corticotropin-releasing factor.
The effects of intracerebroventricular administration of corticotropin-releasing factor (CRF) on baroreflex control of heart rate were studied in conscious, unrestrained rats. Stimulus-response curves relating mean arterial pressure (MAP) to pulse interval were constructed for assessment of baroreflex function. CRF, intracerebroventricularly injected, elicited dose-related reductions of resting pulse interval, response range, and reflex gain and elevations of resting arterial pressure and MAP50 (arterial pressure at midrange). Earlier peripheral blockade of cardiac vagal influences with atropine methyl nitrate greatly attenuated the effects of CRF on resting pulse interval, response range, and reflex gain. In contrast, earlier blockade of cardiac sympathetic influences with intravenous propranolol treatment did not alter the ability of CRF to reduce resting pulse interval, response range, and reflex gain. The central nervous system effects of CRF on cardiac baroreflex function were qualitatively similar to the actions of atropine methyl nitrate. It is concluded that CRF alters transmission in central nervous system pathways mediating baroreceptor-induced activation of cardiac vagal motor neurons.


Incorporating Fermentation into Undergraduate Laboratory Courses.
Laboratory courses in universities have a responsibility to introduce current research practices and trends in scientific research to adequately prepare students for work in the field. One such research practice gaining popularity in recent years is that of green chemistry. Since the 1960s, increasing concern over the release of toxic chemicals into the environment has led to a push for more environmentally responsible chemistry. A growing faction of chemists has begun to adopt methods to eliminate chemical waste and support green chemistry. Fermentation is an ideal technique to demonstrate environmentally sustainable chemistry in an undergraduate laboratory class. Fermentation of complex natural products, as opposed to traditional organic synthesis, is beneficial as it supports a number of principles of green chemistry; it is conducted at ambient temperature and pressure, uses inexpensive and innocuous materials, makes use of renewable resources, and does not require a fume hood. Skills implemented during fermentation can be easily taught to upper-level Chemistry and Biochemistry undergraduate students, who typically have limited exposure to complex natural products in their coursework. Such a course would be interdisciplinary in nature, incorporating fungal biology and metabolism as well as organic chemistry. Students would learn a variety of skills, including growth media selection and preparation, inoculation of fungal cultures, extraction of natural products, and purification and characterization of metabolites. Experiments of this nature would allow for discussions of several areas of research: green chemistry, natural products and their application to medicine, identification of functional groups in complex molecules by spectroscopy, and introduction to biochemistry and metabolism. Roquefortine C, a prenylated indole alkaloid readily produced by a variety of species of Penicillia, is an excellent candidate for demonstrating fermentation in a laboratory classroom setting, owing to its ease of purification from other metabolites and its unique structural features.


Effects of ingestion of collagen peptide on collagen fibrils and glycosaminoglycans in Achilles tendon.
In order to investigate whether the oral ingestion of collagen peptide affects the extracellular matrix of tendon, two doses (0.2 g/kg and 1.0 g/kg body weight) were orally administered daily for 56 d to a rabbit, and both the size of collagen fibrils and the amount of glycosaminoglycans in the Achilles tendon were measured in comparison with those in a rabbit fed with a control protein, lactalbumin, or water alone. Ingestion of collagen peptide or lactalbumin induced a significant increase in collagen fibril diameter and a decrease in fibril density except for a high dose of lactalbumin compared with the water control. A histogram pattern of fibril diameter in a high dose of collagen peptide showed a peak at 160-180 nm, which was not observed in other groups. However the percentage of diameters over 200 nm was the lowest in this group but highest in the low-dose group of collagen peptide. The mean fibril diameter and mass average diameter of a high dose of collagen peptide were significantly smaller than those in a low dose. The amount of dermatan sulphate increased in the high-dose groups, while the amount of hyaluronic acid decreased in rabbits fed with collagen peptide or lactalbumin at either dose. These results suggest that the ingestion of collagen peptide affects the size of collagen fibrils and composition of glycosaminoglycans in the Achilles tendon and thus may improve the mechanical properties of the Achilles tendon.


[Determination of the gestational age in late pregnancy by sonography (author's transl)].
In 219 normal pregnancies between 28 and 42 weeks the relationships of different ultrasound measurements of the fetal head and the fetal body to the gestational age were determined in order to improve the accurate diagnosis of fetal maturity during the last trimester of pregnancy. With the A and B ultrasound apparatus of Kretz, the biparietal diameter was determined by the A mode, the fronto occipital plane and the largest abdominal diamter were determined by the B scan. The following ultrasound measurements were taken: the two horizontal and transverse diameters, the circumferences and the planes. The abdominal measurements showed a higher rate of growth, compared to the cranial measurements a lesser rate of growth retardation and a higher variation of the normals. The correlation of the abdominal measurements with the gestational age was worse than that of the cranial measurements. Wth the exception of the biparietal diameter in the A mode, the mean diameters of the cranial and abdominal measurements gave better results than the single diameters. Circumference and surface measurements showed no advantages. The best results were obtained by combined fetal head and body measurements using the summation of the biparietal diameter in the A mode and the mean abdominal diameter in the B scan.


In vitro immunoglobulin isotype suppression in immunoglobulin A nephropathy.
IgA nephropathy (IgAN) is characterized by mesangial IgA deposition, and up-regulation of the IgA system is frequently described. This study investigated in vitro immunoglobulin isotype production by peripheral blood mononuclear cells from patients with IgAN and controls, without mitogenic stimulation and under the influence of cell cycle inhibitors and cyclosporin A (CyA). In controls, only IgM production was suppressed by cell cycle inhibitors, and no isotype was affected by CyA. This suppressible IgM production may represent antigen-independent 'natural antibody'. In IgAN, IgM, IgA and IgG were all suppressed by cell cycle inhibitors, and CyA suppressed IgA production. This state of altered immune activation in IgAN demonstrates T-cell-independent IgA production, suggesting that it is IgA 'natural antibody'. In conclusion, we suggest that mesangial IgA deposition may not be antigen dependent but due to some non-immune characteristic of the IgA molecule.


Cardiac involvement in patients with primary biliary cholangitis: A 14-year longitudinal survey-based study.
Patients with primary biliary cholangitis (PBC) can have extrahepatic manifestations. However, data about cardiac involvement of PBC is limited. We aimed in this study to analyze the clinical characteristics in patients with PBC complicated with and without cardiac involvement, and the risk factors of cardiac involvement in PBC. PBC patients admitted to Peking Union Medical College Hospital between January 2002 and February 2016 were consecutively enrolled. Structured interview, systemic rheumatologic examination, and laboratory tests were conducted for each patient, and risk factors of cardiac involvement were analyzed by comparing patients with and without cardiac involvement. In total, 580 PBC patients were enrolled, and cardiac involvement was identified in 24 patients (4%), with 11 male (46%) and a mean age of 57±8 year. Cardiomyopathy and arrhythmias were presented in 17 (70.8%) and 21 (87.5%) patients, respectively. Patients with cardiac involvement were more frequently male (46% vs. 11%, P<0.01), complicated with inflammatory myopathy (IM) (58% vs. 1%, P<0.01), and had a longer disease course (median, 72 vs 24 month, P<0.01). Furthermore, concomitant IM was the independent risk factor of cardiac involvement in PBC (OR = 77.333, 95% CI: 23.704-252.294). Cardiac involvement was a rare complication of PBC, which was more frequently observed in male or long-course patients. Importantly, concomitant IM was the strong independent risk factor of cardiac involvement in PBC. Given cardiac involvement is a serious complication, thorough evaluation of cardiac manifestation in high-risk PBC patients is highly recommended.


Personnel administration: personnel problems in small medical libraries.
The shortage of medical librarians is felt most acutely by the small library, because the librarian's job may not seem challenging or the parent institution or organization is not willing or able to pay a competitive salary. The librarian's position often is filled, therefore, by an unqualified person, or a single professional librarian may have nonprofessionals as assistants. This paper discusses cooperative arrangements for sharing the service of a professional librarian and enumerates means of providing instruction for nonprofessional personnel.


Gene Structure Analysis of Chemokines and Their Receptors in Allotetraploid Frog, Xenopus laevis.
Chemokines, relatively small secreted proteins, are involved in cell migration and function in various biological events, including immunity, morphogenesis, and disease. Due to their nature, chemokines tend to be a target of hijacking of immunity by virus and therefore show an exceptionally high mutation rate. Xenopus laevis is considered an excellent model to investigate the effect of whole-genome duplication for gene family evolution. Because its allotetraploidization occurred around 17-18 million years ago, ancestral subgenomes L and S were well conserved. Based on the gene model of human and diploid frog Xenopus tropicalis, we identified 52 chemokine genes and 26 chemokine receptors in X. laevis. The retention rate of the gene in the X. laevis L and S subgenomes was 96% (45/47) and 68% (32/47), respectively. We conducted molecular phylogenetic analysis and found clear orthologies in all receptor genes but not in the ligand genes, suggesting rapid divergences of the ligand. dN/dS calculation demonstrated that dN/dS ratio greater than one was observed in the four ligand genes, cxcl8b.1.S, cxcl18.S, ccl21.S, and xcl1.L, but nothing in receptor genes. These results revealed that the whole-genome duplication promotes diversification of chemokine ligands in X. laevis while conserving the genes necessary for homeostasis, suggesting that selective pressure also supports a rapid divergence of the chemokines in amphibians.


A red squirrel associated adenovirus identified by a combined microarray and deep sequencing approach.
In 2013, an adult red squirrel (Sciurus vulgaris) was found dead showing, during necropsy, acute diffuse catarrhal enteritis. Using a virus screening pipeline based on a pan-viral DNA microarray with downstream next-generation sequencing a novel squirrel-associated virus was identified with mastadenovirus-typical sequence elements. Phylogenetic analysis of hexon protein amino acid sequences demonstrated the highest similarity to Equine adenovirus (AdV) 2, but a strong divergence to a squirrel AdV from Korea and other rodent AdVs. Shorter hexon gene segment investigations confirmed a close relationship with other squirrel-derived AdVs from Europe. The novel virus species was tentatively designated as Squirrel Adenovirus-1.


Somatic awareness and symptom attribution in ischemic stroke patients.
Patient delay in seeking treatment for stroke symptoms is a major factor in morbidity and mortality due to stroke. Every 4 minutes, someone dies of a stroke, and 54% of those deaths occur before the patient reaches the hospital. The aim of this study was to determine the relationship of somatic awareness and symptom attribution to treatment-seeking behaviors in ischemic stroke patients. This was a descriptive, correlational pilot study. This study was carried out in three tertiary-care hospitals in Philadelphia, Pennsylvania, and Binghamton, New York. Twenty patients diagnosed with ischemic stroke were included. The Modified Somatic Perception Questionnaire measured somatic awareness, and the Symptom Interpretation Questionnaire measured symptom attribution. There was no relationship identified between somatic awareness or symptom attribution and treatment-seeking behaviors. Significance was identified, using t-test comparisons, between physical attribution scores and insurance status (p = .038) and family history of stroke (p = .026). Significant correlations, r = .386 and r = .433, respectively, were identified between somatic awareness and symptom attribution and education. Somatic awareness and symptom attribution were not found to be significantly correlated to time to care. These results must be tempered with the knowledgethat the data were derived from a small sample and that some data were missing due to a lack of recall. Significance was identified, in a comparison of the group means, involving insurance status and family history of stroke, to somatic awareness and symptom attribution. Significant correlation was identified between education and these factors. Future research involving these factors, using a revised methodology, should be considered.


Genomic organization of Drosophila poly(ADP-ribose) polymerase and distribution of its mRNA during development.
Poly(ADP-ribosyl)ation of proteins catalyzed by poly(ADP-ribose) polymerase (PARP; EC 184.108.40.206) modulates several biological activities. However, little is known about the role of PARP in developmental process. Here we report the organization of the Drosophila PARP gene and the expression patterns during Drosophila development. The Drosophila PARP gene was a single copy gene mapped at 81F and composed of six exons. Organization of exons corresponds to the functional domains of PARP. The DNA-binding domain was encoded by exons 1, 2, 3, and 4. The auto-modification domain was encoded by exon 5, and the catalytic domain was in exon 6. The promoter region of the PARP gene contained putative TATA box and CCAAT box unlike human PARP. Expression of the PARP gene was at high levels in embryos at 0-6 h after egg laying and gradually decreased until 8 h. PARP mRNA increased again at 8-12 h and was observed in pupae and adult flies but not in larvae. In situ mRNA hybridization of embryos revealed large amount of PARP mRNA observed homogeneously except the pole cells at the early stage of embryos, possibly due to presence of the maternal mRNA for PARP, and decreased gradually until the stage 12 in which stage PARP mRNA localized in anal plates. At late stage of embryogenesis PARP mRNA was localized in cells along the central nervous system.


Nasal irrigation reduces postirradiation rhinosinusitis in patients with nasopharyngeal carcinoma.
Nasopharyngeal carcinoma (NPC) is a common head and neck cancer in Taiwan. Radiotherapy (RT) is the standard treatment for NPC. The newer RT technique, intensity-modulated radiotherapy (IMRT) has become popular in recent years. IMRT-induced rhinosinusitis is not uncommon in postirradiated NPC patients but the incidence and the disease course have not been reported. The purpose of this study was to determine the IMRT effect on the paranasal sinuses and evaluate the efficacy of nasal irrigation on the management of RT-induced rhinosinusitis. NPC patients who completed IMRT from October 2004 to May 2006 were enrolled in the study and were randomly allocated to irrigation or nonirrigation groups. Patients in the irrigation group performed daily nasal irrigation until 6 months after RT. The severity of postirradiated rhinosinusitis was evaluated by nasal endoscopy, questionnaire, and computed tomography until a year after RT. One hundred seven postirradiated NPC patients completed the study. Among them, 44 patients performed daily nasal irrigation until 6 months after RT, and the other 63 patients did not perform nasal irrigation after RT. Patients in the irrigation group had significantly lower endoscopic and questionnaire scores than patients in the nonirrigation group (p = 0.001 and 0.0001, respectively) from pre-RT to 6 months after RT. The between-group differences were most obvious at the post-RT second and third months. Rhinosinusitis is a common acute post-RT complication in NPC patients. Our results showed that nasal irrigation was a safe and effective method for the management of this acute complication.


Postoperative radiotherapy in squamous cell carcinoma of the oral cavity: the importance of the overall treatment time.
To test the hypothesis that (1) the distinction between intermediate- and high-risk patients by clustering different prognostic factors results in a significant difference in treatment outcome and (2) a shorter interval between surgery and radiotherapy and shorter overall treatment times of radiation (OTTRT) result in higher rates of locoregional control (LRC). Included were patients (n = 217) with previously untreated squamous cell carcinoma of the oral cavity treated with radical surgery and postoperative radiotherapy. Patients with extranodal spread or microscopic residual disease and patients with two or more other risk factors (i.e., N2b-N3, >1 nodal level involved, perineural growth, or stage T3-T4) were classified as high-risk patients. Patients with only one other risk factor were classified as intermediate risk. In the intermediate-risk group, the 3-year LRC was 87% as compared with 66% in the high-risk group (p = 0.0005). No association was found between interval and LRC. However, the OTTRT was significantly associated with LRC. The 3-year LRC was 87%, 75%, 69%, and 51% when the OTT was <6 weeks, 6-7 weeks, 7-8 weeks, and >8 weeks, respectively (p = 0.0004). The 3-year overall survival (OS) in the intermediate risk patients was 74% compared with 50% in the high-risk group (p = 0.0014). A significant association was also found between the OS and OTTRT. The OS increased from 50% when the OTTRT was >8 weeks to 74% when the OTT was <6 weeks (p = 0.006). Similar results were found with regard to the disease-free survival (DFS). In the multivariate analysis, both risk group and OTT were significantly associated with LRC, DFS, and OS. No significant interaction term was present between these two factors, which means that the OTT was of importance both for the high-risk and the intermediate-risk patients. In the subset of patients with carcinoma of the oral cavity, the classification of high- and intermediate-risk patients by clustering a number of prognostic factors provides important prognostic information regarding LRC, DFS, and OS. The OTT was the most important prognostic factor both in the high-risk and intermediate-risk patients. Reducing the OTT to 6 weeks or less is a rather simple measure to achieve a considerable improvement of the outcome of treatment in this category of patients.


A potent mechanism-inspired O-GlcNAcase inhibitor that blocks phosphorylation of tau in vivo.
Pathological hyperphosphorylation of the microtubule-associated protein tau is characteristic of Alzheimer's disease (AD) and the associated tauopathies. The reciprocal relationship between phosphorylation and O-GlcNAc modification of tau and reductions in O-GlcNAc levels on tau in AD brain offers motivation for the generation of potent and selective inhibitors that can effectively enhance O-GlcNAc in vertebrate brain. We describe the rational design and synthesis of such an inhibitor (thiamet-G, K(i) = 21 nM; 1) of human O-GlcNAcase. Thiamet-G decreased phosphorylation of tau in PC-12 cells at pathologically relevant sites including Thr231 and Ser396. Thiamet-G also efficiently reduced phosphorylation of tau at Thr231, Ser396 and Ser422 in both rat cortex and hippocampus, which reveals the rapid and dynamic relationship between O-GlcNAc and phosphorylation of tau in vivo. We anticipate that thiamet-G will find wide use in probing the functional role of O-GlcNAc in vertebrate brain, and it may also offer a route to blocking pathological hyperphosphorylation of tau in AD.


Scalar localization by computed tomography of cochlear implant electrode carriers designed for deep insertion.
This study aimed to evaluate the possibility of predicting radiologically the scalar localization of a 31.5-mm-long, free-fitting electrode carrier for cochlear implantation, using conventional planar computed tomography. A cross-sectional human temporal bone study was conducted. Twenty human temporal bones were acquired postmortem and implanted with 31.5-mm-long electrode carriers. Ten of these were implanted into the scala tympani using the round window approach, whereas the other 10 electrodes were inserted into the scala vestibuli by cochleostomy. Computed tomography was then performed, and 2 experienced blinded radiologists evaluated the intracochlear position of the array. The estimated position of the electrode carrier was described using a 5-point scale. After sectioning and histologic investigation, the results of the radiologic and histologic investigations were compared. In 17 of 20 cases, it was possible to estimate the correct position of the electrode carrier within the basal turn of the cochlea by means of computed tomography. As the insertion angles widened beyond 360 degrees, it became increasing difficult for the radiologists to correctly determine the position of the electrode carrier. The comparison of our temporal bone experiment results with the computed tomography results revealed the difficulty of assessing the correct position of intracochlear electrodes. Scalar localization of deeply inserted electrode carriers cannot be precisely determined by means of computed tomography.


Long-term follow-up and prognostic factors for advanced thymic carcinoma.
The aim of this study was to evaluate the long-term survival outcomes in patients with advanced thymic carcinoma and identify prognostic factors influencing the survival. We retrospectively analyzed 90 consecutive patients with pathologically confirmed advanced thymic carcinoma (Masaoka III and IV) in our institute, from December 2000 to 2012. Age, sex, clinical characteristics, laboratory findings, Masaoka and tumor node metastasis staging, pathologic grade, and treatment modalities were analyzed to identify prognostic factors associated with the progress-free survival (PFS) and the overall survival (OS) rates. Statistical analysis was conducted using SPSS, version 19.0 (SPSS, Inc, Chicago, IL). A total of 73 (81.1%) male and 17 (18.9%) female patients participated in the study. The median follow-up time was 75 months (range, 20-158 months). The 5-year PFS and OS rates were 23.6% (95% confidence interval [CI], 14.6%-33.8%) and 35.7% (95% CI, 25.1%-46.4%), respectively. The multivariate Cox regression model analysis showed that factors improving the PFS were the normal lactate dehydrogenase (LDH) level (P<0.001), Masaoka III stage (P=0.028), and radiotherapy (RT) (P<0.001). The LDH (P<0.001), T stage (P<0.001), and the pathologic grade (P=0.047) were independently prognostic of OS. Long-term follow-up of the advanced thymic carcinoma showed poor outcomes of PFS and OS. LDH, Masaoka stage, and RT affected the PFS, and LDH, T stage, and pathologic grade seemed to affect the OS. Establishing a better staging system for predicting outcomes would be warranted.


Assessment of ATP-Bioluminescence and Dipslide Sampling to Determine the Efficacy of Slaughterhouse Cleaning and Disinfection Compared with Total Aerobic and Enterobacterales Counts.
Inadequate cleaning and disinfection (C&D) in slaughterhouses can cause bacterial contamination of meat, resulting in foodborne disease and reduced meat quality. Different methods for monitoring the efficacy of C&D procedures are available, but few studies have assessed their reliability. This study examined C&D efficacy in slaughterhouses and evaluated the diagnostic performance of methods for measuring surface hygiene. One red meat and one poultry slaughterhouse in Sweden were each visited on six occasions before and six occasions after C&D. Sampling points were sampled with: swabbing and plating for total aerobic bacteria (TAB) and Enterobacterales (EB); dipslides for total viable count; and ATP-bioluminescence tests. To evaluate the diagnostic performance of the dipslide and ATP-bioluminescence methods, the results were compared with (TAB) as a reference. In total, 626 samples were collected. For the majority of samples, TAB was lower after than before C&D and EB were mainly detected before C&D, indicating C&D efficacy. Greater reductions in mean TAB were observed in processing areas (2.2 and 2.8 log CFU/100 cm2 in red meat and poultry slaughterhouse, respectively) than in slaughter areas (1.3 log CFU/100 cm2 in both slaughterhouses). Approximately half of all samples were assessed as non acceptably clean (52% for red meat and 46% for poultry slaughterhouse) according to previously published thresholds. Critical food contact surfaces that were insufficiently cleaned and disinfected were plucking fingers, shackles, and a post-dehairing table. Cleaning and disinfection of drains and floors were inadequate. The ATP-bioluminescence method showed low specificity compared with the reference (TAB) in both the red meat (0.30) and poultry slaughterhouses (0.64). The sensitivity of dipslides was low (0.26) in the red meat slaughterhouse compared with TAB. A combination of ATP-bioluminescence and dipslides could provide more accurate estimates of C&D efficacy.


The role of the entorhinal cortex in extinction: influences of aging.
The entorhinal cortex is perhaps the area of the brain in which neurofibrillary tangles and amyloid plaques are first detectable in old age with or without mild cognitive impairment, and very particularly in Alzheimer's disease. It plays a key role in memory formation, retrieval, and extinction, as part of circuits that include the hippocampus, the amygdaloid nucleus, and several regions of the neocortex, in particular of the prefrontal cortex. Lesions or biochemical impairments of the entorhinal cortex hinder extinction. Microinfusion experiments have shown that glutamate NMDA receptors, calcium and calmodulin-dependent protein kinase II, and protein synthesis in the entorhinal cortex are involved in and required for extinction. Aging also hinders extinction; it is possible that its effect may be in part mediated by the entorhinal cortex.


An allelic variant of the PmrB sensor kinase responsible for colistin resistance in an Escherichia coli strain of clinical origin.
We investigated the colistin resistance mechanism in an Escherichia coli strain (LC711/14) isolated in Italy in 2014, from an urinary tract infection, which was previously shown to express a colistin resistance mechanism different from mcr-1. LC711/14 was found to carry a novel mutation in the pmrB gene, resulting in a leucine to proline amino acid substitution at position 10 of the PmrB sensor kinase component of the PmrAB signal transduction system. The role of this substitution in colistin resistance was documented by expression of the wild-type and mutated alleles in a pmrB deletion derivative of the E. coli reference strain MG1655, in which expression of the mutated allele conferred colistin resistance and upregulation of the endogenous pmrHFIJKLM lipid A modification system. Complementation of LC711/14 with the wild-type pmrB allele restored colistin susceptibility and decreased expression of pmrHFIJKLM, confirming the role of this PmrB mutation. Substitution of leucine at position 10 of PmrB with other amino acids (glycine and glutamine) resulted in loss of function, underscoring a key role of this residue which is located in the cytoplasmic secretion domain of the protein. This work demonstrated that mutation in this domain of the PmrB sensor kinase can be responsible for acquired colistin resistance in E. coli strains of clinical origin.


Normalization of red cell enolase level following allogeneic bone marrow transplantation in a child with Diamond-Blackfan anemia.
We describe a girl with Diamond-Blackfan anemia with accompanying red cell enolase deficiency. At the age of 9 yr old, the patient received allogeneic bone marrow transplantation from her HLA-identical sister who had normal red cell enolase activity. While the post transplant DNA analysis with short tandem repeat has continuously demonstrated a stable mixed chimerism on follow-up, the patient remains transfusion independent and continues to show a steady increase in red cell enolase activity for over two and a half years following bone marrow transplantation.


Soft, flexible pressure sensors for pressure monitoring under large hydrostatic pressure and harsh ocean environments.
Traditional rigid ocean pressure sensors typically require protection from bulky pressure chambers and complex seals to survive the large hydrostatic pressure and harsh ocean environment. Here, we introduce soft, flexible pressure sensors that can eliminate such a need and measure a wide range of hydrostatic pressures (0.1 MPa to 15 MPa) in environments that mimic the ocean, achieving small size, high flexibility, and potentially low power consumption. The sensors are fabricated from lithographically patterned gold thin films (100 nm thick) encapsulated with a soft Parylene C film and tested in a customized pressure vessel under well-controlled pressure and temperature conditions. Using a rectangular pressure sensor as an example, the resistance of the sensor is found to decrease linearly with the increase of the hydrostatic pressure from 0.1 MPa to 15 MPa. Finite element analysis (FEA) reveals the strain distributions in the pressure sensor under hydrostatic pressures of up to 15 MPa. The effect of geometry on sensor performance is also studied, and radially symmetric pressure sensors (like circular and spike-shaped) are shown to have more uniform strain distributions under large hydrostatic pressures and, therefore, have a potentially enhanced pressure measurement range. Pressure sensors of all geometries show high consistency and negligible hysteresis over 15 cyclic tests. In addition, the sensors exhibit excellent flexibility and operate reliably under a hydrostatic pressure of 10 MPa for up to 70 days. The developed soft pressure sensors are promising for integration with many platforms including animal tags, diver equipment, and soft underwater robotics.


Myocardial Surgical Revascularization in Patients with Reduced Left Ventricular Ejection Fraction.
Myocardial surgical revascularization in patients with low left ventricular ejection fraction (LVEF) is accompanied by a high rate of morbidity and mortality. The aim of this study was to investigate and eliminate the reasons for the most common perioperative and postoperative complications. A total of 64 were analyzed. of patients during 2019 who underwent coronary artery bypass grafting (CABG), average age 61.29±9.12 years. Out of the total number of operated patients, there were 16 women and 48 men. Patients were divided into two groups. The first group consisted of patients who underwent surgery with the use of cardiopulmonary bypass (cCABG-CPB) and the second group those who underwent surgery without the use of cardiopulmonary bypass (OPCAB). In 41 patients, myocardial infarction was previously recorded. Critical stenosis of the main trunk of the left coronary artery was present in 14 patients. The incidence of postoperative complications was higher in the cCABG-CPB 16/10 group (p0.030). In our study, we confirmed that myocardial revascularization is justified, especially in the case of multivessel coronary disease. In the long term, it significantly improves the systolic function of the left ventricle, and thus and quality and length of life.