Using animations to teach biological processes and principles.
Well-designed animations can help students to focus on the underlying principles and processes in biology rather than relying on rote memorization. We present question-driven, terminology-free, "candymation" videos for teaching the concepts behind mitosis and meiosis as an example.


The Spanish standard patch test series: 2016 update by the Spanish Contact Dermatitis and Skin Allergy Research Group (GEIDAC).
The Spanish standard patch test series, as recommended by the Spanish Contact Dermatitis and Skin Allergy Research Group (GEIDAC), has been updated for 2016. The new series replaces the 2012 version and contains the minimum set of allergens recommended for routine investigation of contact allergy in Spain from 2016 onwards. Four haptens -clioquinol, thimerosal, mercury, and primin- have been eliminated owing to a low frequency of relevant allergic reactions, while 3 new allergens -methylisothiazolinone, diazolidinyl urea, and imidazolidinyl urea- have been added. GEIDAC has also modified the recommended aqueous solution concentrations for the 2 classic, major haptens methylchloroisothiazolinone and methylisothiazolinone, which are now to be tested at 200ppm in aqueous solution, and formaldehyde, which is now to be tested in a 2% aqueous solution. Updating the Spanish standard series is one of the functions of GEIDAC, which is responsible for ensuring that the standard series is suited to the country's epidemiological profile and pattern of contact sensitization.


Immediate effects of lumacaftor/ivacaftor administration on lung function in patients with severe cystic fibrosis lung disease.
Safety-data for lumacaftor/ivacaftor (LUM/IVA) combination therapy in patients with severe lung disease (percent predicted forced expiratory volume in 1s [ppFEV1] <40) remain limited. We report immediate post-dose respiratory-related adverse events in 12 patients with severe cystic fibrosis (CF) lung disease (median [IQR] ppFEV1: 34 [31-36]) prescribed LUM/IVA. All patients experienced a decline in ppFEV1 from baseline at 2-hours (median [IQR] relative change: -19 [-21 to -11]%, p<0.001) that persisted at 24-hours but recovered in most patients at 1-month. No pre- and post-differences in bronchodilator response were observed. Ten (83.3%) patients reported non-severe respiratory-related adverse events within 24-hours of LUM/IVA initiation. At 1-month, eight (67%) patients had persistent symptoms and six (50%) were treated for a pulmonary exacerbation. Our results highlight that LUM/IVA respiratory-related adverse events are common in patients with a ppFEV1<40. We recommend close assessment of adverse events. Further studies are required to evaluate the efficacy of LUM/IVA in patients with severe lung disease.


A unique expression pattern of LAG3 distinct from that of other immune checkpoints in diffuse large B-cell lymphoma.
Although some patients with diffuse large B-cell lymphoma (DLBCL) show a response to immunotherapy, there are still many who do not respond. This suggests that various immune checkpoints are complicatedly intertwined in the composition of the tumor microenvironment of DLBCL. To comprehensively understand the expression of various immune checkpoint genes in DLBCL, we performed NanoString assay in 98 patients to investigate 579 genes. In addition, we performed immunohistochemistry for LAG-3 and PD-L1 to compare the results with expression in NanoString assay. As a result of hierarchical clustering of NanoString assay, 98 DLBCLs were classified into three tumor immune microenvironment clusters. Most immune checkpoint genes showed the highest expression in cluster A and the lowest in cluster C. However, the expression of LAG3 was the highest in cluster C and the lowest in cluster A, showing an expression pattern opposite to that of other immune checkpoint genes. In Cluster A, the expression of genes related to T-cell activity such as CD8A and GZMB was increased. In Cluster C, the expression of genes related to major histocompatibility complex molecules was the highest. Immunohistochemical stains showed modest agreement with the NanoString results but did not help clustering. Our results show that the unique expression pattern of LAG3 in DLBCL contrasts with that of other immune checkpoints. We suggest that the combination of anti-PD-1/PD-L1 and anti-LAG-3 blockades in the immunotherapy of DLBCL patients can have a synergistic effect, improving the immunotherapy efficacy and outcome in DLBCL patients.


pH-dependent modulation of TRPV1 by modality-selective antagonists.
Antagonists of TRPV1 that inhibit all activation modes cause hyperthermia, hampering their medical use as novel analgesics. TRPV1 antagonists that do not (fully) inhibit responses to low pH do not cause hyperthermia, but it remains incompletely understood how such antagonists affect channel gating. We tested the hypothesis that pH-sparing antagonists act in a modality-selective manner on TRPV1, differentially affecting channel activation by protons and capsaicin. Using whole-cell patch-clamp and calcium imaging to measure channel activity in cells expressing wild type human TRPV1 or the pH-insensitive mutant F660A. Responses to protons and capsaicin were measured at different pH values in the presence of antagonists that reportedly partially spare (A-1165442) or potentiate (AMG7905) acid-evoked channel activation. At pH 5.5, A-1165442 was equipotent at blocking acid- and capsaicin-evoked responses of wild type TRPV1. Its potency to inhibit acid-evoked responses was attenuated at pH ≤ 5.0. AMG7905, at a concentration (1 μM) that fully inhibits capsaicin-evoked responses, potentiated proton-evoked (pH 5.5) responses of wild type TRPV1. In the F660A mutant, the inhibitory efficacy of A-1165442 and AMG7905 towards capsaicin-evoked responses was reduced at lower pH values and AMG7905 acted as a partial agonist. Our findings show that A-1165442 and AMG7905 interact in a pH-dependent manner with TRPV1, but this pH dependence is not strictly modality-selective. Reduced TRPV1 antagonism at acidic pH may limit analgesic efficacy in injured tissue and needs to be considered in models explaining the effects of antagonists on core body temperature.


Adjustable inspiratory occlusion valve in experimental bronchopleural fistula. A new therapeutic perspective.
To assess the efficacy of an adjustable inspiratory occlusion valve in experimental bronchopleural fistula during mechanical ventilation. We studied six mechanically ventilated pigs in a surgically created, reproducible model of bronchopleural fistula managed with mechanical ventilation and water-sealed thoracic drainage. An adjustable inspiratory occlusion valve was placed between the thoracic drain and the endotracheal tube. Hemodynamic data, capnography and blood gases were recorded before and after the creation of the bronchopleural fistula as well as after every adjustment of the inspiratory occlusion valve. When compared with the standard water-sealed drainage treatment, the use of an adjustable inspiratory occlusion valve improved the alveolar tidal volume and reduced bronchopleural air leak (p<0.001), without hemodynamic compromise when compared with conventional water sealed drainage. The use of an adjustable inspiratory occlusion valve improved the alveolar tidal volume, reduced alveolar leak, in an experimental reproducible model of bronchopleural fistula, without causing any hemodynamic derangements when compared with conventional water sealed drainage.


Experimental and computational framework for a dynamic protein atlas of human cell division.
Essential biological functions, such as mitosis, require tight coordination of hundreds of proteins in space and time. Localization, the timing of interactions and changes in cellular structure are all crucial to ensure the correct assembly, function and regulation of protein complexes1-4. Imaging of live cells can reveal protein distributions and dynamics but experimental and theoretical challenges have prevented the collection of quantitative data, which are necessary for the formulation of a model of mitosis that comprehensively integrates information and enables the analysis of the dynamic interactions between the molecular parts of the mitotic machinery within changing cellular boundaries. Here we generate a canonical model of the morphological changes during the mitotic progression of human cells on the basis of four-dimensional image data. We use this model to integrate dynamic three-dimensional concentration data of many fluorescently knocked-in mitotic proteins, imaged by fluorescence correlation spectroscopy-calibrated microscopy5. The approach taken here to generate a dynamic protein atlas of human cell division is generic; it can be applied to systematically map and mine dynamic protein localization networks that drive cell division in different cell types, and can be conceptually transferred to other cellular functions.


Prader-Willi syndrome.
Prader-Willi syndrome is characterized by severe infantile hypotonia with poor suck and failure to thrive; hypogonadism causing genital hypoplasia and pubertal insufficiency; characteristic facial features; early-childhood onset obesity and hyperphagia; developmental delay/mild intellectual disability; short stature; and a distinctive behavioral phenotype. Sleep abnormalities and scoliosis are common. Growth hormone insufficiency is frequent, and replacement therapy provides improvement in growth, body composition, and physical attributes. Management is otherwise largely supportive. Consensus clinical diagnostic criteria exist, but diagnosis should be confirmed through genetic testing. Prader-Willi syndrome is due to absence of paternally expressed imprinted genes at 15q11.2-q13 through paternal deletion of this region (65-75% of individuals), maternal uniparental disomy 15 (20-30%), or an imprinting defect (1-3%). Parent-specific DNA methylation analysis will detect >99% of individuals. However, additional genetic studies are necessary to identify the molecular class. There are multiple imprinted genes in this region, the loss of which contribute to the complete phenotype of Prader-Willi syndrome. However, absence of a small nucleolar organizing RNA gene, SNORD116, seems to reproduce many of the clinical features. Sibling recurrence risk is typically <1%, but higher risks may pertain in certain cases. Prenatal diagnosis is available.


Lifelong docosahexaenoic acid intervention ameliorates aging in the telomere-DNA-mitochondria axis in telomerase-deficient mice.
The health benefits of n-3 polyunsaturated fatty acids (PUFAs) in multiple age-related diseases are associated with telomere length. Telomerase is intimately related to inflammation and oxidative stress, but whether the underlying function of n-3 PUFAs on telomere maintenance is based on telomerase activation or related mechanisms remains unclear. Herein, we utilized late-generation (G4) telomerase-deficient (Terc-/-) mice to perform a lifelong docosahexaenoic acid (DHA) intervention to determine the potential of DHA in telomere maintenance and health promotion. Unfortunately, DHA failed to prolong mouse longevity in either intrinsic or premature aging. However, intriguingly, lifelong dietary DHA intervention slowed the aging phenotypes and profoundly attenuated telomere attrition in blood leukocytes and multiple tissues, consistent with decreased β-galactosidase activity and other senescence hallmarks with no observed sex differences. Notably, DHA intervention alleviated telomere attrition-induced γ-H2AX accumulation dependent on poly (ADP-ribose) polymerase 1 (PARP1) recruitment, and further regulated mitochondrial dysfunction critically involved in the DNA damage response. Together with the improvement of mitochondria function, the blocked reactive oxygen species (ROS) accumulation and suppression of the nuclear factor-κB (NF-κB)/nucleotide-binding domain-like receptor protein 3 (NLRP3)/caspase-1 pathways partially indicated anti-oxidative and anti-inflammatory effects of DHA. These data revealed a regulatory paradigm involving DHA in the telomere-DNA-mitochondria feedback loop mediated by DNA damage response and inflammation in alleviating senescence, which may hold potential as a translatable intervention in telomere-related diseases during aging.


Genetic Diversity, Repeat Motifs, and Natural Selection at the C-Terminal Knob-Associated Histidine Rich Protein (KAHRP) of Plasmodium falciparum Clinical Samples from Saudi Arabia.
Malaria is still a public health problem in Saudi Arabia specifically in the Jazan region. Plasmodium falciparum knob-associated histidine-rich proteins (PfKAHRPs) play an important role in cerebral malaria pathophysiology as well as pathogenesis of P. falciparum infections. The repeat region of PfKAHRP C-terminal interaction domain has been found to bind to the infected red blood cells and the vascular endothelium. Thus, this study aimed to assess the allelic variations, genetic diversity, and natural selection acting at the C-terminal PfKAHRP between parasite isolates from Saudi Arabia. The PfKHARP C-terminal interaction domain was successfully PCR-amplified and sequence data from 441 clinical isolates from Saudi Arabia were obtained. The DnaSP v5.10 software was used to determine the genetic diversity, polymorphism, haplotype, and natural selection. Haplotype network analysis was constructed by using the median-joining method in the NETWORK version 184.108.40.206 software. Alignment and analysis of 441 C-terminal PfKAHRP-deduced amino acid sequences identified 5 genotypes (I-V) based on the decapeptide repeat arrangements (TKEASTSKEA, TKEASTSKGA, TKEASTTEGA, and TKEASTSKRA). Among the repeat types, Type I (49.43%, 218/441) was the most abundant in Saudi Arabia, followed by Type II (48.29%, 213/441). Overall, the nucleotide diversity in the PfKHARP C-terminal region was found to be low in Saudi Arabia (π = 0.00142); however, natural selection tests indicated positive selection (dN-dS = 1.64, P < 0.05) which was due to the variations within the repeat motifs. Genealogical relationship haplotype network of PfKAHRP from 4 different countries (i.e., Saudi Arabia, Iran, Burundi, and India) revealed 1 major shared haplotype cluster (H_1) with samples representative from all 4 countries (Saudi Arabia; n = 441, Burundi; n = 4, Iran; n = 13, and India; n = 1). Since this is the first study to report on genetic diversity of C-terminal PfKAHRP interaction domain and the repeat motifs from clinical samples in Saudi Arabia, it will contribute towards the rational design of antiadhesion drug therapies for P. falciparum malaria.


Effect of Chromium on Corrosion Behavior of P110 Steels in CO₂-H₂S Environment with High Pressure and High Temperature.
The novel Cr-containing low alloy steels have exhibited good corrosion resistance in CO₂ environment, mainly owing to the formation of Cr-enriched corrosion film. In order to evaluate whether it is applicable to the CO₂ and H₂S coexistence conditions, the corrosion behavior of low-chromium steels in CO₂-H₂S environment with high pressure and high temperature was investigated using weight loss measurement and surface characterization. The results showed that P110 steel suffered localized corrosion and both 3Cr-P110 and 5Cr-P110 steels exhibited general corrosion. However, the corrosion rate of 5Cr-P110 was the highest among them. The corrosion process of the steels was simultaneously governed by CO₂ and H₂S. The outer scales on the three steels mainly consisted of FeS1-x crystals, whereas the inner scales on Cr-containing steels comprised of amorphous FeS1-x, Cr(OH)₃ and FeCO₃, in contrast with the amorphous FeS1-x and FeCO₃ mixture film of P110 steel. The more chromium the steel contains, the more chromium compounds the corrosion products contain. The addition of chromium in steels increases the uniformity of the Cr-enriched corrosion scales, eliminates the localized corrosion, but cannot decrease the general corrosion rates. The formation of FeS1-x may interfere with Cr-enriched corrosion scales and lowering the corrosion performance of 3Cr-P110 and 5Cr-P110 steels.


Stop Hypertension with the Acupuncture Research Program (SHARP): clinical trial design and screening results.
Hypertension is a major public health problem with serious medical and financial consequences. Barriers to successful conventional pharmacological treatment include side effects, out-of-pocket expenses, patient noncompliance and insufficient dosages. Acupuncture has been studied as an alternative therapy for controlling blood pressure (BP) but previous studies have serious methodological limitations. This paper describes the design of the Stop Hypertension with the Acupuncture Research Program (SHARP) trial, a pilot randomized clinical trial designed to gather preliminary data regarding the efficacy of traditional Chinese medicine (TCM)-based acupuncture for control of essential hypertension. The design of the SHARP trial balanced rigorous clinical trial methodology with principles of TCM. Eligible participants had systolic BP (SBP) 140-179 mm Hg and diastolic BP (DBP) 90-109 mm Hg in the absence of antihypertensive therapy. Following screening, participants were randomized to one of three groups: individualized, standardized or control acupuncture. Treatments were designed according to principles of TCM; nonspecific effects associated with the interventions were standardized across the randomized groups. For individualized acupuncture, points were tailored to each participant. Standardized acupuncture used a prespecified set of points. The invasive sham control acupuncture regimen was designed to be non-active. Each participant received a "prescription" for individualized acupuncture from an acupuncturist who was masked to treatment assignment, and was subsequently treated by an independent acupuncturist. Patients and those assessing BP were masked to treatment group. Acupuncture was delivered twice a week for 6 weeks. Follow-up visits were every 2 weeks to week 10 and then at months 4, 6, 9 and 12. The primary endpoint will be change in SBP from baseline to 10 weeks. DBP, BP trajectories over the 12-month follow-up and antihypertensive medication requirements will also be examined. Initial contact was documented for 1442 prospective participants from March 2001 to April 2002; 424 provided informed consent and 192 were ultimately randomized.


Inhibition of oxidative phosphorylation by palmitoyl-CoA in digitonin permeabilized fibroblasts: implications for long-chain fatty acid beta-oxidation disorders.
Long-chain fatty acid oxidation deficient patients present early in life with more severe features than patients with a medium-chain fatty acid oxidation deficiency. This may be related to the more toxic effect of long-chain fatty acid derivatives. In this paper we have studied the effect of different acyl-CoA esters, and palmitoyl-CoA in particular, on succinate-driven oxidative phosphorylation, using digitonin permeabilized human fibroblasts. Palmitoyl-CoA was found to inhibit the succinate-driven oxidative phosphorylation in a concentration dependent manner. If the inhibition of the oxidative phosphorylation system is also expressed under in vivo conditions this might explain some of the abnormalities found in patients with defects in long-chain fatty acid beta-oxidation.


Active integrins regulate white adipose tissue insulin sensitivity and brown fat thermogenesis.
Reorganization of the extracellular matrix is a prerequisite for healthy adipose tissue expansion, whereas fibrosis is a key feature of adipose dysfunction and inflammation. However, very little is known about the direct effects of impaired cell-matrix interaction in adipocyte function and insulin sensitivity. The objective of this study was to determine whether integrin activity can regulate insulin sensitivity in adipocytes and thereby systemic metabolism. We characterized integrin activity in adipose tissue and its consequences on whole-body metabolism using adipose-selective deletion of β1 integrin (Itgb1adipo-cre) and Kindlin-2 (Kind2adipo-cre) in mice. We demonstrate that integrin signaling regulates white adipocyte insulin action and systemic metabolism. Consequently, loss of adipose integrin activity, similar to loss of adipose insulin receptors, results in a lipodystrophy-like phenotype and systemic insulin resistance. However, brown adipose tissue of Kind2adipo-cre and Itgb1adipo-cre mice is chronically hyperactivated and has increased substrate delivery, reduced endothelial basement membrane thickness, and increased endothelial vesicular transport. Thus, we establish integrin-extracellular matrix interactions as key regulators of white and brown adipose tissue function and whole-body metabolism.


Is the interpretation of rapid antigen testing for respiratory syncytial virus as simple as positive or negative?
To measure the performance characteristics of an immunochromatographic rapid antigen test for respiratory syncytial virus (RSV) and determine how its interpretation should be contextualised in patients presenting to the emergency department (ED) with bronchiolitis. Diagnostic accuracy study of a rapid RSV test. County hospital ED. We took paired nasal samples from consecutively enrolled infants with bronchiolitis and tested them with a rapid immunochromatographic antigen test and reverse transcriptase PCR gold standard. Sensitivity, specificity, the effect of point prevalence, clinical findings and overall context on predictive values. We used these to construct a graphical contextual model to show how the results of RSV antigen tests from infants presenting within 24 h should influence interpretation of subsequent antigen tests. We analysed 607 patients. The sensitivity and specificity for immunochromatographic testing was 79.4% (95% CI 73.9% to 84.2%) and 67.1% (95% CI 61.9% to 72%) respectively. We found little evidence of spectrum bias. In our contextual model the best predictor of a positive RT-PCR test was a positive antigen test OR 5.47 (95% CI 3.65 to 8.18) and the number of other infants having positive tests within 24 h OR 1.48 (95% CI 1.26 to 1.72) per infant. Increasing numbers presenting to the ED with bronchiolitis in a given day increases the probability of RSV infection. The RSV antigen test we examined had modest performance characteristics. The results of the antigen test should be interpreted in the context of the results of previous tests.


Axodendritic and dendrodendritic synapses within outer spiral bundles in a human.
Axodendritic and dendrodendritic synapses have been described at the level of the outer spiral bundle (OSB) (Nadol, J.B., Jr., 1983. Laryngoscope 93, 780-791; Bodian, D., 1978. Proc. Natl. Acad. Sci. USA 75, 4582-4586). The objectives of this study were to quantify these synaptic interactions and to describe their ultrastructural morphology in a young human subject. The temporal bone of an 8-month old infant was processed for transmission electron microscopy and semiserial section reconstructions of the three OSBs were performed. The nerve fibers ((NFs)) forming the OSBs were found to segregate into two morphological groups: (1) vesicle-rich and neurofilament-poor (VR/NP); (2) vesicle-poor and neurofilament-rich (VP/NR). Synapses between VR/NP and VP/NR NFs and synapses between two VP/NR NFs were quantified. Presumed axodendritic synapses (i.e. between VR/NP and VP/NR NFs) were numerous and their numbers decreased from the first towards the third row. Presumed dendrodendritic synapses (i.e. between two VP/NR NFs) were also frequent but their numbers did not vary significantly among different rows. The presence of axodendritic synapses may provide the morphological basis for modulation of the function of the type II spiral ganglion cells (type II's) by the olivocochlear efferent system. Similarly, numerous presumed dendrodendritic synapses may provide a morphological substrate for interaction between dendrites of type II's.


Gestational age-specific reference intervals for blood copper, zinc, calcium, magnesium, iron, lead, and cadmium during normal pregnancy.
The aim of this study was to investigate the blood copper (Cu), zinc (Zn), calcium (Ca), magnesium (Mg), iron (Fe), lead (Pb), and cadmium (Cd) concentrations in women of ≤12 (group I), 13-20 (group II), 21-27 (group III), 28-35 (group IV), and 36-42 (group V) weeks of gestation and compare them with those in nonpregnant women. The cross-sectional study was performed in 2380 pregnant women [group I (n = 550); group II (n = 552); group III (n = 600); group IV (n = 553); and group V (n = 125)] and 552 nonpregnant women as controls. Blood seven element concentrations, including Cu, Zn, Ca, Mg, Fe, Pd, and Cd, were determined by atomic absorption spectrometry (AAS). Compared with the nonpregnant women group, the concentrations of Cu, Zn, Ca, Mg, Fe, Pb, and Cd at ≤12, 13-20, 21-27, 28-35, and 36-42 weeks of gestation, on the whole, were significantly different. Blood Cu, Mg, Ca, Fe, Pb, and Cd concentrations were correlated with weeks of gestation (P<0.05). The gestational age-specific reference intervals were established for Cu, Zn, Ca, Mg, Fe, Pd, and Cd. The established reference intervals for Cu, Zn, Ca, Mg, and Fe can provide important guidance for the reasonable supplementation of essential elements in pregnancy. And the reference intervals for Pd and Cd can play an important part in the surveillance and diagnosis of environmental overexposure.


SCE increases after an accidental acute inhalation exposure to EtO and recovery to normal after 2 years.
Sister-chromatid exchanges (SCEs) were studied in 3 workers accidentally exposed for about half an hour to high levels of ethylene oxide (more than 700 ppm) during a sterilizing process. The 3 workers had clinical symptoms of exposure and were tested for SCE frequencies 5 days and 2 years after the accident: 2 had also been tested 6 months earlier. All 3 showed a similar increase in SCEs after the accident, to a mean of 13.8 SCEs/cell compared with 8.6 for a control group. The incidence of 'high-frequency cells' was markedly elevated but decreased over 2 years, when the mean SCE frequencies had also returned to pre-accident levels.


[Structural characteristics and oxidative phosphorylation of the isolated mitochondria from the rabbit small intestine epithelium under normal conditions and under the effect of choleragen].
Mitochondrial fraction was isolated from the rabbit small intestine epithelium by differential centrifugation in isotonic sucrose medium. The fraction was insignificantly contaminated with fragments of other organellae and microvilli. Respiratory control coefficient of such mitochondria constituted 3-5, when malate-glutamate mixture was used as a substrate. Changes in the functional state of mitochondria were accompanied by stereotypical reconstruction of the "orthodox-condensed" configuration. Introduction of crude choleragen into the mitochondria incubation medium failed to induce any changes in the rate of oxygen consumption in states III and IV (after Chance).


Factors affecting the chemical efficacy of 2% sodium hypochlorite against oral steady-state dual-species biofilms: Exposure time and volume application.
To study the influence of time and volume of 2% sodium hypochlorite (NaOCl) on biofilm removal and to investigate the changes induced on the biofilm architecture. Steady-state, dual-species biofilms of standardized thickness and a realistic contact surface area between biofilms and NaOCl were used. Streptococcus oralis J22 and Actinomyces naeslundii T14V-J1 biofilms were grown on saliva-coated hydroxyapatite discs within sample holders in the Constant Depth Film Fermenter (CDFF) for 96 h. Two per cent NaOCl was statically applied for three different time intervals (60, 120 and 300 s) and in two different volumes (20 and 40 μL) over the biofilm samples. The diffusion-driven effects of time and volume on biofilm disruption and dissolution were assessed with Optical Coherence Tomography (OCT). Structural changes of the biofilms treated with 2% NaOCl were studied with Confocal Laser Scanning Microscopy (CLSM) and Low Load Compression Testing (LLCT). A two-way analysis of variance (2-way anova) was performed, enabling the effect of each independent variable as well as their interaction on the outcome measures. Optical coherence tomography revealed that by increasing the exposure time and volume of 2% NaOCl, both biofilm disruption and dissolution significantly increased. Analysis of the interaction between the two independent variables revealed that by increasing the volume of 2% NaOCl, significant biofilm dissolution could be achieved in less time. Examination of the architecture of the remaining biofilms corroborated the EPS-lytic action of 2% NaOCl, especially when greater volumes were applied. The viscoelastic analysis of the 2% NaOCl-treated biofilms revealed that the preceding application of higher volumes could impact their subsequent removal. Time and volume of 2% NaOCl application should be taken into account for maximizing the anti-biofilm efficiency of the irrigant and devising targeted disinfecting regimes against remaining biofilms.


On calculating sterility in thermal preservation methods: application of the Weibull frequency distribution model.
A simple and parsimonious model which originated from the Weibull frequency distribution was proposed to describe nonlinear survival curves of spores. This model was suitable for downward concavity curves (Bacillus cereus and Bacillus pumilus), as well as for upward concavity curves (Clostridium botulinum). It was shown that traditional F values calculated from this new model were no longer additive, to such an extent that a heat treatment should be better characterized by the obtained decimal reduction of spores. A modified Bigelow method was then proposed to assess this decade reduction or to optimize the heat treatment for a target reduction ratio.


Roux-en-Y gastric bypass, gastric banding, or sleeve gastrectomy for severe obesity: Baseline data from the By-Band-Sleeve randomized controlled trial.
This paper reports the study design, participant characteristics, and recruitment results of By-Band-Sleeve, which investigated the clinical and cost-effectiveness of gastric bypass, gastric banding, and sleeve gastrectomy in adults with severe obesity in the UK. A pragmatic open adaptive noninferiority trial with 3-year follow-up was conducted. Participants were randomly assigned to bypass or band initially and to sleeve after the adaptation. Co-primary end points are weight loss and health-related quality of life assessed using the EQ-5D utility index. Between December 2012 and August 2015, the study recruited into two groups and, after the adaptation, into three groups until September 2019. The study screened 6960 patients; 4732 (68%) were eligible and 1351 (29%) were randomized; 5 subsequently withdrew consent to use data, leaving 462, 464, and 420 assigned to bypass, band, and sleeve, respectively. Baseline data showed high levels of obesity (mean BMI = 46.4 kg/m2; SD: 6.9) and comorbidities (e.g., 31% diabetes), low scores for health-related quality of life, and high levels of anxiety and depression (e.g., 25% abnormal scores). Nutritional parameters were poor, and the average equivalized household income was low (£16,667). By-Band-Sleeve fully recruited. Participant characteristics are consistent with contemporary patients having bariatric surgery, and therefore the results will be generalizable.


[Tarsorrhaphy in rehabilitation of patients with thyroid eye disease].
To evaluate the role of tarsorrhaphy and its effectiveness in the treatment of patients with thyroid eye disease (TED). The study identified the patients who required tarsorrhaphy for corneal lesions associated with TED among the total number of 457 patients who underwent tarsorrhaphy at the Research Institute of Eye Diseases over the past 20 years. Overall, 477 tarsorrhaphy surgeries (457 patients) were performed, including temporary and permanent, partial and complete. The corneal involvement in TED was the indication for tarsorrhaphy in 81 patients (101 tarsorrhaphies). TED patients were divided depending on the date of operation, and it was found that 61 tarsorrhaphies were performed in the period from 2000 to 2009, and 40 similar interventions were performed in the period from 2010 to 2019. The reduction in the number of tarsorrhaphy surgeries in patients with TED in the last decade due to improvement of techniques for bony decompression of the orbit and increase in the number of these operations allowed significant reduction of the degree of proptosis and, as a consequence, the exposure surface area of the eye. However, despite all the successes achieved in orbital surgery, tarsorrhaphy still remains an affordable technique that can always be used in case of corneal lesions in thyroid eye disease.


[Elbow replacement].
Elbow replacement or arthroplasty is a good therapeutic option for a large percentage of patients with significant joint destruction. However, many orthopaedic surgeons are no familiar with the surgical approaches or techniques associated with elbow replacement implants. Furthermore, the incidence of complications is higher than in other joint replacements, the most important being infections, mechanical failure, cubital neuropathy, and problems with the triceps. For these reasons, the use of bone arthroplasty in Spain may be less than ideal. Although, inflammatory arthritic diseases, such as rheumatoid arthritis, are the most frequent indication for this operation, distal humerus fractures and post-traumatic disease are a growing indication. This work attempts to summarise the most important current concepts associated with elbow replacement.


Nasopharyngeal cancer in a total population: selected clinical and epidemiological aspects.
A nationwide study of nasopharyngeal cancer in Israel, during a 9-year period (1960-68) demonstrated a mean annual incidence rate of 1.0 per 100,000 in males and 0.4 per 100,000 in females. Significantly higher incidence was observed in residents born in North Africa (3.0 in males and 1. 1 in females). Survival was relatively better in females among patients with lymphoepithelioma and in those without neurological complications. The results may support the presence of environmental factors in aetiology, though genetic predisposition cannot be ruled out.


Equillibrium disorders with diffuse brain atrophy in long-term toluene sniffing.
A Japanese male who became habituated to sniffing toluene exhibited ataxia, tremor, incoordination and equilibrium disorders. There was pendular nystagmus with a normal caloric response. There were an inhibition of optokinetic nystagmus and a failure to follow over 0.1 Hz sine waves on eye tracking test. Angiogram and pneumoencephalogram revealed an atrophy of the midbrain and cerebrum, and degeneration of the cerebellum was suspected.


Memory-enhancing effects of post-training dipivefrin and epinephrine: involvement of peripheral and central adrenergic receptors.
These experiments examined the effects, in mice, of post-training i.p. injections of dipivefrin (DPE), a lipophilic prodrug of epinephrine, and epinephrine (EPI) on 48-h retention assessed in inhibitory avoidance and Y-maze discrimination tasks. DPE, in doses of 0.3-10 micrograms/kg significantly facilitated retention: the effects were approximately 10-fold more potent than those of EPI obtained with similar experimental conditions. The alpha-adrenergic antagonists prazosin (alpha 1; 3.0 mg/kg; i.p.), yohimbine (alpha 2; 3.0 mg/kg; i.p.) and phentolamine (alpha 1 and alpha 2; 3.0 mg/kg; i.p.) did not block the enhancement of retention induced by either DPE (10.0 micrograms/kg; i.p.) or EPI (0.1 mg/kg; i.p.). However, the beta-adrenergic antagonist propranolol (2.0 mg/kg; i.p.) attenuated the effects of both DPE and EPI. Sotalol (2.0 mg/kg; i.p.), a peripherally-acting beta-adrenergic antagonist, attenuated the effects of EPI but not those of DPE. These findings suggest the DPE-induced enhancement of memory involves central beta- but not alpha-adrenergic mechanisms while EPI's effects are initiated by activation of peripheral beta-adrenergic systems.


Bariatric Surgery in Medicare Patients: Examining Safety and Healthcare Utilization in the Disabled and Elderly.
To compare safety and healthcare utilization after sleeve gastrectomy versus Roux-en-Y gastric bypass in a national Medicare cohort. Though bariatric surgery is increasing among Medicare beneficiaries, no long-term, national studies examining comparative effectiveness between procedures exist. Bariatric outcomes are needed for shared decision-making and coverage policy concerns identified by the cMS Medicare Evidence Development and Coverage Advisory Committee. Retrospective instrumental variable analysis of Medicare claims (2012-2017) for 30,105 bariatric surgery patients entitled due to disability or age. We examined clinical safety outcomes (mortality, complications, and reinterventions), healthcare utilization [Emergency Department (ED) visits, rehospitalizations, and expenditures], and heterogeneity of treatment effect. We compared all outcomes between sleeve and bypass for each entitlement group at 30 days, 1 year, and 3 years. Among the disabled (n = 21,595), sleeve was associated with lower 3-year mortality [2.1% vs 3.2%, absolute risk reduction (ARR) 95% confidence interval (CI): -2.2% to -0.03%], complications (22.2% vs 27.7%, ARR 95%CI: -8.5% to -2.6%), reinterventions (20.1% vs 27.7%, ARR 95%CI: -10.7% to -4.6%), ED utilization (71.6% vs 77.1%, ARR 95%CI: -8.5% to -2.4%), and rehospitalizations (47.4% vs 52.3%, ARR 95%Ci: -8.0% to -1.7%). Cumulative expenditures were $46,277 after sleeve and $48,211 after bypass (P = 0.22). Among the elderly (n = 8510), sleeve was associated with lower 3-year complications (20.1% vs 24.7%, ARR 95%CI: -7.6% to -1.7%), reinterventions (14.0% vs 21.9%, ARR 95%CI: -10.7% to -5.2%), ED utilization (51.7% vs 57.2%, ARR 95%CI: -9.1% to -1.9%), and rehospitalizations (41.8% vs 45.8%, ARR 95%Ci: -7.5% to -0.5%). Expenditures were $38,632 after sleeve and $39,270 after bypass (P = 0.60). Procedure treatment effect significantly differed by entitlement for mortality, revision, and paraesophageal hernia repair. Bariatric surgery is safe, and healthcare utilization benefits of sleeve over bypass are preserved across both Medicare elderly and disabled subpopulations.


A review of commonly performed bariatric surgeries: Imaging features and its complications.
Obesity is a disease that has achieved the level that can be considered an epidemic. According to the National Center for Health Statistics data, the prevalence of obesity has increased from 30.5% in 1999-2000 to 42.4% in 2017-2018. During the same period, severe obesity has increased from 4.7% to 9.2%. With the growing prevalence of obesity, related conditions such as coronary artery disease, diabetes, and strokes have also become more prevalent. In the past few years, the need for bariatric surgeries such as laparoscopic Roux-en-Y gastric bypass, sleeve gastrectomy, and laparoscopic adjustable gastric banding has increased considerably. With an increasing number of bariatric surgeries, multiple postoperative complications have become common. In this review, we have attempted to describe normal postsurgical anatomical findings after bariatric surgeries and pictorial review of a few common postoperative complications.


Re: The correlation between PD-L1 expression and metabolic parameters of 18FDG PET/CT and the prognostic value of PD-L1 in non-small cell lung cancer.
We read with great interest the recent study conducted by Xu et al1. In this study, the authors investigated the correlation of programmed death ligand-1 (PD-L1) expression with metabolic parameters measured by 18F-fluorodeoxyglucose positron emission computed tomography (18F-FDG PET/CT) and its prognostic value in patients with surgically resected non-small-cell lung cancer (NSCLC). The results of this study1 found that in NSCLC patients, the maximum standardized uptake value (SUVmax) measured by 18F-FDG PET/CT was a predictor of PD-L1 expression, and both PD-L1 and TLG were prognostic factors for the prognosis of NSCLC patient. Undoubtedly, the novel findings of this study help to further refine the risk stratification of NSCLC patients. However, after reading this article carefully, we would like to share the following points.


Comparative studies on thymidylate synthetase from P. berghei and mouse reticulocytes.
Partially purified thymidylate synthetase from Plasmodium berghei and mouse reticulocytes was characterized. The mol. wt of the enzyme from P. berghei was about twice that from mouse reticulocytes. The optimum pH of the enzyme from P. berghei was found to be 6.5-7.5 while that from the host was 7.0-8.0. The enzyme from P. berghei was more susceptible to pH denaturation than the enzyme from reticulocytes. The enzyme from both sources differed in their Km values for substrates. The enzyme from reticulocytes was less sensitive to inhibition by substrate analogs than that from P. berghei.


Polarization-resolved SHG imaging as a fast screening method for collagen alterations during aging: Comparison with light and electron microscopy.
Our previous study on rat skin showed that cumulative oxidative pressure induces profound structural and ultrastructural alterations in both rat skin epidermis and dermis during aging. Here, we aimed to investigate the biophotonic properties of collagen as a main dermal component in the function of chronological aging. We used second harmonic generation (SHG) and two-photon excited fluorescence (TPEF) on 5 μm thick skin paraffin sections from 15-day-, 1-month- and 21-month-old rats, respectively, to analyze collagen alterations, in comparison to conventional light and electron microscopy methods. Obtained results show that polarization-resolved SHG (PSHG) images can detect collagen fiber alterations in line with chronological aging and that this method is consistent with light and electron microscopy. Moreover, the β coefficient calculated from PSHG images points out that delicate alterations lead to a more ordered structure of collagen molecules due to oxidative damage. The results of this study also open the possibility of successfully applying this fast and label-free method to previously fixed samples.


Leaf anatomy and light acclimation in woody seedlings after gap formation in a cool-temperate deciduous forest.
The photosynthetic light acclimation of fully expanded leaves of tree seedlings in response to gap formation was studied with respect to anatomical and photosynthetic characteristics in a natural cool-temperate deciduous forest. Eight woody species of different functional groups were used; two species each from mid-successional canopy species (Kalopanax pictus and Magnolia obovata), from late-successional canopy species (Quercus crispula and Acer mono), from sub-canopy species (Acer japonicum and Fraxinus lanuginosa) and from vine species (Schizophragma hydrangeoides and Hydrangea petiolaris). The light-saturated rate of photosynthesis (Pmax) increased significantly after gap formation in six species other than vine species. Shade leaves of K. pictus, M. obovata and Q. crispula had vacant spaces along cell walls in mesophyll cells, where chloroplasts were absent. The vacant space was filled after the gap formation by increased chloroplast volume, which in turn increased Pmax. In two Acer species, an increase in the area of mesophyll cells facing the intercellular space enabled the leaves to increase Pmax after maturation. The two vine species did not significantly change their anatomical traits. Although the response and the mechanism of acclimation to light improvement varied from species to species, the increase in the area of chloroplast surface facing the intercellular space per unit leaf area accounted for most of the increase in Pmax, demonstrating the importance of leaf anatomy in increasing Pmax.


Soluble antigen of M. leprae coupled with liposomes elicits both "early" and "late" delayed hypersensitivity skin reactions.
The soluble antigen(s) of Mycobacterium leprae was(were) coupled to liposomes and used for skin testing of leprosy patients, hoping that this mode of antigen presentation would be identical to that of integral lepromin. The liposomized antigen(s) elicited both early (24-48 hr) and late (3-4 weeks) delayed-type hypersensitivity reactions, true to the nature of lepromin, unlike the soluble antigen(s) alone which elicit(s) only the early reaction.


Human dermal fibroblast subpopulations; differential interactions with vascular endothelial cells in coculture: nonsoluble factors in the extracellular matrix influence interactions.
The superficial dermis of adult human skin contains a complex arcading microvasculature that provides nutrient support to the overlying epidermis. We propose that the unique subpopulations of dermal fibroblasts located in the superficial dermis contribute to the organization and maintenance of this elaborate microvasculature. This possibility was tested in a coculture system in which distinct subpopulations of adult human dermal fibroblasts were grown to form high-density lawns that were then seeded with human umbilical vein vascular endothelial cells (EC). The fibroblast subpopulation cultured specifically from the papillary dermis supported a robust array of highly branched tube-like structures. In contrast, fibroblasts cultured from the reticular dermis provided an anemic level of support for the formation of tube-like structures. These varied interactions with vascular EC were not due to the differential production of the potent pro-angiogenic factors vascular endothelial growth factor-A or fibroblast growth factor-2. Instead, the extracellular matrix and/or molecules bound to this matrix appeared to contain instructions that modulated these differential fibroblast-vascular EC interactions. One matrix-binding growth factor, hepatocyte growth factor/scatter factor, was identified that was both differentially expressed by papillary and reticular dermal fibroblasts and which was shown to be physiologically relevant in the coculture model. These studies highlight the importance of fibroblasts in supporting and maintaining vascular integrity. Furthermore, these studies have important implications for wound repair and may help to explain how fibroblasts contribute to the etiology of nonhealing wounds.


The characteristics, management, and aftercare of patients with suicide attempts who attended the emergency department of a general hospital in northern Taiwan.
Suicide is a serious public health problem worldwide. The emergency department (ED) is often the first place of contact with medical and mental health care for suicidal patients. This study aimed to determine the characteristics, management, and aftercare of patients who attempted suicide and then were taken to the ED of a general hospital in Taipei, Taiwan. Our study was a cross-sectional retrospective study that consecutively recruited patients with suicide attempts attended to the ED of a general hospital from June 2004 to May 2005. The patients' medical information and records pertaining to their emergency visit were subsequently reviewed. Overall, 481 persons were recruited into our study. The male:female ratio of total surviving attempters was approximately 1:4 and the average age was 33.6 [standard deviation (SD) 12.3] years. The most frequently observed general characteristics from which participants suffered were mental illness (73.1%) and interpersonal problems (76.1%). Nearly all patients (91.7%) received ED on-site psychosocial assessments from social workers (SWs) and psychiatrists in 84.2% and 53.4% of cases, respectively. Less than half of patients (45.1%) were referred to psychiatric outpatient aftercare, and only 26.1% contacted the psychiatric outpatient clinics after discharge from the ED. The stated reasons for psychiatric outpatient referral were associated with interpersonal problems, current psychiatric illness, the ED on-site psychiatrist consultation, and admission to medical, surgical, or psychiatric wards. However, individuals with interpersonal problems, previous psychiatric intervention, and ED on-site psychiatrist consultation were significantly more likely to attend outpatient psychiatric aftercare. Individuals who harmed themselves had a high rate of psychiatric morbidity and interpersonal problems. However, their adherence to psychiatric outpatient aftercare was low. Improved identification of the needs of patients with suicidal tendencies who did not attend outpatient services will have implications for future services provided to this patient population, and will better enable medical personnel to most effectively assist in suicide attempt interventions.


IReNA: Integrated regulatory network analysis of single-cell transcriptomes and chromatin accessibility profiles.
Recently, single-cell RNA sequencing (scRNA-seq) and single-cell assay for transposase-accessible chromatin using sequencing (scATAC-seq) have been developed to separately measure transcriptomes and chromatin accessibility profiles at the single-cell resolution. However, few methods can reliably integrate these data to perform regulatory network analysis. Here, we developed integrated regulatory network analysis (IReNA) for network inference through the integrated analysis of scRNA-seq and scATAC-seq data, network modularization, transcription factor enrichment, and construction of simplified intermodular regulatory networks. Using public datasets, we showed that integrated network analysis of scRNA-seq data with scATAC-seq data is more precise to identify known regulators than scRNA-seq data analysis alone. Moreover, IReNA outperformed currently available methods in identifying known regulators. IReNA facilitates the systems-level understanding of biological regulatory mechanisms and is available at https://github.com/jiang-junyao/IReNA.


alpha 2B-Adrenoceptor levels govern agonist and inverse agonist responses in PC12 cells.
Receptor density is an important determinant of cellular effector responses to receptor activation. We analysed cytosolic Ca(2+) responses to alpha(2)-adrenergic agents in PC12 cells expressing human alpha(2B)-adrenergic receptors (AR) at two densities (3.8 and 1.3 pmol/mg protein). The efficacy (E(max)) of agonists was greater in cells with higher receptor expression; while the potency (EC(50)) of norepinephrine and oxymetazoline was independent of alpha(2B)-AR levels. Several classical alpha(2)-AR antagonists behaved as either partial or inverse agonists in a receptor density-dependent fashion. No apparent structural similarities were found among the inverse agonists, precluding simple predictions of inverse agonist activity. Transfected PC12 cells expressing alpha(2B)-AR at relatively high density would be a useful approach to screen inverse agonists for this class of receptors. Our results further indicate that receptor density significantly influences the properties of ligands, not only of partial agonists as predicted by classical receptor theory, but also of antagonists and full agonists.


A pilot radiometabolomics integration study for the characterization of renal oncocytic neoplasia.
Differentiating benign renal oncocytic tumors and malignant renal cell carcinoma (RCC) on imaging and histopathology is a critical problem that presents an everyday clinical challenge. This manuscript aims to demonstrate a novel methodology integrating metabolomics with radiomics features (RF) to differentiate between benign oncocytic neoplasia and malignant renal tumors. For this purpose, thirty-three renal tumors (14 renal oncocytic tumors and 19 RCC) were prospectively collected and histopathologically characterised. Matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) was used to extract metabolomics data, while RF were extracted from CT scans of the same tumors. Statistical integration was used to generate multilevel network communities of -omics features. Metabolites and RF critical for the differentiation between the two groups (delta centrality > 0.1) were used for pathway enrichment analysis and machine learning classifier (XGboost) development. Receiver operating characteristics (ROC) curves and areas under the curve (AUC) were used to assess classifier performance. Radiometabolomics analysis demonstrated differential network node configuration between benign and malignant renal tumors. Fourteen nodes (6 RF and 8 metabolites) were crucial in distinguishing between the two groups. The combined radiometabolomics model achieved an AUC of 86.4%, whereas metabolomics-only and radiomics-only classifiers achieved AUC of 72.7% and 68.2%, respectively. Analysis of significant metabolite nodes identified three distinct tumour clusters (malignant, benign, and mixed) and differentially enriched metabolic pathways. In conclusion, radiometabolomics integration has been presented as an approach to evaluate disease entities. In our case study, the method identified RF and metabolites important in differentiating between benign oncocytic neoplasia and malignant renal tumors, highlighting pathways differentially expressed between the two groups. Key metabolites and RF identified by radiometabolomics can be used to improve the identification and differentiation between renal neoplasms.


A tridimensional view of intradermal nevi as revealed by scanning electron microscopy.
When scanning electron microscopy (SEM) was applied to the study of intradermal nevi, interesting tridimensional features were recognized. The free surface of the lesions showed ruffled keratinized cells. "Normal" hairs as well as "corkscrew" hairs emerged from the follicular openings. Nevus cells were either round or elongated and surrounded by connective tissue fibers.


A highly active glutathione peroxidase in red blood cells of normal and acatalasemic mice.
The activities of catalase (E.C.184.108.40.206) and glutathione peroxidase (E.C.18.104.22.168) were compared in red blood cells from humans, ducks and normal and acatalasemic mice. In the cells from both strains of mice, an equally high activity of GSH-Px was found which could be inhibited completely by iodoacetate but was not sensitive to N-ethylmaleimide.


[Sarcoidosis of the kidney and skin without lung involvement].
We report the case of a patient with predominantly renal sarcoidosis. Renal failure responded well to systemic corticosteroid therapy. The clinical presentation was particular, notably the absence of lung involvement and the presence of cutaneous lymphedema.


Acute cadmium exposure induces stress-related gene expression in wild-type and metallothionein-I/II-null mice.
This study examined the effect of acute cadmium on stress-related gene expression and free radical production in wild-type and metallothionein-I/II-null (MT-null) mice. Atlas Toxicology arrays showed that acute cadmium (40 micromol/kg as CdCl(2), ip for 3 h) markedly increased the expression of genes encoding heat-shock proteins, heme oxygenase-1, and genes in response to DNA damage/repair. The expression of genes encoding cytochrome P450 enzymes, UDP-glucuronosyltransferases, Mn-superoxide dismutase, and catalase was suppressed by cadmium. MT-null mice were more sensitive than wild-type mice to cadmium-induced, stress-related gene expression, in accord with greater activation of transcription factor AP-1 and phosphorylated JNK and ERK. To evaluate free radical production, mice were simultaneously given the spin trap agent, N-tert-butyl-alpha-phenylnitrone (PBN, 250 mg in DMSO/kg, ip) with cadmium, and livers were removed 30 min later for PBN-trapped radical extraction with chloroform:methanol (2:1), and detected with electron spin resonance (ESR). Cadmium treatment caused detectable ESR signals for PBN adducts as well as lipid peroxidation in the liver similarly in both wild-type and MT-null mice. Thus, the mechanism of acute cadmium toxicity involves multiple facets including oxidative damage and aberrant gene expression, and absence of MT exacerbates Cd-induced aberrant gene expression.


Endoscopic management of adnexal masses.
The laparoscopic management of suspicious adnexal masses and early ovarian malignancies is discussed with the aim of maintaining accepted oncologic treatment principles. Comparative survival data of patients with gynecological malignancies managed by laparoscopy or laparotomy are still very scarce and the survival of cancer patients must not be compromised by new techniques. It is time to closely analyze laparoscopy and determine if it has a positive impact on the diagnosis and treatment of ovarian malignancies. In this paper we will address the following points: 1) Which ovarian cysts can be surgically treated by laparoscopy (pelviscopy)? 2) Is staging laparoscopy an accepted technique? 3) Is laparoscopy, as a second-look procedure, of benefit? 4) Is laparoscopic staging, together with histologic tissue sampling, adequate surgical technique in inoperable ovarian cancer with ascites and peritoneal carcinomatosis? 5) Does endoscopic biopsy of ovarian cancer stage Ia change the destiny of a patient into ovarian cancer Ic? The above questions are analyzed based on our experience with the laparoscopic treatment of 1,225 patients with ovarian cysts and 165 ovarian cancer patients stage I to IV treated immediately by laparotomy during the years 1992-1995. Ovarian cystic tumors with no signs of malignancy can be dealt with by laparoscopic means with the option of immediate conversion to laparotomy or within one week if an ovarian malignancy is diagnosed. Today sampling laparoscopic lymphadenectomy of both pelvic and para-aortic is feasible and adequate. On a curative level, the number of lymph nodes to be resected has yet to be determined. The adnexa can be extracted from the abdominal cavity with bag extraction without the danger of spillage. The uterus can be removed transvaginally with laparoscopic assisted vaginal hysterectomy (LAVH). We must be cautious to advocate laparoscopy for ovarian cancer. However, it is an excellent tool when used as a staging procedure. A careful preoperative screening of the patient and an exact definition of existing cysts with imaging techniques allows us to frequently apply laparoscopic surgery for ovarian cysts, leaving only readily detectable cancer cases for laparotomy. Many gynecological oncologists employing staging and second-look procedures for ovarian cancer agree that initiating a case with laparoscopy may preclude laparotomy for many patients. Tumor propagation by performing a biopsy in FIGO stage Ia ovarian cancer patients does not occur if the patient receives adequate radical surgical treatment within one week. According to the reports of Sevelda et al. and Dembo et al., the degree of differentiation and the existence of ascites are more relevant to decreasing the five-year survival rate of patients with ovarian cancer stage I than the rupture of capsule or penetration of the tumor. A dependency on the first two parameters was found in these two large statistical studies. As the question of endoscopic operations for adnexal mass is predominantly put for the sanitation of small ovarian tumors (ovarian tumors with solid particles in the cysts can be put into the section of primary laparotomies) there remains a wide field of indications for the laparoscopic treatment of adnexal mass and ovarian cysts with benign indications. For many young patients with non-malignant ovarian lesions such as endometriosis, benign cysts, benign cystic proliferations and fibromas, a laparotomy can be avoided and these lesions treated by laparoscopy.


Alteration of bcl-2 and bax expression in embolized meningiomas.
We examined the expression of the p53 and bcl-2 family in embolized meningiomas and investigated the interaction between them. p53 and its downstream effector p21 accumulated mainly in perinecrotic areas, where apoptosis was also observed. bcl-2 was often expressed in the areas distant from necrosis, whereas Bax was immunostained more intensely in the perinecrotic areas. Double staining for both p53 and Bcl-2 showed a reciprocal pattern. This appeared to be due to the down-regulation of Bcl-2 by p53. However, regulation of Bax by p53 was not distinct. The expression of p21 was not apparent 13 days after embolization, and apoptosis was observed until 6 days after embolization. The effect of embolization was most likely temporary, although growth arrest has been reported to persist for a long time in a limited number of embolized cases.


Interaction between inflammatory mediators and miRNAs in Helicobacter pylori infection.
Helicobacter pylori cause chronic inflammation favouring gastric carcinogenesis, and its eradication may prevent malignant transformation. We evaluated whether H. pylori infection and its eradication modify the expression of inflammatory mediators in patients with chronic gastritis. Furthermore, we assessed whether microRNAs modulate inflammatory pathways induced by H. pylori and identified miRNA-gene interaction networks. mRNA and protein expression of TNFA, IL6, IL1B, IL12A, IL2 and TGFBRII and miRNAs miR-103a-3p, miR-181c-5p, miR-370-3p, miR-375 and miR-223-3p were evaluated in tissue samples from 20 patients with chronic gastritis H. pylori negative (Hp-) and 31 H. pylori positive (Hp+), before and three months after bacterium eradication therapy, in comparison with a pool of Hp- normal gastric mucosa. Our results showed that H. pylori infection leads to up-regulation of TNFA, IL6, IL12A and IL2 and down-regulation of miRNAs. Bacterium eradication reduces the expression of TNFA and IL6 and up-regulates TGFBRII and all investigated miRNAs, except miR-223-3p. Moreover, transcriptional profiles of inflammatory mediators and miRNAs after eradication are different from the non-infected group. Deregulated miRNA-mRNA interaction networks were observed in the Hp+ group before and after eradication. Therefore, miRNAs modulated cytokine expression in the presence of H. pylori and after its eradication, suggesting that miRNAs participate in the pathological process triggered by H. pylori in the gastric mucosa.


Clinical significance of CTGF and Cry61 protein in extraocular muscles of strabismic patients.
To investigate the relationship between clinical features and protein amounts of Cysteine-rich 61 (Cyr61/CCN1) and connective tissue growth factor (CTGF/CCN2), which are vital components and regulators of the extracellular matrix in resected muscles from strabismus surgery. Strabismus patients who were diagnosed with horizontal concomitant strabismus or inferior oblique overaction (IOOA) and required extraocular muscles (EOMs) resection to correct eye position were included in this study. The protein amounts were measured by enzyme-linked immunosorbent assay (ELISA) in resected EOMs. Multivariable linear regression was used to investigate the associations, adjusting for gender, age (continuous), amblyopia, and disease duration. A total of 141 muscles (including 38 lateral, 81 medial rectus, and 22 inferior oblique muscles) from 128 patients were collected in this study. The amount of Cry61 and CTGF per millimeter was significantly negatively associated with deviation angle in intermittent exotropia patients (Cry61: β, - 1.44; 95%CI, - 2.79 to - 0.10, p = 0.035; CTGF: β, - 3.14; 95%CI, - 5.06 to - 1.22, p = 0.002). The same relationship was also detected in the partially accommodative and non-accommodative esotropia patients, although it was not statistically significant (Cry61: β, - 2.40; 95%CI, - 5.05 to 0.24; p = 0.073; CTGF: β, - 3.47; 95%CI, - 9.18 to 2.87; p = 0.269). The amount of Cry61 and CTGF per millimeter showed significant associations with the degree of IOOA (p < 0.05). Taken together, our results demonstrated a significant relationship between deviation angle and protein amount of Cry61 and CTGF and implied that Cry61 and CTGF may play important roles in modulation of EOM contractility, which provide new insights into strabismus pathogenesis.


Hemifusion arrest by complexin is relieved by Ca2+-synaptotagmin I.
Synaptic transmission relies on an exquisitely orchestrated series of protein-protein interactions. Here we show that fusion driven by neuronal SNAREs is inhibited by the regulatory protein complexin. Furthermore, inner-leaflet mixing is strongly impaired relative to total lipid mixing, indicating that inhibition by complexin arrests fusion at hemifusion. When the calcium sensor synaptotagmin is added in the presence of calcium, inhibition by complexin is relieved and full fusion rapidly proceeds.


Anti-fatigue effect of hypericin in a chronic forced exercise mouse model.
Hypericum perforatum L. is a traditional Chinese medicine used to sooth the liver, relieve depression, reduce body temperature, reduce sweating, and stimulate lactation. HP was extracted from Hypericum perforatum L. The antifatigue effects of hypericin were assessed in a series of experiments. Six-to eight-week-old male ICR mice were raised in our lab. Mice were subjected to swimming training for 2 h, 6 days/week for 6 weeks. One hour prior to each swimming session, intraperitoneal injection of saline or HP (2 or 4 mg/kg) was performed. Compared with the fatigue model control group, HP was found to significantly increase the swimming time in forced swimming tests. The molecular mechanisms underlying the antifatigue effects were further revealed by analysing energy metabolism, the oxidant-antioxidant system and the inflammatory response. HP normalized changes in BLA, LDH, BUN, and CK, LG in the liver. In addition, multiple assays have confirmed that HP improved the MDA, T-AOC, GSH-PX and SOD activity, and the relevant signalling pathways involved in the antifatigue effects were clarified. Furthermore, HP improves the expression of pro- and anti-inflammatory cytokines in skeletal muscle. These results suggested that the anti-chronic fatigue effects of HP are likely achieved by normalizing energy metabolism and attenuating oxidative and inflammatory responses. Consequently, this study supports HP use in the clinic to alleviate chronic fatigue.


Conserved and divergent features of neuronal CaMKII holoenzyme structure, function, and high-order assembly.
Neuronal CaMKII holoenzymes (α and β isoforms) enable molecular signal computation underlying learning and memory but also mediate excitotoxic neuronal death. Here, we provide a comparative analysis of these signaling devices, using single-particle electron microscopy (EM) in combination with biochemical and live-cell imaging studies. In the basal state, both isoforms assemble mainly as 12-mers (but also 14-mers and even 16-mers for the β isoform). CaMKIIα and β isoforms adopt an ensemble of extended activatable states (with average radius of 12.6 versus 16.8 nm, respectively), characterized by multiple transient intra- and inter-holoenzyme interactions associated with distinct functional properties. The extended state of CaMKIIβ allows direct resolution of intra-holoenzyme kinase domain dimers. These dimers could enable cooperative activation by calmodulin, which is observed for both isoforms. High-order CaMKII clustering mediated by inter-holoenzyme kinase domain dimerization is reduced for the β isoform for both basal and excitotoxicity-induced clusters, both in vitro and in neurons.


Association between short-term exposure to air pollutants and cause-specific daily mortality in Italy. A nationwide analysis.
Daily air pollution has been linked with mortality from urban studies. Associations in rural areas are still unclear and there is growing interest in testing the role that air pollution has on other causes of death. This study aims to evaluate the association between daily air pollution and cause-specific mortality in all 8092 Italian municipalities. Natural, cardiovascular, cardiac, ischemic, cerebrovascular, respiratory, metabolic, diabetes, nervous and psychiatric causes of death occurred in Italy were extracted during 2013-2015. Daily ambient PM10, PM2.5 and NO2 concentrations were estimated through machine learning algorithms. The associations between air pollutants and cause-specific mortality were estimated with a time-series approach using a two-stage analytic protocol where area-specific over-dispersed Poisson regression models where fit in the first stage, followed by a meta-analysis in the second. We tested for effect modification by sex, age class and the degree of urbanisation of the municipality. We estimated a positive association between PM10 and PM2.5 and the mortality from natural, cardiovascular, cardiac, respiratory and nervous system causes, but not with metabolic or psychiatric causes of death. In particular, mortality from nervous diseases increased by 4.55% (95% CI: 2.51-6.63) and 9.64% (95% CI: 5.76-13.65) for increments of 10 μg/m3 in PM10 and PM2.5 (lag 0-5 days), respectively. NO2 was positively associated with respiratory (6.68% (95% CI: 1.04-12.62)) and metabolic (7.30% (95% CI: 1.03-13.95)) mortality for increments of 10 μg/m3 (lag 0-5). Higher associations with natural mortality were found among the elderly, while there were no differential effects between sex or between rural and urban areas. Short-term exposure to particulate matter was associated with mortality from nervous diseases. Mortality from metabolic diseases was associated with NO2 exposure. Other associations are confirmed and updated, including the contribution of lowly urbanised areas. Health effects were also found in suburban and rural areas.


Association between parental behaviors and structural plasticity in the brain of male rodents.
In recent decades, human fathers across the globe have shown a substantial increase in their engagement in paternal caregiving behaviors. Despite the growing interest, the precise neurobiological mechanisms underlying caregiving behaviors in males remain unclear. Neurobiological studies conducted on rodents have advanced our understanding of the molecular, cellular, and circuit-level mechanisms. Typically, sexually naïve males exhibit aggression toward offspring, while fathers display parental behaviors. This drastic behavioral plasticity may be associated with changes in connections among specific regions or cell types. Recent studies have begun to describe this structural plasticity by comparing neural connections before and after fatherhood. In this Perspective, we summarize the findings from four well-studied rodent species, namely prairie voles, California mice, laboratory rats, and laboratory mice, with a view toward integrating past and current progress. We then review recent advances in the understanding of structural plasticity for parental behaviors. Finally, we discuss remaining questions that require further exploration to gain a deeper understanding of the neural mechanisms underlying paternal behaviors in males, including their possible implications for the human brain.


Potential of connected devices to optimize cattle reproduction.
Estrus and calving are two major events of reproduction that benefit from connected devices because of their crucial importance in herd economics and the amount of time required for their detection. The objectives of this review are to: 1) provide an update on performances reached by sensor systems to detect estrus and calving time; 2) discuss current economic issues related to connected devices for the management of cattle reproduction; 3) propose perspectives for these devices. The main physiological parameters monitored separately or in combination by connected devices are the cow activity, body temperature and rumination or eating behavior. The combination of several indicators in one sensor may maximize the performances of estrus and calving detection. An effort remains to be made for the prediction of calvings that will require human assistance (dystocia). The main reasons to invest in connected devices are to optimize herd reproductive performances and reduce labor on farm. The economic benefit was evaluated for estrus detection and depends on the initial herd performances, herd size, labor cost and price of the equipment. Major issues associated with the use of automated sensor systems are the weight of financial investment, the lack of economic analysis and limited skills of the users to manage associated technologies. In the near future, connected devices may allow a precise phenotyping of reproductive and health traits on animals and could help to improve animal welfare and public perception of animal production.


Cloning, expression and characterization of a putative 7alpha-hydroxysteroid dehydrogenase in Comamonas testosteroni.
The short-chain dehydrogenase/reductase (SDR) superfamily is a large and diverse group of genes with members found in all forms of life. Comamonas testosteroni (C. testosterone) ATCC11996 is a Gram-negative bacterium which can use steroids as carbon and energy source. In the present investigation, we found a novel SDR gene 7alpha-hydroxysteroid dehydrogenase (7α-HSD) which is located 11.9 kb upstream from hsdA with the same transcription orientation in the C. testosteroni genome. The open reading frame of this putative 7alpha-hydroxysteroid dehydrogenase gene consists of 771 bp and translates into a protein of 256 amino acids. Two consensus sequences of the SDR superfamily were found, an N-terminal Gly-X-X-X-Gly-X-Gly cofactor-binding motif and a Tyr-X-X-X-Lys segment (residues 161-165 in the 7α-HSD sequence) essential for catalytic activity of SDR proteins. To produce purified 7α-HSD protein, the 7α-HSD gene was cloned into plasmid p(ET-15b) and the over expressed protein was purified by His-tag sequence on metal chelate chromatography. To prove that 7α-HSD is involved in the metabolic pathway of steroid compounds, we constructed a 7α-HSD knock-out mutant of C. testosteroni. Compared to the wild type C. testosteroni, degradation of testosterone, estradiol and cholesterol were decreased in the 7α-HSD knock-out mutant. Furthermore, growth in the medium with testosterone, estradiol and cholesterol was impaired in 7α-HSD knock-out mutant. The results showed that 7α-HSD is involved in steroid degradation.


Association study of a (TG)n dinucleotide repeat at chromosome 15q13.3 and schizophrenia in the Chinese population.
Linkage studies have suggested that chromosome 15q13-q14 may harbor a susceptibility locus for schizophrenia. In the current study, the association between a (TG)n dinucleotide repeat polymorphism at D15S976 and schizophrenia was investigated using two independent samples from the Han Chinese population. In a population-based study, no significant difference was found between the genotype and allele frequency distributions in schizophrenia patients and control subjects. In a family-based study, no significant transmission disequilibrium from heterozygous parents to affected offspring was observed. Further analysis of the parent-of-origin effect found nominally significant allele-wise transmission disequilibrium through maternal transmissions, while 157bp and 159bp alleles showed significant individual allelic transmission disequilibrium from heterozygous mothers to affected offspring. Our results did not support the hypothesis that the (TG)n dinucleotide repeat polymorphism plays a major role in schizophrenia susceptibility in the Chinese population. Further studies are needed to elucidate the putative parent-of-origin effect and its role in schizophrenia susceptibility.


Relationship of dose to local control in advanced stage III and IV head and neck cancer treated by surgery and postoperative radiotherapy.
Retrospective analysis of 76 patients with locally advanced squamous cell carcinoma of multiple head and neck sites, who were treated by surgical resection and either 4,000-5,000, 6,000, or 6,600 rad postoperative external beam radiotherapy, revealed reduced local recurrence rates with increasing postoperative radiation dose.


Subfertility among HIV-affected couples in a safer conception cohort in South Africa.
Subfertility among couples affected by HIV has an impact on the well-being of couples who desire to have children and may prolong HIV exposure. Subfertility in the antiretroviral therapy era and its determinants have not yet been well characterized. The objective of the study was to investigate the burden and determinants of subfertility among HIV-affected couples seeking safer conception services in South Africa. Nonpregnant women and male partners in HIV seroconcordant or HIV discordant relationships desiring a child were enrolled in the Sakh'umndeni safer conception cohort at Witkoppen Clinic in Johannesburg between July 2013 and April 2017. Clients were followed up prospectively through pregnancy (if they conceived) or until 6 months of attempted conception, after which they were referred for infertility services. Subfertility was defined as not having conceived within 6 months of attempted conception. Robust Poisson regression was used to assess the association between baseline characteristics and subfertility outcomes; inverse probability weighting was used to account for missing data from women lost to safer conception care before 6 months of attempted conception. Among 334 couples enrolled, 65% experienced subfertility (inverse probability weighting weighted, 95% confidence interval, 0.59-0.73), of which 33% were primary subfertility and 67% secondary subfertility. Compared with HIV-negative women, HIV-positive women not on antiretroviral therapy had a 2-fold increased risk of subfertility (weighted and adjusted risk ratio, 2.00; 95% confidence interval, 1.19-3.34). Infertility risk was attenuated in women on antiretroviral therapy but remained elevated, even after ≥2 years on antiretroviral therapy (weighted and adjusted risk ratio, 1.63; 95% confidence interval, 0.98-2.69). Other factors associated with subfertility were female age (weighted and adjusted risk ratio, 1.03, 95% confidence interval, 1.01-1.05 per year), male HIV-positive status (weighted and adjusted risk ratio, 1.31; 95% confidence interval, 1.02-1.68), male smoking (weighted and adjusted risk ratio, 1.29; 95% confidence interval, 1.05-1.60), and trying to conceive for ≥1 year (weighted and adjusted risk ratio, 1.38; 95% confidence interval, 1.13-1.68). Two in 3 HIV-affected couples experienced subfertility. HIV-positive women were at increased risk of subfertility, even when on antiretroviral therapy. Both male and female HIV status were associated with subfertility. Subfertility is an underrecognized reproductive health problem in resource-limited settings and may contribute to prolonged HIV exposure and transmission within couples. Low-cost approaches for screening and treating subfertility in this population are needed.


A comparative study of the immunogold labeling on H(2)O(2)-treated and heated epoxy sections.
The purpose of this study was to compare the intensity of the immunogold labeling of H(2)O(2)-treated and heated epoxy sections. Renal swine tissue with glomerular immune complex deposits with reactivity against IgG was embedded in epoxy resin. Immunogold labeling with anti-IgG was performed on sections from these blocks. Some of these sections were treated by H(2)O(2), others were heated in a citrate solution, while some were not treated at all. Some epoxy sections, which had been exposed to both H(2)O(2) and heat, were also exposed to the same immunolabeling. The heated epoxy sections obtained an yield of specific immunogold labeling, which was twice as large as the labeling of the H(2)O(2)-treated sections. The yield of immunolabeling of the sections that had been exposed to both H(2)O(2) and heat was not significantly different from the sections that were only exposed to heat. The non-treated sections were very weakly labeled with anti-IgG. We believe that both H(2)O(2) and heat have the ability to break some chemical bonds between the epoxy resin and the antigens, but heating in citrate buffer has a larger potential in this respect than H(2)O(2). We interpret the results from the combined treatment with H(2)O(2) and heat in the following way; the bonds that are broken by H(2)O(2) will also be broken by heating in citrate solution. The practical significance of these results is that heating in citrate buffer is a more convenient method for enhancing the immunolabeling of epoxy sections than treatment with H(2)O(2).


Farm riparian land use and management: driving factors and tensions between technical and ecological functions.
Because of their specific role in the fate of natural resources of landscapes, riparian lands are considered as landscape management units. Yet, they are part of many farms: In what way does this influence their land use and sustainability? In this article, farm riparian land was defined as the set of riparian fields of a given farm. Our aim was to evaluate farm riparian lands as farm management units and the balance between technical and ecological functions associated with farm riparian land-use types. Technical functions designated the role of land use in elaborating farm production and maintaining farm territory. Ecological functions corresponded to farm riparian land-use intensity and ratio of permanent vegetation. The analysis was carried on 102 farms from 5 study sites representative of dairy agriculture and landscapes in a region straddling Brittany and Normandy (France). Farm riparian land-use types were identified using statistical clustering. Technical and ecological functions associated with land-use types were expressed in the light of agronomy and landscape ecology expertise. Descriptors of farm holdings and farm riparian land were tested as explanatory factors of farm riparian land-use types. The use of farm riparian lands was diverse but well defined; they proved to be farm management units. Compatibilities or antagonisms between technical and ecological functions were underscored according to farm riparian land-use type. We argued that decision support could gain in combining perspectives on riparian lands as landscape management units and farm management units.


Preserving protein function through reversible aggregation.
It is generally accepted that protein function depends on a defined 3D structure, with unfolding and aggregation dealing a final blow to functionality. A study now shows that the regulated exposure of an unstructured region in yeast pyruvate kinase triggers reversible aggregation to preserve protein function under stress.


Synthesis and biological evaluation of Esaprazole analogues showing σ1 binding and neuroprotective properties in vitro.
Esaprazole, a molecule previously acknowledged to protect against stomach and intestinal ulcers was surprisingly discovered to have neuroprotective activities and σ1 binding in vitro. A highly diverse set of Esaprazole analogues 2-5 was prepared in order to increase blood-brain barrier penetration. The analogues showed a structure-activity relationship at the σ1 receptor closely matching already published pharmacophores. Many of the analogues were shown to have neuroprotective properties in two assays using primary cultures of cortical neurons exposed to glutamate and hydrogen peroxide. However, no apparent SAR for these two assays could be developed. Metabolic stability of the analogues were also investigated and the structure of R(1) had a significant bearing on the ADME properties of the compound resulting in two series of compounds. Compounds in which R(1) was a H or acyl group had good metabolic stability in RLM but poor BBB penetration, whereas compounds where R(1) was a cyclo- or bicyclo-alkyl group had poor metabolic stability but good BBB penetration.


Design of a functional cyclic HSV1-TK reporter and its application to PET imaging of apoptosis.
Positron emission tomography (PET) is a sensitive and noninvasive imaging method that is widely used to explore molecular events in living subjects. PET can precisely and quantitatively evaluate cellular apoptosis, which has a crucial role in various physiological and pathological processes. In this protocol, we describe the design and use of an engineered cyclic herpes simplex virus 1-thymidine kinase (HSV1-TK) PET reporter whose kinase activity is specifically switched on by apoptosis. The expression of cyclic TK (cTK) in healthy cells leads to inactive product, whereas the activation of apoptosis through the caspase-3 pathway cleaves cTK, thus restoring its activity and enabling PET imaging. In addition to detailing the design and construction of the cTK plasmid in this protocol, we include assays for evaluating the function and specificity of the cTK reporter in apoptotic cells, such as assays for measuring the cell uptake of PET tracer in apoptotic cells, correlating doxorubicin (Dox)-induced cell apoptosis to cTK function recovery, and in vivo PET imaging of cancer cell apoptosis, and we also include corresponding data acquisition methods. The time to build the entire cTK reporter is ∼2-3 weeks. The selection of a stable cancer cell line takes ∼4-6 weeks. The time to implement assays regarding cTK function in apoptotic cells and the in vivo imaging varies depending on the experiment. The cyclization strategy described in this protocol can also be adapted to create other reporter systems for broad biomedical applications.


A liver-specific microRNA binds to a highly conserved RNA sequence of hepatitis B virus and negatively regulates viral gene expression and replication.
Regulated gene expression and progeny production are essential for persistent and chronic infection by human pathogens, such as hepatitis B virus (HBV), which affects >400 million people worldwide and is a major cause of liver disease. In this study, we provide the first direct evidence that a liver-specific microRNA, miR-122, binds to a highly conserved HBV pregenomic RNA sequence via base-pairing interactions and inhibits HBV gene expression and replication. The miR-122 target sequence is located at the coding region of the mRNA for the viral polymerase and the 3' untranslated region of the mRNA for the core protein. In cultured cells, HBV gene expression and replication reduces with increased expression of miR-122, and the expression of miR-122 decreases in the presence of HBV infection and replication. Furthermore, analyses of clinical samples demonstrated an inverse linear correlation in vivo between the miR-122 level and the viral loads in the peripheral blood mononuclear cells of HBV-positive patients. Our results suggest that miR-122 may down-regulate HBV replication by binding to the viral target sequence, contributing to the persistent/chronic infection of HBV, and that HBV-induced modulation of miR-122 expression may represent a mechanism to facilitate viral pathogenesis.


Pathophysiology of lipoprotein transport.
A system for classification of genetic and acquired forms of hyperlipidemia in humans based on lipoprotein physiology is described. Most hyperlipidemia can be accounted for by defects in one of four sites of physiologic regulation: (1) triglyceride-rich lipoprotein production, (2) lipoprotein lipase-mediated triglyceride catabolism, (3) remnant lipoprotein catabolism, and (4) extrahepatic cholesterol-rich lipoprotein catabolism.


Identification on I-Ak molecules of a functional site recognized by proliferating T-lymphocytes.
The inhibitory capacity of 17 monoclonal antibodies (m.Ab.) specific for the products of the I-Ak subregion was evaluated in proliferative responses of B10.BR T-lymphocytes to GAT, Keyhole limpet hemocyanin, and ovalbumin. Considered in isolation, each m.Ab. mediated inhibitory effects of comparable magnitude on these three different proliferative responses. On the other hand, clear differences were observed when the magnitude of the inhibitory effects was compared from one m.Ab. to another. The m.Ab. were consequently classified as strong or moderate-to-weak inhibitors of T-cell proliferative responses. Evidence was simultaneously gained indicating the following: (a) the determinants recognized by different m.Ab. were expressed on the same molecules; (b) the differences in affinity of the m.Ab. for I-Ak positive cells did not explain their differences in inhibitory capacities; (c) conversely, the inhibitory capacity of each m.Ab. followed its ability to inhibit the cell surface fixation of Ia.17-specific 10-2.16 m.Ab; (d) the strong inhibitory capacity of some m.Ab. was not related to a special ability to modulate cell surface Ia molecules. These results suggest that antigen recognition by T lymphocytes is preferentially restricted by a functional site of the I-Ak molecules related to the Ia.17 and Ia.1 specificities.


Correction to: KIF5B-RET fusion kinase promotes cell growth by multilevel activation of STAT3 in lung cancer.
After the publication of this work [1].


Hepatic triglyceride lipase is not an insulin-dependent enzyme in rats.
The aim of this study is to examine whether insulin regulates the activity of hepatic triglyceride lipase. The activities of both hepatic triglyceride lipase (H-TGL) and lipoprotein lipase (LPL) were measured in plasma after intravenous administration of heparin. Insulin-deficient diabetic rats had normal H-TGL activity. Insulin-treated normal rats had no different enzyme activities from those of controls. Ventromedial hypothalamic (VMH) lesioned rats that became obese and showed hyperinsulinemia, had also normal enzyme activities. From these experiments, it is concluded that H-TGL is not an insulin-dependent enzyme. As far as LPL is concerned, this enzyme was increased in both insulin-treated rats and rats with VMH lesions. This was consistent with the established fact that insulin has a stimulatory effect on adipose tissue LPL. The LPL activity in postheparin plasma was not decreased in insulin-deficient rats. It is speculated that a possible increase in muscle LPL compensated for the low adipose tissue LPL in diabetic rats.


Combined radiation-protective and radiation-sensitizing agents. IV: Measurement of intracellular protector concentrations.
Radiosensitization of hypoxic V79 Chinese hamster cells by 0.5 mM misonidazole at approximately 0-4 degrees C is substantially enhanced by pretreating the cells overnight with 0.1 mM buthionine sulfoximine, which lowers the cellular glutathione content to 5% of control values (from 4 mM to approximately 0.2 mM). The enhanced sensitization is reversed by concentrations of exogenous cysteine that are much lower (0.02 mM) than the original glutathione content. Reduced Co-enzyme A affords reversal of the enhancing effect at concentrations of about 1 mM. Sodium ascorbate gives no protection at all even at concentrations of 2 mM. The intracellular concentration of the reducing agents was measured using a spin-through oil technique. There was no diffusion of Co-A (MW greater than 750) or ascorbate (excluded by charge) into the cells. In contrast, cysteine was rapidly concentrated by factors of 4-10, even at the low temperatures used. Extracellular ascorbate's inability to radioprotect argues against electron transfer across the cell membrane as a mechanism for radioprotection. This mechanism could have explained the ability of exogenous thiols to radioprotect in former studies using glutathione, and in the present studies using Co-A. The potential of cysteine to be concentrated by cells poses a problem in the interpretation of "exogenous protection" by non-diffusing thiols, since trace contamination by cysteine could lead to the actual protection observed. Cysteine could also be formed by exchange reactions of exogenous thiols with the disulfide of cysteine, present in all media formulations.


Two new nor-lignans from Selaginella pulvinata (Hook. & Grev.) Maxim and their antihyperglycemic activities.
Two new nor-lignans, pulvin A (1) and moellenoside C (2), along with two known compounds (3-4) were isolated from the whole plant of Selaginella pulvinate (Hook. & Grev.) Maxim. The structures of the new compounds were established on the basis of spectroscopic data and acid hydrolysis. All the isolates were investigated for their antihyperglycemic activities in 3T3-L1 adipocytes. The results showed that compounds 1 and 2 promoted the glucose consumption prominently in 3T3-L1 adipocytes in a dose-response manner. Compound 1 and 2 induced 1.14-1.73 folds and 1.03-1.55 folds changes relative to the basal level, respectively, in the concentration range of 12.5 μM to 50 μM.


A quantitative morphological study of HRP-labelled cat alpha-motoneurones supplying different hindlimb muscles.
Cat alpha-motoneurones supplying the quadriceps (Q), posterior biceps (PB), gastrocnemius (G), soleus (SOL) and short intrinsic plantar foot (SP) muscles were studied after retrograde or intracellular labelling with HRP. The average soma sizes were rather similar for the different pools, the SOL cells being the smallest. The median number of first-order dendrites ranged from 10 (PB) to 12 (SOL). The median diameters of the first-order dendrites ranged from 6 (SOL) to 8.5 (PB, G) micrometer. The dendritic projection patterns were rather similar for the different motoneurone groups, except for a prominent dorsomedial projection of SP dendrites. A considerable fraction of the dendrites extended into the white matter. The diameter of the first-order dendrite correlated positively to the number of end branches as well as to the combined length, surface area and volume of the whole dendrite. These relations appeared to be independent of motoneurone group and dendritic orientation. The combined diameter of the first-order dendrites, which reflects the total dendritic size of a motoneurone, exhibited median values between 82 micrometers (SOL) and 112 micrometers (Q). With respect to the relative scaling of soma and dendrites, motoneurones with large somas tended to have proportionally larger dendritic trees. The distribution of dendritic diameters, number of branches, dendritic surface area and volume, and the combined dendritic parameter (epsilon d3/2) at various distances from the soma were quite similar for the different motoneurone groups.


Two-stage banana leaves wastes utilization towards mushroom growth and biogas production.
Banana leaves wastes (BL) were subjected to fungal treatment using Pleurotus ostreatus to produce edible mushrooms and biogas in the anaerobic digestion process. Effects of fungal treatment on mushrooms production, lignin degradation, trace elements compositions and biogas yield during the anaerobic digestion process were evaluated. Treatment with P.ostreatus for 36 d resulted in the production of 181 ± 19 g of edible mushrooms per 2 kg of BL with biological efficiency of 37 ± 4%. Lignin concentration in fungal treated BL decreased by 10% indicating an improvement on its digestibility. Important trace elements (Fe, Mn, Mo, Co and Ni) necessary for the improvement of the anaerobic digestion process were also significantly reduced (P < 0.05) during the fungal treatment process. The biogas yield for the fungal treated BL was 282 mL g-1 VS-1 of which this study suggests that could be improved through trace element supplementation during the anaerobic digestion process.


Effective intervals for the administration of estradiol pulses and the induction of sexual behavior in female rats.
The interval between two pulses of estradiol was varied and subsequent progesterone-facilitated female sexual behavior was observed. The total dose of estradiol (2 micrograms free estradiol) has been shown previously to elicit little receptivity when administered in a single pulse. In the present study, two pulses of 1.0 microgram free estradiol separated by anywhere from 3-36 hours induced high levels of progesterone-facilitated receptivity when observed 18 hours after the second estrogen pulse. Various intervals between the second estrogen pulse and the time of testing were also evaluated. Female rats tested at 18, 24 and 36 hours after the second estrogen pulse (plus progesterone) were highly receptive. These results raise new possibilities about the time course of cellular events underlying the estrogen induction of feminine sexual behavior.


Changes in cerebral blood volume and amyloid pathology in aged Alzheimer APP/PS1 mice on a docosahexaenoic acid (DHA) diet or cholesterol enriched Typical Western Diet (TWD).
High dietary cholesterol and low dietary docosahexaenoic acid (DHA) intake are risk factors for Alzheimer's disease (AD). However, it is unclear how these components influence the course of the disease. We investigated the effects of dietary lipids on beta-amyloid deposition and blood circulation in the brains of 18-month-old APP/PS1 mice. Starting at 6 months of age, mice were fed a regular rodent chow, a Typical Western Diet (TWD) containing 1% cholesterol, or a diet with a high (0.5%) level of DHA for 12 months. Relative cerebral blood volume (rCBV) and flow (CBF) were determined with (2)H MR spectroscopy and gradient echo contrast enhanced MRI. Deposition of beta-amyloid was visualized in fixed brain tissue with immunohistochemistry. The TWD diet increased plaque burden in the dentate gyrus of the hippocampus, but did not significantly reduce rCBV. In contrast, the DHA-enriched diet increased rCBV without changing blood flow indicating a larger circulation in the brain probably due to vasodilatation and decreased the amount of vascular beta-amyloid deposition. Together, our results indicate that the long-term intake of dietary lipids can impact both brain circulation and beta-amyloid deposition, and support the involvement of hemodynamic changes in the development of AD.


Lysosomal enzymes in experimental allergic encephalomyelitis: time course and evidence of the source.
The lysosomal enzymes acid proteinase and beta-glucuronidase, were assayed in spinal cords of rats during the course of experimental allergic encephalomyelitis (EAE). Histological and histochemical examination was carried out versus controls, in selected areas of the same cords biochemically assayed, to look at the distribution of the lysosomal enzyme acid phosphatase. The biochemical assay showed a significant increase of the enzyme activities during the disease and the increase was significantly correlated with the intensity of the disease. The distribution in the nervous tissue of the increase in acid phosphatase activity observed in animals with EAE, suggests that endogenous nervous cells may contribute to the lysosomal enzyme increase in EAE.


[A scintigraphic simulated cold nodule caused by a cranial course of the subclavian artery and vein].
A patient who underwent a diagnostic thyroid scan, showed an anechoic area in the right caudal lobe by ultrasound. In the thyroid scan this area appeared as a cold spot. Coloured duplex ultrasound revealed that this area corresponded to the subclavian artery and vein and thus represented a deviation from the normal anatomy of these vessels.