Time lapse video microscopy using an animation control unit.
A system for time lapse video microscopy incorporating an edit recorder and animation control unit is described, allowing both colour recording, forward and reverse playback without synchrony bars, and the identification of each individual frame recorded. This is achieved by using the edit recorder, controlled by the animation unit to produce continuous series of insert edits using the preroll facility of the recorder. A feature of this system is the preview of the last hour's recording which is constantly played as the preroll of the edit recorder. Tissue culture flasks modified for stable focus are also described.


Dynamic analysis of iris changes and a deep learning system for automated angle-closure classification based on AS-OCT videos.
To study the association between dynamic iris change and primary angle-closure disease (PACD) with anterior segment optical coherence tomography (AS-OCT) videos and develop an automated deep learning system for angle-closure screening as well as validate its performance. A total of 369 AS-OCT videos (19,940 frames)-159 angle-closure subjects and 210 normal controls (two datasets using different AS-OCT capturing devices)-were included. The correlation between iris changes (pupil constriction) and PACD was analyzed based on dynamic clinical parameters (pupil diameter) under the guidance of a senior ophthalmologist. A temporal network was then developed to learn discriminative temporal features from the videos. The datasets were randomly split into training, and test sets and fivefold stratified cross-validation were used to evaluate the performance. For dynamic clinical parameter evaluation, the mean velocity of pupil constriction (VPC) was significantly lower in angle-closure eyes (0.470 mm/s) than in normal eyes (0.571 mm/s) (P < 0.001), as was the acceleration of pupil constriction (APC, 3.512 mm/s2 vs. 5.256 mm/s2; P < 0.001). For our temporal network, the areas under the curve of the system using AS-OCT images, original AS-OCT videos, and aligned AS-OCT videos were 0.766 (95% CI: 0.610-0.923) vs. 0.820 (95% CI: 0.680-0.961) vs. 0.905 (95% CI: 0.802-1.000) (for Casia dataset) and 0.767 (95% CI: 0.620-0.914) vs. 0.837 (95% CI: 0.713-0.961) vs. 0.919 (95% CI: 0.831-1.000) (for Zeiss dataset). The results showed, comparatively, that the iris of angle-closure eyes stretches less in response to illumination than in normal eyes. Furthermore, the dynamic feature of iris motion could assist in angle-closure classification.


Exploring nurses' intention to use a computerized platform in the resuscitation unit: development and validation of a questionnaire based on the theory of planned behavior.
In emergency department resuscitation units, writing down information related to interventions, physical examination, vital signs, investigations, and treatments ordered is a crucial task carried out by nurses. To facilitate this task, a team composed of emergency physicians, nurses, and one computer engineer created a novel electronic platform equipped with a tactile screen that allows systematic collection of critical data. This electronic platform also has medical software (ReaScribe+) that functions as an electronic medical record and a clinical decision support system. To develop and validate a questionnaire that can help evaluate nurses' intention to use a novel computerized platform in an emergency department resuscitation unit, based on Ajzen's theory of planned behavior (TPB). The sample for this study was composed of 87 nurses who worked in the resuscitation unit of a tertiary trauma center. We held three focus groups with nurses working in the resuscitation unit to identify the salient modal beliefs regarding their intended use of a new electronic medical charting system for the care of trauma patients. The system included a clinical decision support tool. We developed a questionnaire in which salient modal beliefs were used as items to evaluate the TPB constructs. We also added 13 questions to evaluate nurses' computer literacy. The final questionnaire was composed of 46 questions to be answered on a 7-point Likert scale. All nurses in the resuscitation unit and present during a regular work shift were individually contacted by the principal investigator or a research assistant (phase 1). A subsample of the nurses who completed the questionnaire was invited to complete it a second time 2 weeks later (phase 2). In phase 1, we received 62 of the 70 questionnaires administered (89% response rate). Of the 27 questionnaires administered in phase 2 (retest phase), 25 were completed (93% response rate). The questionnaire showed very good internal consistency, as Cronbach alpha was higher than .7 for all constructs. Temporal stability was acceptable with intraclass correlations between .41 and .66. The intention to use the electronic platform to chart the resuscitation of trauma patients was very high among the respondents. In the logistic regression model, the only construct that predicted nurses' intention to adopt the computerized platform was the professional norm (odds ratio 3.31, 95% confidence interval 1.41-7.78). We developed and validated a questionnaire that can now be used in other emergency departments prior to implementation of the computerized platform. The intention to adopt was very high among the respondents, which suggests that the implementation of this innovation could be successful at our institution.


Quality improvement and patient care checklists in intrahospital transfers involving pediatric surgery patients.
Intrahospital transfers are necessary but hazardous aspects of pediatric surgical care. Plan-Do-Study-Act processes identify risks during hospitalization and improve care systems and patient safety. A multidisciplinary team developed a checklist that documented patient data and handoffs for all intrahospital transfers involving pediatric surgical inpatients. The checklist summarized major clinical events and provided concurrent summaries by 3-month quarters (Q) over 1 year. There were 903 intrahospital transfers involving 583 inpatients undergoing surgery. Total handoffs were documented in 436 (75% of 583), with greater than 1 handoff in 202 (46% of 436). Documented problems occurred in 31 transfers (3.4%), the most during Q1 (19/191; 9.9%). Incidence fell to 3.5% (9/260) in Q2, 0.4% (1/243) in Q3, and 1.0% (2/209) in Q4 (P < .001). Patient care issues (14/31; 45%) were most common, followed by documentation (10, 32%) and process problems (7, 23%). The quality improvement team was able to resolve patient instability during transport (5 in Q1, none in Q3, Q4) and poor pain control (3 in Q2, 1 in Q3, Q4). Of the patients, 3.2% had identified problems with patient care during intrahospital transfer. Plan-Do-Study-Act review emphasizes ongoing process analysis by multidisciplinary teams. Checklists reinforce communication and provide feedback on whether system goals are being achieved.


Characterization of three mutations causing von Willebrand disease type IIA in five unrelated families.
Von Willebrand disease (vWD) type IIA is characterized by decreased ristocetin-induced platelet aggregation, and by the absence from plasma of high molecular weight multimers of von Willebrand factor (vWF). Most mutations causing vWD type IIA are clustered within the A2 domain of the mature vWF subunit that is encoded by exon 28. Using the polymerase chain reaction (PCR), the entire exon 28 from patients with vWD type IIA and normal controls was amplified and sequenced. Three missense mutations were detected that result in the amino acid substitutions were detected that result in the amino acid substitutions Arg(834)----Trp, Gly(742)----Glu, and Ser(743)----Leu. The first mutation occurred independently in three unrelated families; each of the latter mutations was found in one family. By restriction endonuclease analysis and allele-specific oligonucleotide (ASO) hybridization the mutations were confirmed in affected family members and excluded in unaffected members and 50 normal controls. The apparently high frequency of identical independent mutations among patients with vWD type IIA suggests that a precise diagnosis may be possible in a majority of patients using relatively simple recombinant DNA screening assays.


Head and neck squamous cell carcinoma: Exploring frontiers of combinatorial approaches with tyrosine kinase inhibitors and immune checkpoint therapy.
Squamous cell carcinomas (SCC) are the most common malignant tumors that arise in the head and neck. Despite advances in the management of affected patients, the mortality burden of these tumors is increasing every year. The discovery of a vast genetic landscape has revealed new opportunities for therapeutic intervention of head and neck SCC (HNSCC). Molecular alterations of tyrosine kinases are involved in the pathogenesis of cancer and may help keep cancer cells from growing. Currently, many drugs inhibit this enzyme family and are being studied by the pharmaceutical industry opening the room to expand the use and efficacy of this therapeutic modality alone or using combinatorial approaches including checkpoint inhibitors for treatment. In this paper, we explored the role of tyrosine kinases inhibitors of HNSCC, and clinical trials related to these molecules, expecting to provide references for HNSCC therapy.


The role of global top-down factors in local eye-movement control in reading.
Although the development of the field of reading has been impressive, there are a number of issues that still require much more attention. One of these concerns the variability of skilled reading within the individual. This paper explores the topic in three ways: (1) it quantifies the extent to which, two factors, the specific reading task (comprehension vs. word verification) and the format of reading material (sentence vs. passage) influence the temporal aspects of reading as expressed in word-viewing durations; (2) it examines whether they also affect visuomotor aspects of eye-movement control; and (3) determine whether they can modulate local lexical processing. The results reveal reading as a dynamic, interactive process involving semi-autonomous modules, with top-down influences clearly evident in the eye-movement record.


Ferritin and lysozyme distribution in normal and abnormal duodenal mucosae.
The distribution of ferritin and lysozyme in 19 normal and abnormal duodenal biopsies was studied by an immunoperoxidase technique. The abnormal biopsies included cases of chronic duodenitis with gastric metaplasia, gastric heterotopia, villous atrophy, and a case of hemochromatosis. Ferritin is demonstrated in duodenal absorptive cells, with the staining being most intense in the hemochromatosis case. It was absent in duodenal cells showing gastric metaplasia and in the surface epithelial cells of most biopsies with villous atrophy and gastric heterotopia. Lysozyme-positive mononuclear inflammatory cells were markedly increased in all abnormal biopsies. Not all lysozyme-positive cells were ferritin positive. The latter were especially abundant in areas with gastric metaplasia. It is suggested that this abundance may be related to passive diffusion of intestinal contents, particularly iron, through the metaplastic areas, and consequently there may be a relationship between the presence of duodenal gastric metaplasia and uncontrolled iron absorption.


Factors that influence the outcome of pregnancy in middle-class women.
A retrospective analysis was made of 1,080 singleton pregnancies of middle-class women to identify factors that influence outcome as measured by birth weight, gestational age, Apgar scores, and the incidence of complications. Increased birth weight was associated with increases in weight gain, maternal age, gestational age, maternal initial body weight, maternal hemoglobin and hematocrit values at 3 months, hemoglobin levels at 7 months, and socioeconomic status. For women whose initial weights were 90% to 135% of the standard, highest birth weights occurred when weight gains exceeded 29 lb. A 1 lb gain in maternal weight was associated with a 6 gm increase in birth weight. Birth weight was negatively correlated with a complicated obstetrical history and maternal cigarette smoking. Infants of heavy smokers weighed an average of 156 gm less than infants of nonsmokers. One-minute Apgar scores were significantly lower for infants of mothers who consumed alcohol and those who had had a complicated obstetrical history. Initial maternal weight and the husband's occupation positively influenced gestational age. There was no relationship between the incidence of maternal or infant complications and independent variables.


Full-Thickness Rotator Cuff Tears Can Be Safely Treated With a Resorbable Bioinductive Bovine Collagen Implant: One-Year Results of a Prospective, Multicenter Registry.
The purpose of this study was to prospectively collect safety and efficacy data in a large group of patients undergoing arthroscopic repair of full-thickness rotator cuff tears augmented with a resorbable bioinductive bovine collagen implant designed to promote healing. Seventeen centers across the United States enrolled patients in an institutional review board-approved registry to collect outcomes data on the implant. Patients undergoing surgical management of full-thickness rotator cuff tears augmented with the implant were enrolled. Inclusion criteria were age of ≥21 years, willingness to participate and the ability to read and speak English. Exclusion criteria included hypersensitivity to bovine-derived products. Patients were assessed before and after surgery at up to 1 year with outcomes including the single-assessment numeric evaluation (SANE), Veterans RAND 12-Item (VR-12) mental components and physical components (VR-12 PCS), American Shoulder and Elbow Surgeons (ASES), and Western Ontario Rotator Cuff (WORC) outcome measures. Ad hoc analyses were performed to compare these outcomes at all time points depending on tear size (small/medium vs large/massive). Serious complications were collected. Of 210 patients enrolled, 192 had 1-year follow-up data available. The patients experienced statistically significant improvement between baseline and 1 year for mean SANE, VR-12 PCS, ASES, and WORC scores (40.0-82.0, 33.5-47.3, 46.2-87.8, and 36.2-81.0, respectively; P < .001 for all results). Ad-hoc analysis demonstrated that similar results were obtained at 1 year regardless of tear size. Twenty patients (10.4%) experienced serious complications (10.4%), including revision surgery (n = 18), proximal humerus fracture/partial subscapularis tear resulting from multiple falls (n = 1), and adhesive capsulitis (n = 1). The safety and efficacy of a bioinductive implant in the surgical management of full-thickness rotator cuff tears at 1 year was shown in this study. Implant efficacy appears to be comparable regardless of the underlying tear size. Level IV, therapeutic case series.


Emerging role of metabolic reprogramming in tumor immune evasion and immunotherapy.
Mounting evidence has revealed that the therapeutic efficacy of immunotherapies is restricted to a small portion of cancer patients. A deeper understanding of how metabolic reprogramming in the tumor microenvironment (TME) regulates immunity remains a major challenge to tumor eradication. It has been suggested that metabolic reprogramming in the TME may affect metabolism in immune cells and subsequently suppress immune function. Tumor cells compete with infiltrating immune cells for nutrients and metabolites. Notably, the immunosuppressive TME is characterized by catabolic and anabolic processes that are critical for immune cell function, and elevated inhibitory signals may favor cancer immune evasion. The major energy sources that supply different immune cell subtypes also undergo reprogramming. We herein summarize the metabolic remodeling in tumor cells and different immune cell subtypes and the latest advances underlying the use of metabolic checkpoints in antitumor immunotherapies. In this context, targeting both tumor and immune cell metabolic reprogramming may enhance therapeutic efficacy.


Gender differences in lipoprotein lipase activity after acute exercise.
To determine whether gender differences exist in lipoprotein lipase (LPL) activity in response to exercise and/or insulin. Exercise and insulin are known modulators of LPL activity in men, but this is less clear in women. LPL activity may predict propensity for obesity; therefore, understanding its modulators is of considerable importance. Gender differences in skeletal muscle and adipose tissue LPL activity were determined after a single bout of exercise followed by a hyperinsulinemic/euglycemic clamp and compared with an identical rest day in healthy lean men (n = 10) and women (n = 10). Muscle and adipose tissue biopsies were obtained pre- (post-exercise vs. rest) and post-clamp. Basal levels of muscle and adipose tissue LPL activity were not different between men and women. There was, however, a significant gender by day interaction for muscle LPL activity (p = 0.023) and adipose tissue LPL activity (p = 0.013). In muscle, this was because of a significant increase in LPL activity on the exercise vs. rest day in men (p < 0.001) but not women. Adipose tissue LPL activity also increased significantly in men on the exercise day relative to rest day (p = 0.04) but decreased in women (p = 0.10). The hyperinulinemic/euglycemic clamp had no independent effect on tissue LPL activity, in either gender, after rest or exercise. In the 3 to 4 hours after exercise, muscle and adipose tissue LPL activity increased significantly in men, whereas LPL activity remained unchanged in women.


Time to pregnancy and semen parameters: a cross-sectional study among fertile couples from four European cities.
In fertile populations, little is known about the association between semen parameters and time to pregnancy (TTP). Pregnant women from Copenhagen, Edinburgh, Paris and Turku who conceived without medical intervention were asked for their TTP (942 couples), and their partners provided a semen sample. The proportion of morphologically normal sperm and the multiple anomalies index (MAI, ratio of the total number of anomalies to the number of abnormal sperm) were centrally estimated. We estimated rate ratios for the occurrence of a pregnancy by a discrete survival model, adjusted for sexual activity and female factors affecting fecundity. Increasing sperm concentration influenced TTP up to 55 x 10(6)/ml. The proportion of morphologically normal sperm influenced TTP up to 39% according to David's criteria, and this association held among the subjects with a sperm concentration >55 x 10(6)/ml. For strict criteria, the threshold value was 19% normal sperm. An increase of 0.5 in MAI was associated with an adjusted rate ratio for the occurrence of a pregnancy of 0.68 (95% confidence interval: 0.54-0.85). These results highlight the importance of sperm morphology parameters and indicate that the effect of proportion of normal sperm on TTP may be independent of sperm concentration.


Periodic acid Schiff--p phenylenediamine staining of glycogen in chondrocytes. A new combination which improves both cellular detail and glycogen identification.
This study reports a method whereby glycogen is identified in the chondrocytes of the secondary center of ossification prior to mineralization. The use of new fuchsin rather than basic fuchsin on one micron Spurr sections of femoral head cartilage fixed with potassium ferrocyanide-reduced osmium produced excellent identification of glycogen and when followed by p phenylenediamine, intensified cellular detail.


The rat urinary bladder: vasoactivity and macromolecular leakage in a new model.
An in vivo model of the rat urinary bladder microcirculation has been developed and microcirculatory responses to agents which produce vasoconstriction, vasodilation, and macromolecular leakage have been characterized. The urinary bladder of anesthetized female Sprague-Dawley rats is exteriorized and positioned in a tissue bath with a single stay suture which does not penetrate the lumen of the bladder. All blood vessels and nerves from the animal remain intact. The tissue bath is filled with Krebs solution which is monitored and maintained at a temperature of 36 +/- 0.5 degrees and a pH of 7.4 +/- 0.5. In vivo television microscopy is used to monitor vascular diameter and flow changes and isothiocyanate-tagged bovine serum albumin fluorescence is used as an index of macromolecular leakage. Norepinephrine (10(-6) M) caused a statistically significant decrease in vascular diameters of both arterioles and venules while sodium nitroprusside (10(-7) M) significantly increased arteriolar and venular diameters, histamine (10(-4) M) caused no change in venular diameters but did induce a significant macromolecular leak from those vessels. Compound 48/80 (1 and 10 micrograms/ml) induced a significant dose-dependent macromolecular leakage from venules. However, only with the 10 micrograms/ml dose was there visually detectable mast cell degranulation. It is concluded that this rat urinary bladder model provides a stable, reproducible model of a smooth muscle microcirculatory bed in a controlled environment, which responds similarly to other microcirculations.


Plant and Animal Innate Immunity Complexes: Fighting Different Enemies with Similar Weapons.
Both animals and plants express intracellular innate immunity receptors known as NLR (NOD-like receptors or nucleotide-binding domain and leucine-rich repeat receptors, respectively). For various mammalian systems, the specific formation of macromolecular structures, such as inflammasomes by activated NLR receptors, has been extensively reported. However, for plant organisms, the formation of such structures was an open scientific question for many years. This year, the first plant 'resistosome' structure was reported, revealing significant structural similarities to mammalian apoptosome and inflammasome structures. In this review, we summarize the key components comprising the mammalian apoptosome/inflammasome structures and the newly discovered plant resistosome, highlighting their commonalities and differences.


Account for high flow rate-low detrusor pressure voids in female: Contribution of VBN model.
Nomograms using the VBN model of women micturition allowed evaluating detrusor contractility (k) and urethral obstruction (U) from pressure-flow (PFs) recordings. While the model worked for most of the patients, an intriguing result, negative U value was observed for patients with high flow-low detrusor pressure (Qp voids). To explain that condition, our hypothesis was a weak urethral resistance to dilatation or increased expansibility (URD). The area offered to the fluid at each point of the urethra (its dilatation) is a function of the time depending difference between inside and outside pressures. In the VBN model, this function is sigmoid-like, the same for all women. For Qp voids, Qrecorded was more higher than it would be with the recorded pressure (VBN analysis). So, modeling allowed computing abnormally increased urethral wall expansibility (URD) whose consequence would be an increased flow. Among 222 non-neurologic women referred for investigation of various lower urinary tract symptoms, 27 (mean age 66.3±11.4 y) had Qp void: Qmax=27±6mL/s; pdet.Qmax =7.5±4.7cm H2O. Mean URD value was .36±.67. Introduction of URD in a modeled analysis of urodynamic traces led to a good fitting between recorded and computed traces for the 27 Qp. Mathematical modeling of micturition allows proposing an explanation of the unexpected observations of Qp voids. They would be due to abnormal urethral wall elasticity. Despite major challenges measurement of this elasticity would be the next step. 3.


Prolonged-release nicotinic acid for the management of dyslipidemia: an update including results from the NAUTILUS study.
Low HDL-cholesterol (<1.02 mmol/L [40 mg/dL] in men or <1.29 mmol/L [50 mg/dL] in women) occurs in about one-third of European patients with dyslipidemia and is an independent cardiovascular risk factor. Simultaneous correction of low HDL-cholesterol and high total-cholesterol and LDL-cholesterol may provide reductions in cardiovascular morbidity and mortality beyond those possible with statins alone. Nicotinic acid (niacin in the US) is the most effective means of increasing HDL-cholesterol available and has been shown to reduce cardiovascular event rates significantly. Niaspan (prolonged-release nicotinic acid) provides a convenient, once-daily means of administering nicotinic acid. Clinical studies with Niaspan have demonstrated marked, long-term increases in HDL-cholesterol with additional useful benefits on triglycerides, LDL-cholesterol, and lipid sub-profiles. The NAUTILUS study demonstrated the beneficial efficacy and tolerability profiles of Niaspan in a usual-care setting. The most common side-effect of Niaspan is flushing, which infrequently causes treatment discontinuation and which usually subsides over continued treatment. The ARBITER 2 and ARBITER 3 studies showed 1-2 years of treatment with Niaspan plus a statin induced regression of atherosclerosis in patients with coronary artery disease. The effect of Niaspan-statin treatment, relative to a statin alone, on clinical cardiovascular outcomes is currently under evaluation. Niaspan represents a practical means of correcting low HDL-cholesterol, an independent risk factor for adverse cardiovascular outcomes.


Application of the BrdU/thymidine method to flow cytogenetics: differential quenching/enhancement of Hoechst 33258 fluorescence of late-replicating chromosomes.
Discrimination between many types of isolated mammalian chromosomes can be accomplished by dual-beam flow cytometry following DNA staining with Hoechst 33258 (HO) and Chromomycin A3 (CA3). In this report, we show that the bivariate discrimination of selected late-replicating Chinese hamster M3-1 chromosomes can be improved by appropriate treatment of the cells with 5-bromo-2'-deoxyuridine (BrdU) prior to chromosome isolation and staining. Two labeling schemes are reported. In one scheme the chromosomes are collected from cells labeled with BrdU only during late S phase. The Hoechst fluorescence of the 10, 11, M2, and Y chromosomes is substantially quenched by the incorporated BrdU, thus improving their discrimination. In the other scheme, chromosomes are collected from cells labeled with thymidine (dT) during late S phase following 20 h of growth in BrdU-containing medium. The Hoechst fluorescence of the 10, 11, M2, and Y chromosomes is quenched less than the other chromosomes, again improving their discrimination. Y chromosomes from chromosome suspensions of untreated controls, of cells labeled with BrdU during late S phase, and of cells labeled with dT during late S phase following 20 h growth in BrdU were separated by dual-parameter sorting. While the purity of the sorted Y chromosome was 15% in untreated controls, it was 70-75% using the BrdU/dT labeling protocols.


Mechanism of oligomerization of Escherichia coli carbamoyl phosphate synthetase and modulation by the allosteric effectors. A site-directed mutagenesis study.
We use site-directed mutagenesis to clarify the role of effector-mediated oligomerization changes on the modulation of the activity of Escherichia coli carbamoyl phosphate synthetase (CPS) by its allosteric activator ornithine and its inhibitor UMP. The regulatory domain mutations H975L, L990A and N992A abolished, and N987V decreased CPS oligomerization. The oligomerization domain mutation L421E prevented tetramer but not dimer formation. None of the mutations had drastic effects on enzyme activity or changed the sensitivity or apparent affinity of CPS for ornithine and UMP. Our findings exclude the involvement of oligomerization changes in the control of CPS activity, and show that CPS dimers are formed by the interactions across regulatory domains, and tetramers by the interactions of two dimers across the oligomerization domains. A mechanism for effector-mediated changes of the oligomerization state is proposed.


[Benign mimickers of the prostate cancer. Diagnostic challenges].
The diagnosis of prostate cancer (PCa), especially limited adenocarcinoma on needle biopsy, is often challenging. Before making diagnosis of PCa, it is prudent for the pathologist to consider different benign patterns that may lead to a false positive interpretation. Histoanatomic structures such as seminal vesicles, Cowper's glands and paraganglia along with hyperplasia, atrophy with its different patterns and adenosis may generate difficulties in differential diagnosis. Furthermore, inflammatory processes and post-treatment changes may cause problems. The above entities can in some instances simulate low-grade and less commonly high grade PCa. Knowledge of these patterns and application of appropriate immunohistochemistry will lead the pathologist to a correct diagnosis.


Osimertinib Regresses an EGFR-Mutant Cisplatinum- Resistant Lung Adenocarcinoma Growing in the Brain in Nude Mice.
The goal of the present study was to determine the efficacy of osimertinib (AZD9291), a third-generation epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor for the treatment of aggressive EGFR-mutant non-small cell lung cancer (NSCLC), compared to cisplatinum (CDDP) + pemetrexed (PEM). The NSCLC cell line PC-9 expressing green fluorescence protein (PC-9-GFP) was implanted in the brain of nude mice and was treated with CDDP + PEM or AZD9291. Tumors were observed by non-invasive fluorescence imaging. AZD9291 treatment caused tumor regression in contrast to CDDP + PEM which had only a slight inhibitory effect. These results suggest that AZD9291 is a promising clinical option for NSCLC patients with brain metastasis.


Proteins associated with activity of Fc receptors on isolated human placental syncytiotrophoblast microvillous plasma membranes.
To identify Fc receptors from human placental microvilli, proteins that were liberated by detergents from human placental synctiotrophoblast microvillous membranes (StMPM) were characterized by their abilities to bind human IgG in immune complexes with sheep or goat anti-human IgG and to monomeric rabbit anti-dinitrophenol (DNP) IgG bound to DNP-lysine Sepharose. Three placental IgG-binding proteins coprecipitated with immune complexes (Mr = 68,000, 52,000-56,000, 40,000) and were designated pIBP68, pIBP56 and pIBP40, respectively. Of the three proteins only pIBP56 bound to immobilized monomeric rabbit IgG. It was isolated from detergent lysates of StMPM and LDS/phenol glycoprotein extracts of placental plasma membranes suggesting that pIBP56 was a glycoprotein FcR previously reported (Mikulska et al, 1982). The binding specificities of pIBP56 and pIBP40 appeared to be detergent dependent. Photoaffinity crosslinking of StMPM surface proteins in situ to monomeric rabbit derivatized with N-succinimidyl(4-azidophenyl)-I, 3-dithiopropionate identified IgG-binding proteins identical in size to pIBP56 and pIBP40. Crosslinking further suggested that monomeric IgG covalently bound to a complex of StMPM proteins with a total size of 110,000-120,000 Mr. The findings suggest that pIBP68, pIBP56 and pIBP68 are responsible for IgG binding activity of placental StMPM.


The A domain of clonal complex 1-type fibronectin binding protein B promotes adherence and biofilm formation in Staphylococcus aureus.
Adhesive interactions between Staphylococcus aureus and the host rely on cell-wall-anchored proteins such as fibronectin-binding protein B (FnBPB). Recently we showed that the FnBPB protein expressed by clonal complex (CC) 1 isolates of S. aureus mediates bacterial adhesion to corneodesmosin. The proposed ligand-binding region of CC1-type FnBPB shares just 60 % amino acid identity with the archetypal FnBPB protein from CC8. Here we investigated ligand binding and biofilm formation by CC1-type FnBPB. We found that the A domain of FnBPB binds to fibrinogen and corneodesmosin and identified residues within the hydrophobic ligand trench in the A domain that are essential for the binding of CC1-type FnBPB to ligands and during biofilm formation. We further investigated the interplay between different ligands and the influence of ligand binding on biofilm formation. Overall, our study provides new insights into the requirements for CC1-type FnBPB-mediated adhesion to host proteins and FnBPB-mediated biofilm formation in S. aureus.


Advanced fibrosis associated with non-alcoholic steatohepatitis (NASH) in Spain: results of a Delphi study.
To describe in detail the epidemiology, diagnosis, clinical management, treatment options, impact on quality of life and unmet needs of patients with advanced liver fibrosis (F3-F4) associated with non-alcoholic steatohepatitis (NASH) in Spain. Delphi study of two rounds of consultation rounds with 41 expert hepatologists from 16 autonomous communities to collect their experience in clinical practice. The estimated prevalence of adult patients diagnosed with F3-F4 fibrosis associated with NASH in Spain is 0.019% (95% confidence interval [CI]: 0.019-0.020%). Approximately 7,588 adults with this condition are currently diagnosed and managed in the Digestive System Services of Spanish hospitals, and around 1,881 new patients are diagnosed each year. Management is multidisciplinary and includes the specialties of Digestive System, Endocrinology and Internal Medicine, considering the frequently associated metabolic comorbidities (obesity, type 2 diabetes mellitus or dysmetabolic iron overload). Despite a clear impact on quality of life, this it is not routinely evaluated in clinical practice. The most widely used non-invasive diagnostic techniques are transitional elastography and liver fibrosis index 4 (FIB-4). The absence of effective and safe treatments appears as the main unmet need for the management of these patients. This study provides a representation of the current situation of patients diagnosed with F3-F4 fibrosis associated with NASH in Spain, increasing the evidence available and contributing to informed decision-making by professionals and the health system.


[Possibilities of surgical treatment of gastric cancer recurrences depending on the type of previous resection].
The study deals with the end results of 375 surgeries performed for recurrent cancer of the stomach. Radical surgery was carried out in 168 cases (extirpation of the remaining part of stomach--140; repeated resection of stomach--12; extirpation of esophagoenterostomy--16). Stomach resection after Billroth-II with anterior long-loop colostomy created the best conditions for radical extirpation of the remaining stomach, while Billroth-I resection--the worst. The end results were worse when tumor relapsed earlier than three years after primary resection of the stomach and on sites in gastroenterostomy area, as compared to relapse at later stage and outside anastomosis area.


A systematic review of economic evaluations of pharmacist services.
Challenges to the provision of health care are occurring internationally and are expected to increase in the future, further increasing health spending. As pharmacist roles are evolving and expanding internationally to provide individualised pharmaceutical care it is important to assess the cost-effectiveness of these services. To systematically synthesise the international literature regarding published economic evaluations of pharmacy services to assess their cost-effectiveness and clinical outcomes. A systematic review of economic evaluations of pharmacy services was conducted in MEDLINE, EMBASE, PubMed, Scopus, Web of Science, CINAHL, IPA and online journals with search functions likely to publish economic evaluations of pharmacy services. Data were extracted regarding the interventions, the time horizon, the outcomes and the incremental cost-effectiveness ratio. Studies' quality of reporting was assessed using the Consolidated Health Economic Evaluation Reporting Standard (CHEERS) statement. Seventy-five studies were included in the systematic review, including 67 cost-effectiveness analyses, 6 cost-benefit analyses and 2 cost-consequence analyses. Of these, 57 were either dominant or cost-effective using a willingness-to-pay threshold of NZ$46 645 per QALY. A further 11 studies' cost-effectiveness were unable to be evaluated. Interventions considered to be most cost-effective included pharmacist medication reviews, pharmacist adherence strategies and pharmacist management of type 2 diabetes mellitus, hypertension and warfarin/INR monitoring. The quality of reporting of studies differed with no studies reporting all 28 items of the CHEERS statement. There is strong economic evidence to support investment in extended pharmacist services, particularly those focussed on long-term chronic health conditions.


Comparison of the effects of two intrathecal anaesthetic techniques for transurethral prostatectomy on haemodynamic and pulmonary function.
Transurethral prostatectomy is routinely performed under spinal anaesthesia. This technique can cause hypotension, which is particularly undesirable in the elderly. The objective was to compare spinal anaesthesia for transurethral prostatectomy using hyperbaric bupivacaine 15 mg (control group) and hyperbaric bupivacaine 10 mg (limiting spread by maintaining the upright position for 15 min) and fentanyl 25 microg (fentanyl group) in terms of haemodynamic and pulmonary function. Thirty ASA I-III patients were randomly selected and underwent spinal anaesthesia with either hyperbaric bupivacaine 15 mg (immediately positioned supine) or hyperbaric bupivacaine 10 mg (upright for 15 min) and fentanyl 25 microg. The greatest changes in mean arterial pressure (P = 0.9), ephedrine requirements (P = 0.8) and mean maximum change in forced vital capacity (P = 0.5) were similar in both groups. The addition of fentanyl 25 microg to bupivacaine 10 mg and limiting the spread of the block does not improve either haemodynamic or pulmonary function compared with bupivacaine 15 mg in patients undergoing transurethral prostatectomy.


Is Endomyocardial Biopsy a Safe and Useful Procedure in Children with Suspected Cardiomyopathy?
Endomyocardial biopsy (EMB) is a common procedure used to aid in the diagnosis, prognosis and treatment of suspected pediatric cardiomyopathy. In suspected cardiomyopathy, no multicenter experience has previously reported on the safety and utility of EMBs. Retrospectively, adverse event (AE) and patient and procedural characteristics were obtained at seven institutions participating in the Congenital Cardiac Catheterization Outcomes Project for both a cardiomyopathy (n = 158) and a post-transplant surveillance (n = 2665) cohort. Descriptive information regarding biopsy indication, pathology and clinical management based on EMB findings were retrospectively obtained. High-severity AEs were more common in the cardiomyopathy cohort when compared to the post-transplant surveillance cohort. The cardiomyopathy cohort was younger, more hemodynamically vulnerable and required more cardiorespiratory support during the procedure. The eight high-severity AEs in the cardiomyopathy group included one myocardial perforation, two ECMO cannulations and three deaths following the EMB. Factors associated with high-severity AEs included performing another catheter-based intervention during the EMB and longer fluoroscopy time. Notably, an increased number of biopsy attempts did not increase the risk of an AE. Suspected myocarditis was the most common indication. Diagnostic EMB pathology and thus alteration to clinical management based on pathology occurred more frequently in patients with suspected myocarditis. In conclusion, there is an increased incidence of high-severity AEs in patients undergoing EMB for suspected cardiomyopathy. EMB may be more clinically useful in the management of suspected myocarditis. The increased risk of high-severity AEs when additional interventions are performed highlights the hemodynamic vulnerability in patients with suspected cardiomyopathy.


The Sheppe and Hain study revisited: professional students and their knowledge and attitudes about human sexuality.
The professions of law and medicine exert a profound influence on sexual attitudes and behaviour in our society. Previous reported studies of legal and medical students found no differences in sexual knowledge for entering first-year students. However, in this study, a sample of 160 medical and seventy-nine law students revealed significant differences in knowledge and attitudes about sexuality. The data indicate that a gender-linked double standard of acceptable behaviour is not present. Results further reinforce the necessity of medical sex education.


Organelle positioning and cell polarity.
In spite of conspicuous differences in their polarized architecture, swimming unicellular eukaryotes and migrating cells from metazoa display a conserved hierarchical interlocking of the main cellular compartments, in which the microtubule network has a dominant role. A microtubule array can organize the distribution of endomembranes owing to a cell-wide and polarized extension around a unique nucleus-associated structure. The nucleus-associated structure in animal cells contains a highly conserved organelle, the centriole or basal body. This organelle has a defined polarity that can be transmitted to the cell. Its conservative mode of duplication seems to be a core mechanism for the transmission of polarities through cell division.


Dose-dependent accumulation of glucose in the intestinal wall and lumen induced by metformin as revealed by 18 F-labelled fluorodeoxyglucose positron emission tomography-MRI.
To investigate the relationships between various clinical variables and the metformin-induced accumulation of fluorodeoxyglucose (FDG) in the intestine, with distinction between the intestinal wall and lumen, in individuals with type 2 diabetes who were receiving metformin treatment and underwent 18 F-labelled FDG ([18 F]FDG) positron emission tomography (PET)-MRI. We evaluated intestinal accumulation of [18 F]FDG with both subjective (a five-point visual scale determined by two experienced radiologists) and objective analyses (measurement of the maximum standardized uptake value [SUVmax ]) in 26 individuals with type 2 diabetes who were receiving metformin and underwent [18 F]FDG PET-MRI. [18 F]FDG accumulation within the intestinal wall was discriminated from that in the lumen on the basis of SUVmax . SUVmax for the large intestine was correlated with blood glucose level (BG) and metformin dose, but not with age, body mass index, HbA1c level or estimated glomerular filtration rate (eGFR). SUVmax for the small intestine was not correlated with any of these variables. Visual scale analysis yielded essentially similar results. Metformin dose and eGFR were correlated with SUVmax for the wall and lumen of the large intestine, whereas BG was correlated with that for the wall. Multivariable analysis identified metformin dose as an explanatory factor for SUVmax in the wall and lumen of the large intestine after adjustment for potential confounders including BG and eGFR. Metformin dose is an independent determinant of [18 F]FDG accumulation in the wall and lumen of the large intestine in individuals treated with this drug.


Selectively attending to natural scenes after alcohol consumption: an ERP analysis.
Alcohol effects on cognitive, emotional and behavioral processes have been linked to an impairment of attention. Because attention operates at the level of specific cognitive subsystems, recent studies demonstrated alcohol effects in specific post-perceptual processes such as response selection and working memory. Measuring event-related potentials, the present study focused on perceptual processes by utilizing a categorization task where participants had to decide whether briefly presented images contained an animal or not. Findings demonstrate an early differential ERP activity for target compared to non-target images, which was reduced after alcohol intoxication. Thus, alcohol intoxication had deleterious effects at the perceptual level of processing considered to reflect the interaction of top-down (category-related) and bottom-up (stimulus-driven) processes. In addition, post-perceptual processes were also impaired by alcohol intoxication.


Synthesis and ex vivo evaluation of carbon-11 labelled N-(4-methoxybenzyl)-N'-(5-nitro-1,3-thiazol-2-yl)urea ([11C]AR-A014418): a radiolabelled glycogen synthase kinase-3beta specific inhibitor for PET studies.
N-(4-Methoxybenzyl)-N'-(5-nitro-1,3-thiazol-2-yl)urea (AR-A014418), a highly selective inhibitor of glycogen synthase kinase-3beta (GSK-3beta), was radiolabelled with carbon-11 (half-life=20.4min) for cerebral positron emission tomography (PET) studies. Reaction of desmethyl AR-A014418 with [(11)C]CH(3)I produced [(11)C]AR-A014418 in 17% decay-corrected radiochemical yield, based on [(11)C]CO(2), with 3230mCi/micromol specific activity after a 30min synthesis time. The desmethyl precursor of AR-A014418 was synthesized in 23% yield by a novel one-pot reaction of 2-amino-5-nitrothiazole with in situ generated TMS-protected 4-hydroxybenzylisocyanate, following deprotection with acid. Ex vivo biodistribution studies were conducted after [(11)C]AR-A014418 was administered via tail vein injection into Sprague-Dawley rats. Very low levels of radioactivity were found in all brain regions (0.08% injected dose/gram of tissue) at 5 and 30min post-injection, uncorrected for vascular compartment. Considering the extremely poor brain penetration of [(11)C]AR-A014418 this compound cannot be used to study GSK-3beta in cerebral PET studies. Furthermore, the specific pharmacological mechanism(s) of antidepressant-like activity attributed to AR-A014418 should be investigated.


Intracerebroventricular injection of murine leptin enhances the postprandial metabolic rate in the rat.
Energy balance is achieved by means of a concomitant control of both food intake and energy expenditure. Leptin, synthesized in the adipose tissue, acts on brain structures and lowers body weight by inhibiting food intake and in parallel by enhancing energy expenditure i.e. metabolism or one of its components. Recording distinctly these components allowed us to assess the effect of an acute intracerebroventricular injection of leptin on both feeding pattern and background metabolism (i.e. energy expenditure free from the part of locomotor activity), respiratory quotient, feeding-related metabolism and locomotor activity-related metabolism. Leptin injection to Sprague-Dawley male rats induced an inhibition of feeding that began 90 min after the treatment and lasted 1 h before to return to the control feeding pattern level. Considering this late behavioral effect, it appeared that leptin may act during the postprandial period so that we recorded the different metabolic parameters following a 3 g calibrated meal itself preceded by leptin vs. artificial cerebrospinal fluid injection. Postprandial respiratory quotient was rapidly lowered in leptin-treated animals and subsequent background metabolism increased for 6 h. Thus it appeared that leptin increased the duration of the postprandial metabolic rate via the recruitment of endogenous fat stores. Enhancement in the thermic effect of food may be the reason for feeding behavior inhibition to be delayed.


nus-tool: A unified program for generating and analyzing sample schedules for nonuniformly sampled NMR experiments.
Increases in digital resolution achieved by high-field NMR require increases in spectral width. Additionally, the ability to resolve two overlapping peaks requires a sufficiently long acquisition time. These constraints combine, so that achieving high resolution spectra on high-field magnets requires long experiment times when employing uniform sampling and Fourier Transform processing. These limitations may be addressed by using nonuniform sampling (NUS), but the complexity of the parameter space across the variety of available NUS schemes greatly hinders the establishment of optimal approaches and best practices. We address these challenges with nus-tool, which is a software package for generating and analyzing NUS schedules. The nus-tool software internally implements random sampling and exponentially biased sampling. Through pre-configured plug-ins, it also provides access to quantile sampling and Poisson gap sampling. The software computes the relative sensitivity, mean evolution time, point spread function, and peak-to-sidelobe ratio; all of which can be determined for a candidate sample schedule prior to running an experiment to verify expected sensitivity, resolution, and artifact suppression. The nus-tool package is freely available on the NMRbox platform through an interactive GUI and via the command line, which is especially useful for scripted workflows that investigate the effectiveness of various NUS schemes.


A BODIPY-based turn-on fluorescent probe for the selective detection of hydrogen sulfide in solution and in cells.
A new turn-on fluorescent probe, incorporating 7-nitrobenzoxadiazole (NBD) ether group into a BODIPY molecule, was synthesized and studied for the detection of H2S and biothiols in aqueous solution and in living cells. The design was based on thiol-induced thiolysis of the NBD ether bond and followed by the cleavage and the release of free meso-(4-hydroxybenzyl)-substituted BODIPY fluorophore. The BODIPY-based probe displayed highly sensitive and selective fluorescence enhancement respond to H2S over competing biothiols such as cysteine (Cys) and glutathione (GSH), mainly due to the weak reactivity of biothiols toward the probe. There was a good linearity between the fluorescence intensity and the concentrations of H2S in the range of 1-200 µM with a detection limit of 2.6 µM. The proposed reaction mechanism was confirmed by mass spectrometry and optical spectroscopy, and the mechanism of turn-on fluorescent response was further determinated by the density functional theory (DFT) calculations using Gaussian 03 program. Moreover, the probe was successfully applied for the fluorescence imaging of H2S in HeLa cells under physiological conditions (pH 7.4).


Double scattering and pencil beam scanning Monte Carlo workflows for proton therapy retrospective studies on radiation-induced toxicities.
Monte Carlo (MC) simulations can be used to accurately simulate dose and linear energy transfers (LET) distributions, thereby allowing for the calculation of the relative biological effectiveness (RBE) of protons. We present hereby the validation and implementation of a workflow for the Monte Carlo modelling of the double scattered and pencil beam scanning proton beamlines at our institution. The TOPAS/Geant4 MC model of the clinical nozzle has been comprehensively validated against measurements. The validation also included a comparison between simulated clinical treatment plans for four representative patients and the clinical treatment planning system (TPS). Moreover, an in-house tool implemented in Python was tested to assess the variable RBE-weighted dose in proton plans, which was illustrated for a patient case with a developing radiation-induced toxicity. The simulated range and modulation width closely matches the measurements. Gamma-indexes (3%/3mm 3D), which compare the TPS and MC computations, showed a passing rate superior to 98%. The calculated RBE-weighted dose presented a slight increase at the necrosis location, within the PTV margins. This indicates the need for reporting on the physical and biological effects of irradiation in high dose regions, especially at the healthy tissues and increased LET distributions location. The results demonstrate that the Monte Carlo method can be used to independently validate a TPS calculation, and to estimate LET distributions. The features of the in-house tool can be used to correlate LET and RBE-weighted dose distributions with the incidence of radiation-induced toxicities following proton therapy treatments.


Aging impairs recovery from stress-induced depression in male rats possibly by alteration of microRNA-101 expression and Rac1/RhoA pathway in the prefrontal cortex.
Along with altering brain responses to stress, aging may also impair recovery from depression symptoms. In the present study, we investigated depressive-like behaviors in young and aged rats and assayed the levels of microRNA-101 (miR-101), Rac1/RhoA, PSD-95, and GluR1 in the prefrontal cortex (PFC) after stress cessation and after a recovery period. Young (3 months old) and aged (22 months old) male Wistar rats were divided into six groups; Young control (YNG), young rats received chronic stress for four weeks (YNG + CS), young rats received chronic stress for four weeks followed by a 6-week recovery period (YNG + CS + REC), Aged control (AGED), aged rats received chronic stress for four weeks (AGED + CS), and aged rats received chronic stress for four weeks followed by a 6-week recovery period (AGED + CS + REC). Stress-induced depression, evaluated by the sucrose preference test (SPT) and forced swimming test (FST), was yet observed after the recovery period in aged but not in young rats, which were accompanied by unchanged levels of miR-101, Rac1/RhoA, GluR1, and PSD-95 in the PFC of aged rats. These data suggested that impaired synaptic plasticity of glutamatergic synapses via the miR-101/Rac1/RhoA pathway may contribute to the delayed behavioral recovery after stress exposure observed in aging animals.


Validation of 4D Monte Carlo dose calculations using a programmable deformable lung phantom.
To validate the accuracy of 4D Monte Carlo (4DMC) simulations to calculate dose deliveries to a deforming anatomy in the presence of realistic respiratory motion traces. A previously developed deformable lung phantom comprising an elastic tumor was modified to enable programming of arbitrary motion profiles. 4D simulations of the dose delivered to the phantom were compared with the measurements. The deformable lung phantom moving with irregular breathing patterns was irradiated using static and VMAT beam deliveries. Using the RADPOS 4D dosimetry system, point doses were measured inside and outside the tumor. Dose profiles were acquired using films along the motion path of the tumor (S-I). In addition to dose measurements, RADPOS was used to record the motion of the tumor during dose deliveries. Dose measurements were then compared against 4DMC simulations with EGSnrc/4DdefDOSXYZnrc using the recorded tumor motion. The agreements between dose profiles from measurements and simulations were determined to be within 2%/2 mm. Point dose agreements were within 2σ of experimental and/or positional/dose reading uncertainties. 4DMC simulations were shown to accurately predict the sensitivity of delivered dose to the starting phase of breathing motions. We have demonstrated that our 4DMC method, combined with RADPOS, can accurately simulate realistic dose deliveries to a deforming anatomy moving with realistic breathing traces. This 4DMC tool has the potential to be used as a quality assurance tool to verify treatments involving respiratory motion. Adaptive treatment delivery is another area that may benefit from the potential of this 4DMC tool.


Identification of Dutch children diagnosed with atopic diseases using prescription data: a validation study.
The aim of this study is to validate medication proxies for the identification of children diagnosed with atopic disorders that can be applied in various types of epidemiological research. Records of 7439 children, aged between 0 and 10 years, in the period 2001 until 2010, were retrieved from the Registration Network Groningen database, a general practitioners database in the north-eastern part of the Netherlands. The sensitivity and positive predictive value (PPV) of 22 medication proxies for the identification of children diagnosed with atopic disorders (asthma, atopic dermatitis, and allergic rhinitis) were computed using the registered diagnoses as gold standards. In addition, different capture periods (1 year, half year, and length of study period) for the detection of prescriptions were tested for all the medication proxies. The highest PPV (0.84, 95 % CI 0.81-0.87) in combination with a sufficient sensitivity value (0.54, 95 % CI 0.50-0.57) for the identification of children diagnosed with asthma was yielded for the medication proxy, ≥2 prescriptions for anti-asthma medication within 1 year, including 1 inhaled steroid. PPV and sensitivity were even higher in the age group 6-10 years. The proxies designed for the identification of children diagnosed with atopic dermatitis and allergic rhinitis yielded only high PPVs (≥0.75) in combination with low sensitivity values (≤0.22). Altering the capture period for the detection of prescriptions to half a year or the length of the study period only affected sensitivity values. Children diagnosed with asthma can be identified reliably with a range of medication proxies. The use of prescription data for the identification of children diagnosed with atopic dermatitis and allergic rhinitis is questionable.


Best Practices in Relational Skills Training for Medical Trainees and Providers: An Essential Element of Addressing Adverse Childhood Experiences and Promoting Resilience.
Medical providers' ability to form strong therapeutic alliances with patients is an essential clinical skill that is associated with a higher quality of care and improved provider well-being. However, comparatively few medical providers exhibit adequate relational skills, which serve to convey respect, communicate caring, and build trust between the medical provider and the patient. A growing number of medical training programs and continuing medical education programs have begun to incorporate relational skills training, but the results have been highly variable in terms of training methods and effect. To support administrators who are considering the implementation (or improvement) of relational skills training in their organization, we provide a set of best practices for relational skills training, in the basis of a review of the literature and on our experience as clinical educators, and show the application of these best practices through a case study. We conclude with a discussion of challenges for implementing a high-quality relational skills training program, policy-level solutions for these challenges, and recommendations for future research.


Of mice and chickens: Revisiting the RC time constant problem.
Avian hair cells depend on electrical resonance for frequency selectivity. The upper bound of the frequency range is limited by the RC time constant of hair cells because the sharpness of tuning requires that the resonance frequency must be lower than the RC roll-off frequency. In contrast, tuned mechanical vibration of the inner ear is the basis of frequency selectivity of the mammalian ear. This mechanical vibration is supported by outer hair cells (OHC) with their electromotility (or piezoelectricity), which is driven by the receptor potential. Thus, it is also subjected to the RC time constant problem. Association of OHCs with a system with mechanical resonance leads to piezoelectric resonance. This resonance can nullify the membrane capacitance and solves the RC time constant problem for OHCs. Therefore, avian and mammalian ears solve the same problem in the opposite way. This article is part of the Special Issue Outer hair cell Edited by Joseph Santos-Sacchi and Kumar Navaratnam.


Suppression tumorigenicity 2 (ST2) turbidimetric immunoassay compared to enzyme-linked immunosorbent assay in predicting survival in heart failure patients with reduced ejection fraction.
Suppressor of tumorigenicity 2 (ST2) is a powerful marker of prognosis and treatment response in heart failure (HF), however, it is an enzyme-linked immunosorbent assay (ELISA) which may be cumbersome and costly. A turbidimetric immunoassay (TIA) that can run on common chemistry analyzers could overcome this. We studied a novel TIA for ST2, comparing it to commercial ST2 (ELISA). Patients age ≥ 18 years meeting Framingham definition for HF were enrolled in a prospective registry (Oct 2007 - March 2015) at Henry Ford Hospital and donated blood samples. Participants with reduced ejection fraction (<50%) and available plasma samples were included and valid ST2 measurements were obtained on the same sample using both TIA and ELISA (N = 721). The primary endpoint was all cause death. Correlation between the methods was quantified. The association with survival was tested using unadjusted and adjusted (for MAGGIC score and NTproBNP) Cox models and comparing the Area Under the Curve (AUC). The inter-assay Spearman correlation coefficient was 0.77. Nonparametric regression showed no significant proportional difference (slope = 0.97) and a very small systematic difference (3.2 ng/mL). In univariate analyses, both TIA and ELISA ST2 were significant associates of survival with similar effect sizes (HR 4.46 and 3.50, respectively, both p < 0.001). In models adjusted for MAGGIC score, both ST2 remained significant in Cox models and incrementally improved AUC vs. MAGGIC alone (MAGGIC AUC = 0.757; TIA + MAGGIC AUC = 0.786, p = 0.025; ELISA + MAGGIC AUC = 0.793, p = 0.033). In models with both MAGGIC and NTproBNP included, both ST2 still remained significant but did not improve AUC. A novel TIA method for ST2 quantification correlates highly with ELISA and offers similarly powerful risk-stratification.


Association of glucocorticoid receptor gene polymorphisms with systemic lupus erythematosus in a Chinese population.
To identify the association of glucocorticoid receptor (GR) gene polymorphism with systemic lupus erythematosus (SLE). A case-control study was carried out, in which 400 Chinese patients with SLE and 400 normal people were enrolled. DNA was extracted using a genomic DNA extraction kit, and tagged single nucleotide polymorphisms (SNPs) were identified by Haploview (4.0) Project from the Chinese HapMap Project. Eighteen tagged SNPs of the GR gene were genotyped by the Multiplex SNaPshot technique. Two GR gene SNPs were associated with the pathogenesis of SLE: rs6865292 (dominant model: crude odds ratio [OR] = 1.526, 95% CI: 1.151-2.025, P = 0.003; adjusted OR = 1.525, 95% CI: 1.149-2.023, P = 0.004; PBH = 0.036) and rs9324921 (dominant model: crude OR = 1.556, 95% CI: 1.173-2.062, P = 0.002; adjusted OR = 1.553, 95% CI: 1.171-2.060, P = 0.002; PBH = 0.036). The haplotype analysis of GR gene SNPs manifested that the haplotype of 'CCGGG' (OR = 2.701, 95% CI: 1.348-5.410, P = 0.004; PBH = 0.036) was a risk factor for the development of SLE. A lower frequency of A-allele of SNP rs4607376 (P = 0.021; OR = 0.794, 95% CI: 0.652-0.966, PBH = 0.126), higher frequency of C-allele of SNP rs6865292 (P = 0.019, OR = 1.317, PBH = 0.126) and A-allele of SNP rs9324921 (P = 0.019, OR = 1.317, PBH = 0.126) may be risk factors for developing SLE. The rs7719514 (recessive model: crude P = 0.044; adjusted P = 0.044, PBH = 0.264), rs7701443 (recessive model: crude P = 0.044, adjusted P = 0.045; PBH = 0.264), rs4607376 (recessive model: crude P = 0.027; adjusted P = 0.026; PBH = 0.264) and haplotype 'CAGCG' (P = 0.044; PBH = 0.198) showed marginal association with the pathogenesis of SLE. In the case group, there were no significant differences between non-lupus nephritis and lupus nephritis. Further, we found that the SNP rs12054797 (F = 3.228, P = 0.041, PBH = 0.342), rs2963156 (F = 3.163, P = 0.043, PBH = 0.342) might be marginally associated with disease activity. The study indicates that GR genetic polymorphisms may play a major role in the pathogenesis and development of SLE.


Reasons for not performing refractive surgery.
To analyze the reasons that laser in situ keratomileusis (LASIK) or photorefractive keratectomy (PRK) were not performed in patients who requested surgical correction of their refractive errors. Minamiaoyama Eye Clinic, Tokyo, Japan. This retrospective review comprised 2784 consecutive patients who visited our clinic between June 1997 and August 2000. The reasons they did not receive refractive surgery (PRK or LASIK) were analyzed. Among the 2784 patients who requested surgery for refractive errors, 2079 patients (74.7%) had PRK or LASIK and 705 patients (25.3%) did not. The most common reasons for not treating patients surgically were myopia greater than -12.0 diopters and/or high astigmatism (20.7%), insufficient corneal thickness (8.2%), keratoconus (6.4%), cataract (5.7%), and hyperopia and/or hyperopic astigmatism (4.1%). Patients who request surgery have a variety of problems. Attention must be given to these individual problems, and the indications must be considered.


Characterization and management of mandibular fractures: lessons learned from Iraq and Afghanistan.
Fractures of the mandible are among the most common facial injuries. Invasiveness of treatment should be determined by the extent of injury: degree of displacement, number of fractures, the patient's health status, and concomitant injuries. Complex, comminuted, and avulsive injuries frequently seen in combat will require coordination with multiple specialties to provide the best treatment. Stabilization treatment with arch bars or external fixators and splints is often desirable when fractures are highly comminuted or the soft tissue envelope is compromised by tissue loss or burns. In severe injuries, many times reconstruction will take several surgeries. Debridement of necrotic tissue and devascularized bone and skin grafting often are necessary before reconstruction. Microvascular or myocutaneous flaps should be considered with significant tissue loss and osteocutaneous flaps when large continuity defects are present. Most mandible fractures are repaired in a single operation. Those caused by explosives and high-velocity projectiles are more complex. Research should continue to focus on improving outcomes for these patients. Advances in tissue engineering, bone regeneration, and composite tissue allografting will have to continue if we hope to restore facial form and function for our combat wounded.


Policy changes for preventing and recognizing overwork-related cardiovascular diseases in Taiwan: An overview.
Overwork-related cardiovascular and cerebrovascular disease (CVD) has a large impact on workers' rights and labor standards across East Asian countries. This article describes the background and impact of policies regarding overwork-related CVD in the past decades in Taiwan. We reviewed government policies, guidelines, literature, and news addressing the problems and impact of policy changes to prevent and recognize overwork-related CVD since the 1990s, and collected data on overwork-related CVD cases in Taiwan from 2006 to 2017. In 2017, overwork-related CVD accounted for 13% of all cases of occupational diseases, but 79% of all deaths due to occupational diseases. Guidelines for recognizing overwork-related CVD cases were established in 1991; however, under-recognition exists in Taiwan due to poor exposure data on working hours and psychological factors and because most medical expenses are covered by the national health insurance system. Amendments on the guidelines, and stricter policies on overwork prevention were enforced following calls from labor unions, nongovernmental organizations, and legislators, but health disparities were introduced when certain industries were exempted from restrictions. Long working hours and other work characteristics increase the risk of overwork-related CVD. By reviewing the changes in policy for preventing overwork and recognizing overwork-related disease, we identified the need for clearly defined guidance on evaluating overwork-related CVD, with specific criteria for working hours and other risk factors regarding work characteristics. National policies that lead to better working conditions and prevent overwork-related diseases must be developed.


Meta-analysis of MspI derived variants of growth hormone gene associated with milk yield in dairy cattle.
The present work aimed to obtain common effect sizes for the gene frequency and association of MspI derived variants of growth hormone (GH) gene with milk yield in dairy cows. We performed a systematic review and meta-analysis of 35 published studies identified in literature search from 2000 to 2020 (n = 4164). These studies were specific to fragment size (329) for genotypes viz., CC (224, 105 bp), CD (329, 224, 105 bp) and DD (329 bp). Pooled standard mean differences (SMDs) as effect sizes between allele pairs were derived using different genetic models. The heterogeneity between effects sizes across studies was estimated using I2 Index (%). The common effect size for gene frequency of allele C (224, 105 bp) was significantly (P < 0.05) higher in 2881 Bos taurus/cross cows (0.82; 95% CI: 0.74, 0.89; I2 = 97.81%) than 1283 Bos indicus cows (0.15; 95% CI: 0.12, 0.18; I2 = 71.90%), with overall gene frequency was 0.33 (95% CI: 0.21, 0.46; I2 = 99.29%). Additive (CC vs. DD) and dominant (CC + CD vs. DD) did not revealed significant (P > 0.05) association with milk yield. However, completely over dominant (CC + DD vs. CD) and recessive (CC vs. CD + DD) models showed significant (P < 0.05) and positive SMDs with milk yield specially at early lactations. There was no evidence of heterogeneity (I2 = 0.00%) between SMDs across studies. This meta-analysis suggested potential association of C allele for enhancing milk production of dairy cows.


The "simulation" of the facial expression of emotions in case of short and long stimulus duration. The effect of pre-motor cortex inhibition by rTMS.
Embodied cognition model states that the "simulation process" is necessary to the recognition of emotional significance of face. The present research explored the contribution of frontal motor brain components (i.e. mainly premotor area) to embodied cognition by using rTMS stimulation, to produce a temporary disruption of this specific cortical site. Secondly, short and long stimulus duration conditions were included to verify the contribution of the "simulation process" in response to overt and covert emotional stimulus comprehension. Nineteen subjects were asked to detect emotion/no emotion (anger, fear, happiness, neutral) in these two experimental conditions, by using a backward masking procedure. Five-second rTMS (1Hz) was delivered before the stimulus onset. False alarms (Fa) and RTs increased and Hits decreased when frontal premotor brain activity was disrupted, specifically in response to anger and fear, for both long and shortduration condition. Thus, the present results highlight the main role of the frontal motor system for emotion facial expression processing.


[Morphological and molecular pathology of lung cancer].
Lung carcinoma is the most frequent cause of cancer death in Germany. It is characterized by a considerable morphological complexity that has been reduced by clinicians to the distinction between small cell (SCLC) and non-small cell carcinoma (NSCLC). Underpinned by the possibilities of more differentiated tumor therapy, the classification of lung cancer has undergone a re-evaluation, some essential developments of which are summarized in this article. SCLC and NSCLC do not only differ in gene expression and genetic alterations but also in the ploidy level: SCLC is typically hypodiploid, NSCLC often hyperdiploid/near-triploid. Immunohistochemical analysis is meanwhile standard and includes in particular the markers p63, TTF-1, CK5/6, CK7, CD56/NCAM, synaptophysin, chromogranin and Ki67. A new interdisciplinary classification of adenocarcinoma differentiates between preinvasive, minimally invasive and invasive lesions. Lending new weight to the predominantly histological growth patterns it includes information on molecular genetic alterations such as EGFR mutations.


Selective tumor localization in experimental hepatoma by radiolabeled antiferritin antibody.
The in vivo localization of 131I-radiolabeled antiferritin and normal IgG in the H-4-II-E rat hepatoma model was investigated by serial necropsy. Groups of 14 to 18 animals were injected with 500 microCi (200 micrograms) of normal and antiferritin IgG. The total dose from the nonpenetrating beta radiation was calculated for tumor and normal tissue, and expressed as a targeting ratio of antiferritin to normal IgG for each organ studied. The results demonstrate 2.9 times greater dose deposition in tumors of animals treated with 131I-antiferritin than with 131I-normal IgG. 131I-antiferritin deposited equivalently in primary tumors and metastatic lesions of similar size. The specific binding in tumors could be competitively inhibited by the addition of unlabeled antiferritin but not unlabeled normal IgG. Specific targeting with 131I-antiferritin comparison to 131I-normal IgG did not occur in any normal tissue. There was considerable variation in the dose deposition in different normal tissues.


Drug Utilization and Medical Cost Study Focusing on Moisturizers in Cancer Patients Treated with Molecular Targeted Therapy: A Retrospective Observational Study Using Data from a Japanese Claims Database.
Molecular targeted therapies (MTTs) cause skin disorders in patients with cancer, and moisturizers are useful treatments; however, their actual use and costs are unknown. Our purpose was to examine the use and costs of moisturizers prescribed for xerosis (asteatosis) in patients with cancer treated with MTTs. We used data from a Japanese hospital-based claims database. The index date was the first date of MTT prescription from October 2011 to April 2018 (selection period), and the follow-up period was 1 year from the index date. Patients treated with MTTs during the selection period and who were not prescribed moisturizers in the 6 months before the index date were included as the study cohort. Timing, duration, amount, and costs of the prescribed moisturizers and total medical costs were analyzed. Among the 78,190 patients in the study cohort, 27,906 patients (35.7%) were prescribed moisturizers during follow-up. Moisturizer prescription timing, duration, and volume were inconsistent. The average annual total medical costs for treating patients with MTT who were prescribed moisturizers was JPY 6.165 million (USD 53,797) per patient, and the moisturizer costs were JPY 6033 (USD 53). The number of patients who used moisturizers showed an increasing trend. No consistent patterns were observed for the timing or duration of moisturizer use, which suggests various developmental patterns of skin disorders. Furthermore, medical costs for moisturizers accounted for only a small proportion of the total medical costs required for cancer treatment.


Cloning of mouse carbonic anhydrase mRNA and its induction in mouse erythroleukemic cells.
Mouse carbonic anhydrase mRNA was detected in poly(A+) RNA of anemic spleens sedimenting as a RNA species at 14 S. Subsequently, poly(A+) RNA (12-16 S) was used as a template for the synthesis of double-stranded cDNA, which was inserted into the PstI site of pBR322 by oligo-dG:dC tailing. A recombinant plasmid containing carbonic anhydrase cDNA was identified by a positive hybridization selection assay and by partial DNA sequencing. Predicted amino acid sequences showed homology with the known sequences of rabbit and human carbonic anhydrase I and II. The clone contained sequences for most of the coding region and 600-700 base pairs at the 3' noncoding region of the mRNA. Hybridization analysis of poly(A+) RNA from uninduced and induced mouse erythroleukemic cells labeled for short and long time periods indicated that induction results in an increase of carbonic anhydrase mRNA in newly synthesized RNA.


Size-dependent vitrification in metallic glasses.
Reducing the sample size can profoundly impact properties of bulk metallic glasses. Here, we systematically reduce the length scale of Au and Pt-based metallic glasses and study their vitrification behavior and atomic mobility. For this purpose, we exploit fast scanning calorimetry (FSC) allowing to study glassy dynamics in an exceptionally wide range of cooling rates and frequencies. We show that the main α relaxation process remains size independent and bulk-like. In contrast, we observe pronounced size dependent vitrification kinetics in micrometer-sized glasses, which is more evident for the smallest samples and at low cooling rates, resulting in more than 40 K decrease in fictive temperature, Tf, with respect to the bulk. We discuss the deep implications on how this outcome can be used to convey glasses to low energy states.


Prenatal diagnosis of arachnoid cysts.
Arachnoid cysts are a rare central nervous system malformation, representing only 1% of all intracranial masses in newborns. Primary (congenital) arachnoid cysts are benign accumulation of clear fluid between the dura and the brain substance throughout the cerebrospinal axis in relation to the arachnoid membrane and do not communicate with the subarachnoid space. Secondary (acquired) arachnoid cysts result from hemorrhage, trauma, and infection and usually communicate with the subarachnoid space. The common locations of arachnoid cysts are the surface of the brain at the level of main brain fissures, such as sylvian, rolandic and interhemispheric fissures, sella turcica, the anterior cranial fossa, and the middle cranial fossa. Arachnoid cysts may be associated with ventriculomegaly and dysgenesis of corpus callosum. Prenatal ultrasound and magnetic resonance imaging have led to the increased diagnosis of fetal arachnoid cysts. This article provides a thorough review of fetal arachnoid cysts, including prenatal diagnosis, differential diagnosis and associated chromosomal abnormalities, as well as comprehensive illustrations of perinatal imaging findings of fetal arachnoid cysts. Prenatal diagnosis of intracranial hypoechoic lesions should include a differential diagnosis of arachnoid cysts and prompt genetic investigations.


[CT in Fournier's gangrene].
To analyze the information provided by computed tomography (CT) in Fournier's gangrene, particularly in regard to the early diagnosis of the disease, and its utility as a complementary diagnostic imaging technique. A retrospective study identified five cases of Fournier's gangrene. These patients had been evaluated by CT at least twice. The radiological, clinical and epidemiological aspects are analyzed. Fournier's gangrene was diagnosed in one patient with no clinical suspicion of this disease. Gas was demonstrated in the scrotal region in five patients and in the perineal region in four patients. The abdominal wall was the most frequent extragenito-perineal location. A good correlation was found between the CT and surgical findings in all cases. CT evaluation prior to surgery is recommended in patients with Fournier's gangrene, particularly in order to determine the extent of the disease.


Exploring personality clusters among parents of ED subjects. Relationship with parents' psychopathology, attachment, and family dynamics.
Eating disorders are some of the most difficult mental disorders to treat and manage. Family interacts with genetic dispositions and other pathogenic factors, and may influence the outburst, development and outcome of EDs. The present study explores with a cluster analysis the personality traits of parents of ED subjects. One-hundred-eight mothers and 104 fathers were tested with Temperament Character Inventory (TCI), Eating Disorder Inventory-2 (EDI-2), State-Trait Anger Expression Inventory (STAX), Family Assessment Device (FAD), Attachment Style Questionnaire (ASQ), Symptom Questionnaire (SQ), Psychological Well-Being scales (PWB). The cluster distribution of parents based on personality traits was explored. Parents' clusters TCI scores were compared as regards personality, psychopathology, attachment and family features. Cross distribution of temperament and character clusters in mothers and fathers, among couples and ED diagnoses of the daughters was explored. Two clusters of mothers and fathers were identified with temperament clustering. Character traits led to two mothers and three fathers clusters. Mothers temperament cluster 1 (MTC1) correspond to a explosive/adventurous profile, MTC2 to a cautious/passive-dependent profile. Fathers temperament cluster 1 (FTC1) was explosive/methodic, FTC2 was independent/methodic. Character clustering distinguished very immature mothers (MCC1) and majority (65%) of character mature mothers with low self-transcendence (MCC2). A third of fathers was severely immature (FCC1), a third impaired as regards relationships (poor cooperativeness and self-transcendence; FCC2), and one third character mature fathers with low self-transcendence (FCC3). Each cluster evidences specific psychopathology and attachment characteristics. FTC1 was more frequently associated with character immaturity. No significant clusters' cross correlation was found in parental couples. Parents' clusters analyze in depth the univocal picture of prototypical mothers and fathers of EDs. Parents not disturbed as regards personality traits are not exceptions. Since EDs are multifactor disorders family dynamics related to parents' personality may be very relevant or even marginal in their pathogenesis. Conversely, parenting may be negatively influenced by relatively marginal personality malfunctions of parents. The clustering approach to the complexity of personality-related dynamics of ED families improves the picture of ED parents. Psychoeducational, counseling and psychotherapeutic family interventions should consider the specific underlying personality of parents.


Immunological biomarkers in severe asthma.
Severe asthma is heterogeneous in its clinical presentation, underlying pathophysiology, course and response to therapy. Clinical and physiological assessment of severe asthma is often inadequate in predicting underlying disease mechanisms and or response to medications. With the emergence of novel targeted therapies in severe asthma, the need for reproducible, easily measured biomarkers became obvious but only few are currently available for clinical use. These biomarkers along with the clinical presentation of the patient play an important role in identifying phenotypes and endotypes, predicting the clinical course and prognosis and improving the precision therapeutic approach to asthma.


A fully automated chromatographic peak detection and treatment software for multi-user multi-task computers.
A fully automated method is presented for the detection and integration of chromatographic peak software in which the only parameter to define is the data acquisition frequency; this has to be chosen high enough to minimize some Nyquist frequency problems. Programmed in C language, the software is used on a Unix like system (Xenix) to emphasize its automaticity. The performance of the system in liquid chromatography has been studied in respect of noise frequency and intensity. It is shown that the classical signal-to-noise ratio is obsolete for definition of the detection limit since frequency analysis allows some signal filtration; these limitations are described. An example is given of simulation with Gaussian peaks convoluted with noise characterized by its intensity and frequency distribution. The validity of the software is demonstrated for some HPLC separations.


Ethosomes® and transfersomes® containing linoleic acid: physicochemical and technological features of topical drug delivery carriers for the potential treatment of melasma disorders.
Two vesicular colloidal carriers, ethosomes® and transfersomes® were proposed for the topical delivery of linoleic acid, an active compound used in the therapeutic treatment of hyperpigmentation disorders, i.e. melasma, which is characterized by an increase of the melanin production in the epidermis. Dynamic light scattering was used for the physicochemical characterization of vesicles and mean size, size distribution and zeta potential were evaluated. The stability of formulations was also evaluated using the Turbiscan Lab® Expert based on the analysis of sample transmittance and photon backscattering. Ethosomes® and transfersomes® were prepared using Phospholipon 100 G®, as the lecithin component, and ethanol and sodium cholate, as edge activator agents, respectively. Linoleic acid at 0.05% and 0.1% (w/v) was used as the active ingredient and entrapped in colloidal vesicles. Technological parameters, i.e. entrapment efficacy, drug release and permeation profiles, were also investigated. Experimental findings showed that physicochemical and technological features of ethosomes® and transfersomes® were influenced by the lipid composition of the carriers. The percutaneous permeation experiments of linoleic acid-loaded ethosomes® and transfersomes® through human stratum corneum-epidermidis membranes showed that both carriers are accumulated in the skin membrane model as a function of their lipid compositions. The findings reported in this investigation showed that both vesicular carriers could represent a potential system for the topical treatment of hyperpigmentation disorders.


Protection of Vigna unguiculata (L.) Walp. plants from salt stress by paclobutrazol.
A pot culture experiment was conducted to estimate the stress ameliorating ability of paclobutrazol, a triazole fungicide in Vigna unguiculata (L.) Walp. plants. Treatments were given as 80 mM NaCl, 80 mM NaCl+15 mg l(-1) paclobutrazol and 15 mg l(-1) paclobutrazol alone. The samples were collected on 60 and 80 days after sowing (DAS). NaCl stress inhibited the root and stem length, total leaf area, fresh weight (FW), dry weight (DW) and activities of antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX). Plants treated with NaCl with paclobutrazol increased these parameters to a larger extent when compared to NaCl stressed plants. The results showed that the paclobutrazol significantly ameliorated the adverse effects of NaCl stress in V. unguiculata plants.


Insights into the functional expansion of the astacin peptidase family in parasitic helminths.
Helminths secrete a plethora of proteins involved in parasitism-related processes such as tissue penetration, migration, feeding and immunoregulation. Astacins, a family of zinc metalloproteases belonging to the peptidase family M12, are one of the most abundantly represented protein families in the secretomes of helminths. Despite their involvement in virulence, very few studies have addressed the role of this loosely defined protein group in parasitic helminths. Herein, we have analysed the predicted proteomes from 154 helminth species and confirmed the expansion of the astacin family in several nematode taxa. The astacin domain associated with up to 110 other domains into 145 unique domain architectures, where CUB and ShK constitute the principal and nearly independent bi-domain frameworks. The presence of co-existing domains suggests promiscuous adaptable functions to several roles. These activities could be related either to substrate specificity or to higher-order functions, such as anti-angiogenesis and immunomodulation, where the astacin domain would play an accessory role. Furthermore, some phylogenetically restricted mutations in the astacin domain affected residues located at the active cleft and binding sub-pockets, suggesting adaptation to different substrate specificities. Altogether, these findings suggest the astacin domain is a highly adaptable module that fulfils multiple proteolytic needs of the parasitic lifestyle. This study contributes to the understanding of helminth-secreted astacins and, ultimately, provides the foundation to guide future investigations about the role of this diverse family of proteins in host-parasite interactions.


Timing of antiretroviral therapy for HIV-infected patients with moderate to severe Pneumocystis pneumonia: study protocol for a multi-centre prospective randomised controlled trial.
Pneumocystis pneumonia (PCP) is a common acquired immune deficiency syndrome (AIDS)-related opportunistic infection. Recent reports estimate that more than 400,000 patients with human immunodeficiency virus (HIV) develop PCP each year globally. However, the timing of antiretroviral therapy (ART) initiation for HIV-infected patients with PCP is still controversial, and the benefits and risks of early initiation of ART are not completely clear. We thus designed this study in order to determine the optimal timing for ART initiation for HIV-positive patients with moderate to severe PCP. This study will be an open-label, multi-centre, prospective randomised controlled trial. A total of 200 subjects will be randomly assigned to an early ART initiation group (≤14 days after PCP diagnosis) and a deferred ART initiation group (>14 days after PCP diagnosis) at a 1:1 ratio. All subjects will be followed up for 48 weeks after starting ART. The primary endpoint is incidence of disease progression (including new or relapsing opportunistic infections and death) at week 48. The secondary endpoints are the changes in CD4 counts from baseline at weeks 12, 24 and 48; the degree of virological suppression (HIV RNA < 50 copies/mL) at weeks 24 and 48; the rate of development of PCP-associated immune reconstitution inflammatory syndrome; and adverse events over 48 weeks. We hope that the results of this study will reveal the optimal timing for initiation of ART in HIV-infected patients with moderate to severe PCP. This trial was registered as one of the 12 trials under the name of a general project at chictr.org.cn on February 1, 2019. The registration number of the general project is ChiCTR1900021195.


Novelty-induced behavioral traits correlate with numbers of brainstem noradrenergic neurons and septal cholinergic neurons in C57BL/6J mice.
It is generally accepted that different brain regions regulate specific behavioral responses and that structural alterations in these regions may affect behavior. We investigated whether inter-individual variability in novelty-induced behaviors in C57BL/6J mice correlates with numbers of noradrenergic neurons in the locus coeruleus and cholinergic neurons in the septum. We found that exploration of new stimuli correlated negatively with numbers of noradrenergic neurons, whereas anxiety correlated positively with numbers of cholinergic neurons. The observed correlations suggest physiologically plausible links between structure and function and indicate that precise morphological estimates can be predictive for behavioral responses.


Immunochemical characteristics of Streptococcus pneumoniae type 3 capsular polysaccharide glycoconjugate constructs correlate with its immunogenicity in mice model.
A panel of derivatives were prepared from Streptococcus pneumoniae polysaccharide type 3 (Ps3) modified with adipic acid dihydrazide (ADH). The degree of coupling between Ps3-adh derivatives and diphtheria (DTd) or tetanus (TTd) toxoids was varied by ADH linker loading. A series of Ps3 derivatives and the resultant glycoconjugates (GC) were tested for their immunochemical activity in an ELISA. Antigenic properties of components in GCs were estimated by interaction with serotype-specific and toxin-neutralizing antibodies to confirm the preservation of native protective epitopes both of Ps3 and DTd. After immunization of mice, a correlation was established between immunochemical activity and immunogenicity of these GCs. A correlation model developed for Ps3-DTd conjugates allowed to predict the immunogenicity of similar design Ps3-TTd conjugates based on ELISA testing data. The plausibility of this prediction was confirmed by the test immunization of mice with Ps3-TTds. The proposed immunochemical approach to the assessment and control of native structural and functional antigenic elements in GCs is important for the optimization of vaccine design and is an adequate alternative to extensive physicochemical characterization for assessing immunogenicity.


Postnatal development of CO2-O2 interaction in the rat carotid body in vitro.
1. The effect of PCO2 upon the discharge response to changes in PO2 (between ca 450 and 30 mmHg) was observed in adult (> 5 weeks old) and neonatal (5-7 days old) rat carotid body chemoreceptors using an in vitro, superfused preparation. 2. In both adult and neonatal rats, regression analysis revealed that increasing PCO2 was without effect upon the shape of the PO2-response curves (P > 0.30), but did cause an upward shift in the position of the curves, as indicated by a significant increase in the baseline chemoreceptor discharge during hyperoxia (0.22 +/- 0.02% maximum discharge per mmHg PCO2, P < 0.001, and 0.25 +/- 0.07% maximum discharge per mmHg PCO2, P < 0.005, respectively). However, whilst increasing PCO2 caused a significant rightward shift of the response curves in adults (0.75 +/- 0.23 mmHg PO2 per mmHg PCO2; P < 0.005), it was without effect in neonates (0.21 +/- 0.22 mmHg PO2 per mmHg PCO2; P > 0.200). Thus increasing levels of hypoxia increased CO2 chemosensitivity in adult but not in neonatal rats as shown by multiple regression analysis of the CO2-response curves which revealed a significant interaction between PCO2 and PO2 for adult (P < 0.010) but not for neonatal (P > 0.150) rats. 3. We suggest that the previously reported maturation of peripheral chemoreceptor hypoxic sensitivity (resetting) may be due to the postnatal emergence of a significant degree of interaction between PCO2 and PO2 at the level of the peripheral chemoreceptor and/or its afferent innervation.


Prevalence and factors associated with sputum smear non-conversion after two months of tuberculosis treatment among smear-positive pulmonary tuberculosis patients in Rwanda: a cross-sectional study.
Non-conversion of sputum smear prolongs the infectivity of pulmonary tuberculosis patients and has been associated with unfavorable tuberculosis (TB) treatment outcomes. Nevertheless, there is a limited evidence on predictors of sputum smear non-conversion among smear-positive PTB (SPPTB) patients in Rwanda. Therefore, this study aimed to determine the factors associated with sputum smear non-conversion after two months of treatment among SPPTB patients in Rwanda. A cross-sectional study was conducted among SPPTB patients registered in the national electronic TB reporting system by all health facilities countrywide (Rwanda) from July 2019 to June 2021. Eligible patients who had completed the first two months of anti-TB treatment and with smear results at the end of the second month of treatment were included in the study. Bivariate and multivariate logistic regression analyses were done using STATA version 16 to determine the factors associated with sputum smear non-conversion. Adjusted odds ratio (OR), 95% confidence interval (CI), and p-value < 0.05 was considered statistically significant. This study included 7,211 patients. Of them, 632 (9%) patients had sputum smear non-conversion at the end of the second month of treatment. In multivariate logistic regression analysis, age groups of 20-39 years (AOR = 1.7, 95% CI: 1.0-2.8) and 40-59 years (AOR:2, 95% CI: 1.1-3.3), history of first-line TB treatment failure (AOR = 2, 95% CI: 1.1-3.6), follow-up by community health workers(CHWs) (AOR = 1.2, 95% CI: 1.0-1.5), BMI < 18.5 at TB treatment initiation (AOR = 1.5, 95% CI: 1.2-1.8), and living in Northern Province of Rwanda (AOR = 1.4, 95% CI: 1.0-2.0), were found to be significantly associated with sputum smear non-conversion after two months of treatment. Sputum smear non-conversion among SPPTB patients remains low in Rwanda compared to countries of similar health care setting. Identified risk factors for sputum smear non-conversion among SPPTB patients in Rwanda were age (20-39 years, 40-59 years), history of first-line TB treatment failure, follow up by CHWs, BMI < 18.5 at TB treatment initiation and residence (Northern province).


Publisher Correction: Real-Time Selective Sequencing with RUBRIC: Read Until with Basecall and Reference-Informed Criteria.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Parallel plate equipment for preparation of uniform gel sheets.
A method for the preparation of uniform gel-disks for enzyme and cell immobilisation, as well as for characterisation of gel mechanical stability, is described. The apparatus comprises a stainless steel base unit and glass parallel plates, designed to permit easy and fast production of multiple homogeneous gel sheets of variable thickness.


Offspring sex ratio in hamsters is correlated with vaginal pH at certain times of mating.
Golden hamsters (Mesocricetus auratus) are sexually receptive for 19-21 h around the time of ovulation. Vaginal pH was measured immediately before mating in four groups of animals each mated at a different time during the female's receptive period. Litter size and sex ratio (percentage male offspring) were recorded at parturition. Mean litter size decreased while offspring sex ratio increased in matings occurring later in estrus. Significant negative correlations were found between vaginal pH and sex ratio of subsequent litters when matings occurred during midestrus. Fluctuations in vaginal pH may differentially affect longevity, motility, or fertilizing capacity of X and Y sperm and/or may reflect other physiological changes in the female which occur over the receptive period. Such a prefertilization mechanism for sex ratio regulation could be highly adaptive since little or no parental reproductive potential is lost.