[The heart during pregnancy].
During pregnancy, there are a number of important changes to cardiovascular function which are necessary for progression of a successful pregnancy. Additionally, preexisting cardiovascular conditions can be exacerbated by the adaptations that occur during gestation. These can present serious therapeutic challenges in the management of the cardiology patient during pregnancy. Significantly, the number of pregnant women at risk of cardiovascular complications is on the rise, so identification of risk factors that predict cardiac outcomes is essential to proper screening of the obstetrical patient. In diagnosed preexisting conditions, such as pulmonary hypertension, counseling is important prior to pregnancy. In the case of underlying disorders unmasked by pregnancy, or new-onset complications like preeclampsia, appropriate monitoring and treatment of the cardiovascular complications is warranted. Ultimately, collaborative care by both obstetricians and cardiologists is essential for the successful resolution of cardiovascular dysfunction in the obstetrical patient.


When bigger is better: the role of polyploidy in organogenesis.
Defining how organ size is regulated, a process controlled not only by the number of cells but also by the size of the cells, is a frontier in developmental biology. Large cells are produced by increasing DNA content or ploidy, a developmental strategy employed throughout the plant and animal kingdoms. The widespread use of polyploidy during cell differentiation makes it important to define how this hypertrophy contributes to organogenesis. I discuss here examples from a variety of animals and plants in which polyploidy controls organ size, the size and function of specific tissues within an organ, or the differentiated properties of cells. In addition, I highlight how polyploidy functions in wound healing and tissue regeneration.


Highly sensitive microfluidic competitive enzyme immunoassay based on chemiluminescence resonance energy transfer for the detection of neuron-specific enolase.
A microfluidic competitive enzyme immunoassay based on chemiluminescence resonance energy transfer (CRET) was developed for highly sensitive detection of neuron-specific enolase (NSE). The CRET system consisted of horseradish peroxidase (HRP)/luminol as a light donor and fluorescein isothiocyanate as an acceptor. When fluorescein isothiocyanate-labeled antibody binds with HRP-labeled antigen to form immunocomplex, the donor and acceptor are brought close each other and CRET occurs in the immunocomplex. In the MCE, the immunocomplex and excess HRP-NSE were separated, and the chemiluminescense intensity of immunocomplex was used to estimate NSE concentration. The calibration curve showed a linearity in the range of NSE concentrations from 9.0 to 950 pM with a correlation coefficient of 0.9964. Based on a S/N of 3, the detection limit for NSE determination was estimated to be 4.5 pM, which is two-order magnitude lower than that of without CRET detection. This assay was applied for NSE quantification in human serum. The obtained results demonstrated that the proposed immunoassay may serve as an alternative tool for clinical analysis of NSE.


First in vivo assessment of RAS-Q technology as lung support device for pulmonary hypertension.
To assess the in vivo hemodynamic effects on the pressure overloaded right ventricle of RAS-Q® technology, the world's first gas exchanger with a fully integrated compliance. In six acute in vivo trials RAS-Q was implanted in sheep between the pulmonary artery and left atrium. Right ventricular pressure overload was induced by pulmonary artery banding. Pressures and flows were recorded in baseline, moderate and severe pulmonary hypertension conditions. In one trial, RAS-Q was benchmarked against the pediatric Quadrox-i®. With 1.00 and 1.17 L/min, RAS-Q delivered 31% and 39% of the total cardiac output in moderate and severe pulmonary hypertension, respectively. Pulmonary artery pressures and mean pulmonary artery pressure/mean arterial blood pressure ratio successfully decreased, implying a successful right ventricular unloading. Cardiac output was restored to normal levels in both pulmonary hypertension conditions. With both devices in parallel, RAS-Q provided three times higher flow rates and a 10 times higher pressure relief, compared to the pediatric Quadrox-i. A gas exchanger with a fully integrated compliance better unloads the right ventricle compared to a non-compliant gas exchanger and it can restore cardiac output to normal levels in cases of severe pulmonary hypertension.


Biomechanical assessment of remote and postinfarction scar remodeling following myocardial infarction.
The importance of collagen remodeling following myocardial infarction (MI) is extensively investigated, but little is known on the biomechanical impact of fibrillar collagen on left ventricle post-MI. We aim to identify the significant effects of the biomechanics of types I, III, and V collagen on physio-pathological changes of murine hearts leading to heart failure. Immediately post-MI, heart reduces its function (EF = 40.94 ± 2.12%) while sarcomeres' dimensions are unchanged. Strikingly, as determined by immunohistochemistry staining, type V collagen fraction significantly grows in remote and scar for sustaining de novo-types I and III collagen fibers' assembly while hindering their enzymatic degradation. Thereafter, the compensatory heart function (EF = 63.04 ± 3.16%) associates with steady development of types I and III collagen in a stiff remote (12.79 ± 1.09 MPa) and scar (22.40 ± 1.08 MPa). In remote, the soft de novo-type III collagen uncoils preventing further expansion of elongated sarcomeres (2.7 ± 0.3 mm). Once the compensatory mechanisms are surpassed, the increased turnover of stiff type I collagen (>50%) lead to a pseudo-stable biomechanical regime of the heart (≅9 MPa) with reduced EF (50.55 ± 3.25%). These end-characteristics represent the common scenario evidenced in patients suffering from heart failure after MI. Our pre-clinical data advances the understanding of the cause of heart failure induced in patients with extended MI.


Progranulin Improves Acute Lung Injury through Regulating the Differentiation of Regulatory T Cells and Interleukin-10 Immunomodulation to Promote Macrophage Polarization.
Progranulin (PGRN), which plays an anti-inflammatory role in acute lung injury (ALI), is promising as a potential drug. Studies have shown that regulatory T cells (Tregs) and interleukin- (IL-) 10 can repress inflammation and alleviate tissue damage during ALI. In this study, we built a lipopolysaccharide- (LPS-) induced ALI mouse model to illustrate the effect of PGRN on regulation of Treg differentiation and modulation of IL-10 promoting macrophage polarization. We found that the proportion of Tregs in splenic mononuclear cells and peripheral blood mononuclear cells was higher after treatment with PGRN. The increased proportion of Tregs after PGRN intratracheal instillation was consistent with the decreased severity of lung injury, the reduction of proinflammatory cytokines, and the increase of anti-inflammatory cytokines. In vitro, the percentages of CD4+CD25+FOXP3+ Tregs from splenic naïve CD4+ T cells increased after PGRN treatment. In further research, it was found that PGRN can regulate the anti-inflammatory factor IL-10 and affect the polarization of M1/M2 macrophages by upregulating IL-10. These findings show that PGRN likely plays a protective role in ALI by promoting Treg differentiation and activating IL-10 immunomodulation.


The fifth Datta Lecture. Structural similarities between the aspartate receptor of bacterial chemotaxis and the trp repressor of E. coli. Implications for transmembrane signaling.
A high resolution structure of the N-terminal ligand-binding domain of the aspartate receptor which mediates aspartate chemotaxis in Salmonella typhimurium has recently been reported. A least-squares superposition of the alpha-amino nitrogen, alpha-carbon, beta-carbon, and alpha-carboxylate carbon of the aspartate bound to the aspartate receptor onto the equivalent atoms in the tryptophan bound to the trp repressor provides evidence for similarity between key parts of the active sites that bind to the alpha-amino and alpha-carboxylates of the respective ligands. Because the N-terminal domain of the aspartate receptor and the trp repressor also share other structural similarities, we hypothesize that the similarity between the aspartate receptor and the trp repressor derives from a similarity in ligand-induced conformational changes at the active sites of these proteins. This hypothesis also implies that an important signaling event in the aspartate receptor occurs through tertiary conformational changes within a single subunit.


Pair formation in a coral inhabiting hermit crab.
The hermit crab, Paguritta harmsi, is a sedentary filterfeeder, previously known as an inhabitant of polychaete tubes on the surface of living corals. It is now reported living directly within colonies of the massive coral, Astreopora myriopthalma, growing in 8-10 m of water at Heron Island, Great Barrier Reef, Australia. Small hermit crabs (2-3 mm body length) were found within normal, tissue-containing corallites of the host, while larger individuals occupied the outer portion of tubular pits that extended as much as 98 mm down into the coral skeleton. P. harmsi tended to occur in pairs of adjacent male and female individuals, which may reduce predation risks during mating. Available evidence favours the hypothesis that hermit crabs are quite long lived, invade normal host corallites and remain there while the coral gradually grows outward producing an elongate pit.


The Relationships between Polymorphisms in Genes Encoding the Growth Factors TGF-β1, PDGFB, EGF, bFGF and VEGF-A and the Restenosis Process in Patients with Stable Coronary Artery Disease Treated with Bare Metal Stent.
Neointima forming after stent implantation consists of vascular smooth muscle cells (VSMCs) in 90%. Growth factors TGF-β1, PDGFB, EGF, bFGF and VEGF-A play an important role in VSMC proliferation and migration to the tunica intima after arterial wall injury. The aim of this paper was an analysis of functional polymorphisms in genes encoding TGF-β1, PDGFB, EGF, bFGF and VEGF-A in relation to in-stent restenosis (ISR). 265 patients with a stable coronary artery disease (SCAD) hospitalized in our center in the years 2007-2011 were included in the study. All patients underwent stent implantation at admission to the hospital and had another coronary angiography performed due to recurrence of the ailments or a positive result of the test assessing the coronary flow reserve. Angiographically significant ISR was defined as stenosis >50% in the stented coronary artery segment. The patients were divided into two groups-with angiographically significant ISR (n = 53) and without significant ISR (n = 212). Additionally, the assessment of late lumen loss (LLL) in vessel was performed. EGF rs4444903 polymorphism was genotyped using the PCR-RFLP method whilst rs1800470 (TGFB1), rs2285094 (PDGFB) rs308395 (bFGF) and rs699947 (VEGF-A) were determined using the TaqMan method. Angiographically significant ISR was significantly less frequently observed in the group of patients with the A/A genotype of rs1800470 polymorphism (TGFB1) versus patients with A/G and G/G genotypes. In the multivariable analysis, LLL was significantly lower in patients with the A/A genotype of rs1800470 (TGFB1) versus those with the A/G and G/G genotypes and higher in patients with the A/A genotype of the VEGF-A polymorphism versus the A/C and C/C genotypes. The C/C genotype of rs2285094 (PDGFB) was associated with greater LLL compared to C/T heterozygotes and T/T homozygotes. The polymorphisms rs1800470, rs2285094 and rs6999447 of the TGFB1, PDGFB and VEGF-A genes, respectively, are associated with LLL in patients with SCAD treated by PCI with a metal stent implantation.


Safety and efficacy of colesevelam hydrochloride in combination with fenofibrate for the treatment of mixed hyperlipidemia.
The aim of this study was to evaluate the amount of low-density lipoprotein cholesterol (LDL-C) reduction achieved by adding the specifically engineered bile acid sequestrant (SE-BAS) colesevelam HCl to a stable dose of fenofibrate in patients with mixed hyperlipidemia. Patients with mixed hyperlipidemia (n = 129) were enrolled in a randomized, double-blind, placebo-controlled, parallel-group study investigating the efficacy of fenofibrate plus colesevelam HCl versus fenofibrate monotherapy. After a 4- to 8-week washout period, subjects received fenofibrate 160 mg/day for 8 weeks and were then randomized to receive colesevelam HCl 3.75 g/day or placebo, in addition to fenofibrate 160 mg/day, for 6 weeks. The primary efficacy endpoint was mean percent change in LDL-C during randomized treatment. Secondary endpoints included absolute and percent changes in mean levels of LDL-C, triglycerides (TGs), high-density lipoprotein cholesterol (HDL-C), non-HDL-C, total cholesterol (TC), and apolipoproteins (apo) A-I and B during randomized treatment and from washout to end of randomized treatment. Of the 129 patients randomized to treatment, 119 completed the study. After 6 weeks of treatment, fenofibrate plus colesevelam HCl produced a mean percent change in LDL-C of -10.4% versus +2.3% with fenofibrate monotherapy (p < 0.0001). Fenofibrate plus colesevelam HCl was significantly more effective than fenofibrate alone at reducing levels of non-HDL-C, TC, and apo B (p < or = 0.0002). Colesevelam HCl did not significantly affect the TG-lowering effects of fenofibrate. Both treatment regimens were safe and well tolerated. Compared with fenofibrate monotherapy in patients with mixed hyperlipidemia, fenofibrate/colesevelam HCl combination therapy significantly reduced mean LDL-C, non-HDL-C, TC, and apo B levels without significantly affecting the TG-lowering or HDL-C-raising effects of fenofibrate. Fenofibrate/colesevelam HCl combination therapy is a safe, useful alternative for the treatment of mixed hyperlipidemia.


Thyroid hormone regulates the mouse thyrotropin beta-subunit gene promoter in transfected primary thyrotropes.
In TtT 97 cells, a thyrotropin-producing mouse pituitary tumor, thyroid hormone rapidly inhibits the transcription rate of both the thyrotropin alpha- and beta-subunit (TSH beta) genes, and this closely parallels the increase in nuclear thyroid hormone receptor occupancy. In this study, we have identified regions of the mouse TSH beta gene which are involved in mediating tissue-specific and thyroid hormone-regulated expression. Transient expression studies were performed using a series of chimeric plasmids in which 5'-flanking DNA was ligated to the firefly luciferase gene. Following transfection by electroporation, efficient expression of TSH beta 5'-flanking luciferase constructs occurred only in cells derived from TtT 97 tumors which express the endogenous TSH beta gene. Deletion analysis demonstrated that the region of the 5'-flanking DNA between positions -271 and -80 relative to the major transcriptional start site is important for TSH beta promoter activity in thyrotropes. No expression was measurable in mouse L cells, a fibroblast line, whereas a low level of expression was seen in MGH 101A cells derived from a thyrotropic tumor which no longer expresses the TSH beta gene. Reduced expression of TSH beta constructs was also found in GH3 and GH4 pituitary tumor lines. Addition of thyroid hormone effectively inhibited the level of transient TSH beta promoter activity in TtT 97 cells in a dose-dependent manner. The inhibitory effect was more pronounced and more accurately reflected the transcription rate data when transfected cells were derived from tumors treated with thyroid hormone for 5 days prior to transfection. Deletion of all but 46 base pairs of TSH beta gene 5'-flanking DNA and 3 base pairs of the first exon had no effect on thyroid hormone inhibition. This indicates that signals sufficient for transcriptional regulation of the TSH beta gene by thyroid hormone reside in the vicinity of the proximal promoter and may act by interfering with basal transcriptional factors.


Abnormal relaxation rates in subjects susceptible to malignant hyperthermia.
The acute malignant hyperthermia crisis appears to be mediated by a sudden and sustained rise in the concentration of sarcoplasmic calcium, leading to a prolonged muscle contraction. It is not known whether calcium metabolism in skeletal muscle is abnormal in malignant hyperthermia susceptible (MHS) subjects under normal conditions. A hypermetabolic state in skeletal muscle can be assessed in vivo by measuring relaxation rate. In the present study the relaxation rate was measured in the adductor pollicis muscle after electrical stimulation of the ulnar nerve. Ten MHS subjects with no clinical symptoms of muscle disease were investigated. Relaxation rates were found to be significantly higher in the MH-susceptible subjects compared with controls even under normal conditions (p less than 0.001). Determination of relaxation rate could thus be considered a possible addition to those methods currently available for the diagnosis of MHS.


Prevalence of premenstrual syndrome and premenstrual dysphoric disorder in Japanese high school students.
To determine the prevalence and the impact of premenstrual symptoms among Japanese adolescent girls, a total of 618 high school students were assessed. Of them, 64.6% were found to suffer from premenstrual symptoms, which is lower than that in adult women. On the other hand, the rates of prevalence of moderate to severe PMS and PMDD in girls were higher than those in adult women. Premenstrual symptoms could have significant consequences by interfering with the daily functioning of adolescent girls.


Hepatic artery reconstruction using an operating microscope in pediatric liver transplantation-Is it worth the effort?
In pediatric liver transplantation (pLT), hepatic artery thrombosis (HAT) is associated with inferior transplant outcome. Hepatic artery reconstruction (HAR) using an operating microscope (OM) is considered to reduce the incidence of HAT. HAR using an OM was compared to a historic cohort using surgical loupes (SL) in pLT performed between 2009 and 2020. Primary endpoint was the occurrence of HAT. Secondary endpoints were 1-year patient and graft survival determined by Kaplan-Meier analysis and complications. Multivariate analysis was used to identify independent risk factors for HAT and adverse events. A total of 79 pLTs were performed [30 (38.0%) living donations; 49 (62.0%) postmortem donations] divided into 23 (29.1%) segment 2/3, 32 (40.5%) left lobe, 4 (5.1%) extended right lobe, and 20 (25.3%) full-size grafts. One-year patient and graft survival were both 95.2% in the OM group versus 86.2% and 77.8% in the SL group (p = .276 and p = .077). HAT rate was 0% in the OM group versus 24.1% in the SL group (p = .013). One-year patient and graft survival were 64.3% and 35.7% in patient with HAT, compared to 93.9% and 92.8% in patients with no HAT (both p < .001). Multivariate analysis revealed HAR with SL (p = .022) and deceased donor liver transplantation (DDLT) (p = .014) as independent risk factors for HAT. The occurrence of HAT was independently associated with the need for retransplantation (p < .001) and biliary leakage (p = .045). In pLT, the use of an OM is significantly associated to reduce HAT rate, biliary complications, and graft loss and outweighs the disadvantages of delayed arterial perfusion and prolonged warm ischemia time (WIT).


Hypoxic cardiopulmonary-cerebral resuscitation fails to improve neurological outcome following cardiac arrest in dogs.
Hyperoxic cardiopulmonary resuscitation (CPR) is associated with an increase in neurologic dysfunction upon successful resuscitation with much of the damage attributable to an increase in reperfusion oxidant injury. We hypothesized that by contrast, hypoxic ventilation during resuscitation would improve neurologic outcome by reducing available substrate necessary for oxidant injury. Specifically, this study investigated the effects of 2 levels of hypoxic ventilation during resuscitation: F1O2 = 0.085, PaO2 = 26.6 +/- 3.4 mmHg, (HY8), and F1O2 = 0.12, PaO2 = 33.0 +/- 4.2 mmHg, (HY12), and normoxic resuscitation: F1O2 = 0.21, PaO2 = 60.6 +/- 17.0 mmHg, (N) on survival and neurological outcome following 9 min of normothermic cardiac arrest. Concentrations of malonaldehyde (MDA) and 4-hydroxynonenal (4-OH) in plasma and concentrations of glutathione (GSH) in erythrocyte lysates were measured to quantify possible radical damage. Physiological variables including arterial blood gases were followed for 24 h after resuscitation. Neurologic outcome was assessed using a standardized scoring system. Hypoxically (HY8) resuscitated dogs tended to have a greater neurologic deficit than normoxically resuscitated dogs and had reduced overall survival (16.9 +/- 8.9 h) compared to N dogs (24.0 +/- 0.0 h). Overall survival time correlated negatively (-0.693) and significantly (P = 0.0018) with plasma glucose concentration. Arterial plasma glucose concentrations were higher in the HY8 group compared to the N group immediately (HY8, 312 +/- 86 mg/dL; N, 196 +/- 82 mg/dL; P = 0.17) and 30 min (HY8, 331 +/- 109 mg/dL; N, 187 +/- 74 mg/dL; P = 0.077) following resuscitation. No statistically discernible differences in markers of oxidant injury were apparent among the 3 groups, but pooled data increased significantly with time for MDA and 4-OH. Pooled data for GSH showed a significant drop at 1 h following resuscitation and returned to normal by 6 h. Data from these markers suggested attendant oxidant injury in all groups. Thus, hypoxic ventilation at 2 depths of hypoxia during resuscitation failed to improve neurologic outcome beyond that achieved by ventilation with air, suggesting that normoxia rather than hyperoxia or hypoxia is the ideal target for arterial oxygenation during resuscitation.


The impact of enzyme characteristics on corn stover fiber degradation and acid production during ensiled storage.
Ensilage can be used to store lignocellulosic biomass before industrial bioprocessing. This study investigated the impacts of seven commercial enzyme mixtures derived from Aspergillus niger, Trichoderma reesei, and T. longibrachiatum. Treatments included three size grades of corn stover, two enzyme levels (1.67 and 5 IU/g dry matter based on hemicellulase), and various ratios of cellulase to hemicellulase (C:H). The highest C:H ratio tested, 2.38, derived from T. reesei, resulted in the most effective fermentation, with lactic acid as the dominant product. Enzymatic activity during storage may complement industrial pretreatment; creating synergies that could reduce total bioconversion costs.


Primary care management of child & adolescent depressive disorders.
This article outlines the importance of primary health care in addressing the public health challenge presented by pediatric depressive disorders. The current realities of depression management in primary care are discussed. The models emerging from intervention research and the barriers to their implementation in practice are then reviewed. Drawing on this background, recent new standards for primary care management of pediatric depressive disorders are discussed, along with resources that have been developed to support their achievement.


A pathogenic myxoma virus in vaccinated and non-vaccinated commercial rabbits.
A case of a myxoma virus strain in vaccinated and non-vaccinated rabbits is described, and genetic identification of that strain was performed in this study. In two commercial farms being 150km apart, myxomatosis has been occurred after the import of animals from a common supplier. The disease was manifested firstly in the existing non-immune population of does and fatteners, and later in all vaccinated animals, being 2-3 months immune at the time of first symptoms. Morbidity was almost 100% with nasal discharge, listlessness, fever, eyelid swelling, eye and nasal purulent discharge, papules in the ears, facial oedema, and swelling of the anagenital region, with result always the death of the animals. Examination by PCR had shown the presence of a 492-bp specific product in all the symptomatic animals tested from both farms, having 100% nucleotide sequence identity with the homologous region of the myxoma virus Lausanne strain. The simultaneous occurrence of myxomatosis in the vaccinated and non-vaccinated rabbits of both farms, suggests that the supplier was possibly the source of a viral isolate with increased virulence.


Fabrication of tumor-targeting composites based on the triple helical β-glucan through conjugation of aptamer.
Herein the nucleic acid aptamers were attached to the polydeoxyadenylic acid (poly(dA)) tail for improving the tumor-targetability and cellular internalization of s-LNT/poly(dA) composite composed of two single chains of triple helical β-glucan lentinan (s-LNT) and one poly(dA) chain. The in vitro results demonstrate that the cellular uptake of s-LNT/poly(dA) composites in MCF-7 cancer cells was enhanced effectively after attaching the aptamer. The as-prepared fluorescin isothiocyanate (FITC)-labelled LNT (LNT-FITC) through grafting was used for tracing the enhanced tumor-targetability of the composites. As a result, the cellular internalization of the LNT-FITC into MCF-7 and 4T1 cancer cells was further increased by the aptamer conjugated to poly(dA). Meanwhile, the in vivo experiments further demonstrate more s-LNT/poly(dA)-aptamer composites were effectively accumulated at the tumor site compared with s-LNT alone. This work provides a novel strategy for fabricating triplex β-glucan as delivery vectors with active tumor-targetability.


Theories of carcinogenesis and their impact on surgical treatment of colorectal cancer. A historical review.
From the time of Hippocrates until the late 19th century, physicians and surgeons were convinced that surgical attempts at treating colorectal cancers were doomed to failure. This opinion stemmed from prevailing views on carcinogenesis. The three dominant theories, the humoral, mineral, and lymph theories, held that all cancers developed in tissue that had a diseased disposition. Thus, excision of the gross tumor mass alone seemed unlikely to cure the patient. Consequently, surgical treatment of all cancers, and in particular colorectal cancer, was vehemently condemned. The 19th century represented a transition period. Advances in surgical technique made excision of rectal cancers feasible. Unfortunately, classical views that resection of cancer was futile delayed the development of surgical treatment for colorectal cancer. Indeed, it was not until the late 19th century that a few individuals ignored these tenets of classical medicine and attempted local resections of rectal cancers. By the second quarter of the 20th century, a radical change occurred in the prevailing theories of carcinogenesis. Wide acceptance of the unicellular origin of cancer and the mucosal origin of colorectal cancers washed away admonitions against surgical treatment of colorectal cancers. It became axiomatic that all cancers, including colorectal cancers, could be cured surgically if treated while still local diseases.


Beyond canonical models: why a broader understanding of Diptera-microbiota interactions is essential for vector-borne disease control.
Vector-borne diseases constitute a major global public health threat. The most significant arthropod disease vectors are predominantly comprised of members of the insect order Diptera (true flies), which have long been the focus of research into host-pathogen dynamics. Recent studies have revealed the underappreciated diversity and function of dipteran-associated gut microbial communities, with important implications for dipteran physiology, ecology, and pathogen transmission. However, the effective parameterization of these aspects into epidemiological models will require a comprehensive study of microbe-dipteran interactions across vectors and related species. Here, we synthesize recent research into microbial communities associated with major families of dipteran vectors and highlight the importance of development and expansion of experimentally tractable models across Diptera towards understanding the functional roles of the gut microbiota in modulating disease transmission. We then posit why further study of these and other dipteran insects is not only essential to a comprehensive understanding of how to integrate vector-microbiota interactions into existing epidemiological frameworks, but our understanding of the ecology and evolution of animal-microbe symbiosis more broadly.


Biological removal of phosphate at low concentrations using recombinant Escherichia coli expressing phosphate-binding protein in periplasmic space.
During wastewater treatment, phosphate removal is an important and challenging process; thus, diverse technologies, including those derived from biological means, have been devised for efficient phosphate removal. Although conventional biological methods are effective in decreasing wastewater phosphate levels to ~1 mg/L, long periods of microbial adaptation are required for effective phosphate removal, and the removal efficiency of these methods is relatively poor at lower phosphate concentrations. In the present work, we constructed a recombinant Escherichia coli with periplasmic-expressed phosphate-binding protein (PBP) and investigated its biological removal ability for low phosphate levels. We found that the PBP-expressing recombinant E. coli cells showed efficient (> 94 %) removal of phosphate at low concentrations (0.2-1.0 mg/L) in a treated cell mass-dependent manner. Collectively, we propose that our PBP-expressing recombinant whole-cell system could be successfully used during wastewater treatment for the biological removal of low concentrations of phosphate.


Corrigendum: Application of optical tweezers in cardiovascular research: More than just a measuring tool.
[This corrects the article DOI: 10.3389/fbioe.2022.947918.].


Early results of expanding the anatomical indications for using a Gore Iliac branch endoprosthesis to treat aortoiliac and iliac aneurysms.
To assess the safety and anatomical suitability of using a Gore Iliac Branch Endoprosthesis (IBE) in aortoiliac and iliac aneurysm repair. Between 2017 and 2020, 20 patients underwent endovascular aneurysm repair (EVAR) with a Gore IBE device (bilateral IBE, n = 1) after expanding the instructions for use (IFU) criteria. We evaluated the early clinical outcomes and suitability of the IFU criteria, retrospectively. Six patients (30%) met all the IFU criteria. Anatomical suitability according to the IFU criteria for the collective total of 21 IBE limbs was confirmed for 10 (47.6%) proximal common iliac arteries, 21 (100%) external iliac arteries, 18 (85.7%) internal iliac arteries, and in the length from the lowest renal artery to the iliac bifurcation in 15 (71.8%) patients. Assisted primary technical success was achieved in all patients with various bail-out techniques. One patient (5%) required a bare-stent insertion 7 days after EVAR for severe stenosis in the ipsilateral limb caused by a small terminal aorta. There was no case of occlusion of an iliac branch component device. Gore IBEs were implanted safely and effectively with various bail-out techniques to repair aortoiliac and iliac aneurysms in our Japanese patients with a low rate of inclusion IFU criteria.


[Unreliability of pulse oximetry in hypothermic children after cardiovascular surgery with deep hypothermic circulation].
The aim of the study was to evaluate the accuracy of oxygen saturation with the pulse oximeter (SpO2) in children undergoing cardiovascular surgery with deep hypothermic circulation. The SpO2 values measured at the hand and at the foot were compared in 50 patients with the arterial oxygen saturation (SaO2) measured with the oximeter. A variance value between the two measurements less than 3% was considered as a reliable measurement, and the precision of the measurement was defined as the standard deviation of the variability. For skin temperature between 35 and 28 C grade, and for core temperature more than 34 grade, the SpO2 is a reliable measurement of the SaO2; for skin temperature less than 28 grade and core temperature less than 34 grade the SpO2 do not correspond the SaO2. The variability and the precision of the measurements are not affected the position of the probe (hand or foot).


A randomized controlled trial to determine the effect of spinal stabilization exercise intervention based on pain level and standing balance differences in patients with low back pain.
A number of studies have evaluated exercise interventions compared with other treatment strategies for subjects with recurrent low back pain (LBP); however, subject pain level and balance were not carefully considered. The purpose of this study was to investigate the effectiveness of spinal stabilization exercises (SSE) for managing pain and increasing balance strategy changes following unexpected perturbations in patients diagnosed with recurrent LBP. Twenty-one age- and gender-matched patients participated in a supervised SSE or control exercise program 5 times a week over a 4-week period. The Million Visual Analogue Scale (MVAS) and Oswestry Disability Index (ODI) were used to measure each patient's level of pain and disability. Balance measurements were derived from recordings of the anterior-posterior (A/P) and medio-lateral (M/L) center of pressure (COP) displacements during 3 consecutive, unexpected random perturbations. The level of reported pain and disability significantly decreased following treatment for both groups. Although the M/L sway was not significantly different in either group (p=0.86), there was a significant difference between group and measurement time during A/P sway (p=0.04). The A/P displacement of the SSE group significantly decreased compared with the control group. The decreased A/P displacement can be linked to the SSE intervention, which helps prevent further injury by limiting an individual's response rate to external perturbations. Clinicians might consider SSE for LBP patients as a possible rehabilitation strategy to reduce A/P displacement.


Should all unrelated donors for transplantation be matched?
Previous analyses have suggested that hematopoietic cell transplantation (HCT) from an unrelated donor results in better survival if the patient is younger and, possibly also if the donor is younger. Additionally, survival is improved if HCT is performed during early disease stage and if the recipient and possibly the donor are cytomegalovirus (CMV) seronegative. Equivocal data have been published comparing bone marrow vs. (granulocyte-colony stimulating factor) G-CSF-stimulated peripheral blood stem cells for transplantation. A randomized trial is underway by the Blood and Marrow Transplant Clinical Trials Network that is testing the prospective comparison of bone marrow vs. primed peripheral blood grafts from unrelated donors and patients with hematologic malignancies. Of most significance, however, is that the best donor is HLA-compatible, healthy, promptly available, and willing to give the requested product for HCT.


Identification and molecular characterization of the interleukin-10 receptor 1 of the zebrafish (Danio rerio) and the goldfish (Carassius auratus L.).
This is the first report of the identification and molecular characterization of an interleukin-10 receptor 1 in bony fish. By gene synteny analysis, we identified the zebrafish interleukin-10 receptor 1 (IL10R1) and using this IL10R1 sequence, we cloned the goldfish IL10R1 cDNA transcript. The identified fish IL10R1 protein sequences had a putative JAK1 binding site, only one of the two STAT3 binding sites, that are present in all other vertebrates IL10R1 proteins as well as C-terminal serine rich areas, believed to be responsible for the anti-inflammatory properties of IL10R1. Phylogenetically, the fish IL10R1 proteins grouped independently of the amphibian, avian and mammalian IL10R1s. Quantitative gene expression analysis of the IL10R1 of zebrafish and goldfish revealed highest mRNA levels in the spleen tissues. High mRNA levels were also observed in the zebrafish muscle in contrast to low mRNA levels in the muscle of the goldfish. Moderate IL10R1 mRNA levels were seen in most other tissues examined and lowest gene expression was in the liver of both fish species. Goldfish monocytes stimulated with a recombinant goldfish interleukin-10 (rgIL-10) or with heat killed fish pathogens, Aeromonas salmonicida or Trypanosoma carassii, exhibited significantly reduced mRNA levels of the IL10R1. Furthermore, we produced a recombinant form of the goldfish IL10R1 (rgIL10R1) and using in vitro binding studies, demonstrated that the dimerized rgIL-10 specifically interacted with rgIL10R1. Our results suggest that interleukin-10 system has been highly conserved throughout evolution.


Working on the dirty side-the ipsilateral subclavian access for temporary pacing after lead extraction.
Temporary pacing is necessary in pacemaker-dependent patients after transvenous lead extraction (TLE) for cardiac implantable electronic device infection. In case of unavailability of other accesses, we propose to use the ipsilateral subclavian access (ISA) combined with a standard permanent active fixation lead for the temporary pacemaker and present preliminary data. We consecutively enrolled patients undergoing TLE who received a temporary pacemaker using the ISA between August 2016 and April 2020 at our centre. During the observation period, 36 patients undergoing TLE for pocket infection (72.2%), endocarditis (25.0%) or other causes received a temporary pacemaker over the ISA. Their mean age was 77.0 ± 10.7 years, and 13.9% were female. Complete TLE was achieved in 94.4%. There were no major periprocedural complications. Intra-hospital mortality was 11.1%. Pocket revision was performed in 19.4%. During long-term follow-up (23 ± 13 months), 8.3% had a relapse of local pocket infection and 2.8% needed rehospitalization for reintervention. Temporary pacing using a standard permanent active fixation lead using the ISA is a convenient alternative to conventional venous accesses. However, risks of implanting a lead into a previously infected area have to be taken into account.


Biomarkers of vascular calcification in serum.
Over the last decades, the association between vascular calcification (VC) and all-cause/cardiovascular mortality, especially in patients with high atherogenic status, such as those with diabetes and/or chronic kidney disease, has been repeatedly highlighted. For over a century, VC has been noted as a passive, degenerative, aging process without any treatment options. However, during the past decades, studies confirmed that mineralization of the arteries is an active, complex process, similar to bone genesis and formation. The main purpose of this review is to provide an update of the existing biomarkers of VC in serum and develop the various pathogenetic mechanisms underlying the calcification process, including the pivotal roles of matrix Gla protein, osteoprotegerin, bone morphogenetic proteins, fetuin-a, fibroblast growth-factor-23, osteocalcin, osteopontin, osteonectin, sclerostin, pyrophosphate, Smads, fibrillin-1 and carbonic anhydrase II.


Protection of Marine Mammals.
Within the European Defense Agency (EDA), the Protection of Marine Mammals (PoMM) project, a comprehensive common marine mammal database essential for risk mitigation tools, was established. The database, built on an extensive dataset collection with the focus on areas of operational interest for European navies, consists of annual and seasonal distribution and density maps, random and systematic sightings, an encyclopedia providing knowledge on the characteristics of 126 marine mammal species, data on marine mammal protection areas, and audio information including numerous examples of various vocalizations. Special investigations on marine mammal acoustics were carried out to improve the detection and classification capabilities.


Evaluation of the Therapeutic Efficacy of Perceptual Learning Using the Bartimeus Training Program in the Clinic as an Additional Therapy to Glasses or Patching.
The Bartimeus training program (BT) is a perceptual learning task that consists of a drawing assignment performed in 12 sessions of 30 minutes duration. Here we aim to retrospectively assess visual improvement in children who underwent BT and in whom visual acuity had not improved by glasses or patching in relation to patient age and diagnosis. Retrospective chart analysis of patients who had undergone BT at our institution after consent. Visual acuity prior to and following BT was analyzed, as were diagnosis and age at BT. Thirteen patients (20 eyes) with a mean age of 7 years (range: 3.9 - 10.75) were included. Overall acuity (LogMar) improved in 75%. Six patients aged 6.5 - 7.5 years had a mean improvement of 1.4 lines. The greatest improvement (5 lines crowding acuity) was seen in a 9-year-old with recent onset amblyopia due to corneal scarring. Two patients aged 6 years with deprivation amblyopia following surgery of a congenital cataract and one 5-year-old with retinitis pigmentosa did not benefit from BT. BT is a cheap noninvasive method that may help improve visual outcomes. Further studies with larger numbers are needed to assess optimal indication, timing, and length of treatment.


Research progress in mRNA drug modification and delivery systems.
Messenger RNA (mRNA) has shown tremendous potential in disease prevention and therapy. The clinical application requires mRNA with enhanced stability and high translation efficiency, ensuring it not to be degraded by nucleases and targeting to specific tissues and cells. mRNA immunogenicity can be reduced by nucleotide modification, and translation efficiency can be enhanced by codon optimization. The 5´ capping structure and 3´ poly A increase mRNA stability, and the addition of 5' and 3' non-translational regions regulate mRNA translation initiation and protein production. Nanoparticle delivery system protects mRNA from degradation by ubiquitous nucleases, enhances mRNA concentration in circulation and assists it cytoplasmic entrance for the purpose of treatment and prevention. Here, we review the recent advances of mRNA technology, discuss the methods and principles to enhance mRNA stability and translation efficiency; summarize the requirements involved in designing mRNA delivery systems with the potential for industrial translation and biomedical application. Furthermore, we provide insights into future directions of mRNA therapeutics to meet the needs for personalized precision medicine.


Effect of prostaglandin F2alpha at the time of AI on progesterone levels and pregnancy rate in synchronized Italian Mediterranean buffaloes.
The aims of this study were to evaluate the effects of an intravenous or intramuscular PGF2alpha analogue administration on the day of estrus on progesterone concentration and pregnancy rate in buffaloes undergoing artificial insemination (AI). To this end, two experiments were carried out. The first study was performed on 72 Mediterranean buffaloes synchronized by the Ovsynch-TAI Program. On the day of estrus only animals considered in heat were divided into four groups: Groups IVC and IMC received, respectively, an intravenous or intramuscular injection of cloprostenol (0.524 mg), whereas control Groups IVS and IMS received the same injections of saline. Milk samples were collected daily from each animal to assess progesterone concentration in the whey by RIA method. In addition on alternate days, buffaloes underwent transrectal ultrasound analysis. The second study was carried out on 385 buffaloes synchronized by the Ovsynch-TAI Program. On the day of AI, animals were divided in four groups, as described in experiment 1. Pregnancy rate was evaluated either on day 26 or day 45 and embryonic mortality rate was recorded. Statistical analysis was performed by ANOVA and chi2 test. A higher (P<0.05) progesterone concentration was recorded on day 11 (Day 0=estrus day) in Groups IVC and IMC compared to Groups IVS and IMS (351.6+/-129.7 and 355.8+/-112.2 pg ml(-1) vs. 239.8+/-81.1 and 243.6+/-90.5 pg ml(-1), respectively). Furthermore, a larger CL was recorded on the same day in treated vs. control groups (1.25+/-0.15 and 1.27+/-0.17 cm, respectively, in Groups IVC and IMC vs. 1.08+/-0.14 and 1.05+/-0.13 cm in IVS and IMS). In the second study, a higher pregnancy rate was observed in treated (IVC+IMC) vs. control (IVS+IMS) groups (46.7% vs. 30.7%; P<0.01), while no differences were recorded between treated groups. From these data, it can be concluded that either intravenous or intramuscular administration of PGF2alpha at the time of AI can enhance progesterone levels and pregnancy rate in buffaloes.


Effects of lipid peroxides on prostacyclin and thromboxane generation in hypercholesterolemic rabbits.
Rabbits fed an atherogenic diet for 60 days resulted in high levels of plasma lipid peroxides as well as extreme hypercholesterolemia. Both levels stayed high until 35 days after the atherogenic diet stopped. At the same time, plasma PGI2 level was remarkably decreased while TXA2 and platelet aggregability were increased. Atherosclerotic aortas contain high levels of lipid peroxides associated with decreased PGI2 and increased TXA2 generation. Atherosclerotic plaques had the highest level of lipid peroxides and TXA2 while PGI2 production was the least, as compared with nonplaque tissue of the same artery and the normal arteries. The condition of normal arteries was just the reverse. There was a negative correlation between lipid peroxides and prostacyclin production, and a positive correlation between lipid peroxides and TXA2, in both plasma and aorta of rabbits. These results suggest that there is a close correlation between atherosclerosis, elevated lipid peroxides, and disturbances in PGI2/TXA2 balances.


PAR1 inhibition suppresses the self-renewal and growth of A2B5-defined glioma progenitor cells and their derived gliomas in vivo.
Glioblastoma (GBM) remains the most common and lethal intracranial tumor. In a comparison of gene expression by A2B5-defined tumor-initiating progenitor cells (TPCs) to glial progenitor cells derived from normal adult human brain, we found that the F2R gene encoding PAR1 was differentially overexpressed by A2B5-sorted TPCs isolated from gliomas at all stages of malignant development. In this study, we asked if PAR1 is causally associated with glioma progression. Lentiviral knockdown of PAR1 inhibited the expansion and self-renewal of human GBM-derived A2B5(+) TPCs in vitro, while pharmacological inhibition of PAR 1 similarly slowed both the growth and migration of A2B5(+) TPCs in culture. In addition, PAR1 silencing potently suppressed tumor expansion in vivo, and significantly prolonged the survival of mice following intracranial transplantation of human TPCs. These data strongly suggest the importance of PAR1 to the self-renewal and tumorigenicity of A2B5-defined glioma TPCs; as such, the abrogation of PAR1-dependent signaling pathways may prove a promising strategy for gliomas.


Host location by ichneumonid parasitoids is associated with nest dimensions of the host bee species.
Parasitoid fitness depends on the ability of females to locate a host. In some species of Ichneumonoidea, female parasitoids detect potential hosts through vibratory cues emanating from them or through vibrational sounding produced by antennal tapping on the substrate. In this study, we (1) describe host location behaviors in Grotea gayi Spinola (Hymenoptera: Ichneumonidae) and Labena sp. on nests of Manuelia postica Spinola (Hymenoptera: Apidae), (2) compare nest dimensions between parasitized and unparasitized nests, (3) correlate the length of M. postica nests with the number of immature individuals developing, and (4) establish the relative proportion of parasitized nests along the breeding period of M. postica. Based on our results, we propose that these parasitoids use vibrational sounding as a host location mechanism and that they are able to assess host nest dimensions and choose those which may provide them with a higher fitness. Finally, we discuss an ancestral host-parasitoid relationship between Manuelia and ichneumonid species.


Alteration by prolactin of surface charge and membrane fluidity of rat 13762 mammary ascites tumor cells.
Intraperitoneal injection of ovine prolactin (100 micrograms/d) in Fischer 344 rats bearing transplantable 13762 mammary ascites tumor (MAT) cells modifies the surface charge density and membrane fluidity of the tumor cells. In each of five experiments the mean electrophoretic mobility (epm) of MAT cells taken from prolactin-treated rats was significantly lower than that of cells from nonhormone-treated controls. Prolactin concentrations were increased in vivo by (a) direct intraperitoneal injection of ovine prolactin; (b) subcutaneous implantation of diethylstilbestrol-containing silastic capsules to produce pituitary prolactin secreting tumors; or (c) a single subcutaneous injection of polyestradiol phosphate, a long-acting estrogen. In an effort to establish that the prolactin effect was a direct one, two in vivo protocols were used: (a) MAT cells were coincubated with anterior pituitary halves obtained from nontumor-bearing littermates; or (b) rat or ovine prolactin was added to serum-free culture media containing MAT cells. In both protocols, the epm of the prolactin-treated cells was significantly lower. The isoelectric focusing pH of whole cells was increased by prolactin treatment from 4.93 to 5.12, consistent with a reduction in the number of surface carboxyl groups. The fluidity of membranes of treated cells was drastically increased, as measured by spin-label probe rotation rates. These combined results imply that the hormone exerts its effect by stimulating events in the cell that lead to a reduction of the average density of carboxylic acid residues on the tumor cell surface.


Melt-spun microbial poly(3-hydroxybutyrate-co-3-hydroxyvalerate) fibers with enhanced toughness: Synergistic effect of heterogeneous nucleation, long-chain branching and drawing process.
A series of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) composite fibers were prepared via melt reactive processing where tungsten disulphide (WS2) and dicumyl peroxide (DCP) was used as heterogeneous nucleating agent and initiator, respectively. Compared with PHBV or PHBV/WS2 fiber, the resultant LCB-PHBV/WS2 fibers showed improving mechanical performance, a higher nucleation temperature and a rapid crystallization rate due to the synergistic effect between heterogeneous nucleation, long chain branched structure and draw-induced crystallization. Taking the results with a cooling rate of 20 °C/min as an example, the peak crystallization temperature of LCB-PHBV/1.0 wt%WS2 increased from 100.47 °C to 107.40 °C and the crystallization enthalpy increased from 87.95 J/g to 93.03 J/g. Moreover, two different crystalline forms, α-crystal and β-crystal, were found in the prepared PHBV fibers, and the β-crystal part increased with the inducement of the stretching ratio which enhanced the fiber strength, given the tensile strength and elongation at break of the LCB-PHBV/WS2 fibers up to 189.8 MPa and 46.5%, respectively.


Ion specificity and ionic strength dependence of the osmoregulatory ABC transporter OpuA.
The ATPase subunit of the osmoregulatory ATP-binding cassette transporter OpuA from Lactococcus lactis has a C-terminal extension, the tandem cystathionine beta-synthase (CBS) domain, which constitutes the sensor that allows the transporter to sense and respond to osmotic stress (Biemans-Oldehinkel, E., Mahmood, N. A. B. N., and Poolman, B. (2006) Proc. Natl. Acad. Sci. U. S. A. 103, 10624-10629). C-terminal of the tandem CBS domain is an 18-residue anionic tail (DIPDEDEVEEIEKEEENK). To investigate the ion specificity of the full transporter, we probed the activity of inside-out reconstituted wild-type OpuA and the anionic tail deletion mutant OpuADelta12; these molecules have the tandem CBS domains facing the external medium. At a mole fraction of 40% of anionic lipids in the membrane, the threshold ionic strength for activation of OpuA was approximately 0.15, irrespective of the electrolyte composition of the medium. At equivalent concentrations, bivalent cations (Mg(2+) and Ba(2+)) were more effective in activating OpuA than NH(4)(+), K(+), Na(+), or Li(+), consistent with an ionic strength-based sensing mechanism. Surprisingly, Rb(+) and Cs(+) were potent inhibitors of wild-type OpuA, and 0.1 mM RbCl was sufficient to completely inhibit the transporter even in the presence of 0.2 M KCl. Rb(+) and Cs(+) were no longer inhibitory in OpuADelta12, indicating that the anionic C-terminal tail participates in the formation of a binding site for large alkali metal ions. Compared with OpuADelta12, wild-type OpuA required substantially less potassium ions (the dominant ion under physiological conditions) for activation. Our data lend new support for the contention that the CBS module in OpuA constitutes the ionic strength sensor whose activity is modulated by the C-terminal anionic tail.


Effect and mechanism of Tangzhiqing in improving cardiac function in mice with hyperlipidaemia complicated with myocardial ischaemia.
Tangzhiqing formula (TZQ) is a traditional Chinese medicine prescribed to treat lipid metabolism disorders, atherosclerosis, diabetes and diabetic cardiomyopathy. However, some challenges and hurdles remain. TZQ showed promising results in treating diabetes and hyperlipidaemia. However, its effect on and mechanism of action in hyperlipidaemia complicated with myocardial ischaemia (HL-MI) remain unknown. In this study, a network pharmacology-based strategy integrating target prediction was adopted to predict the targets of TZQ relevant to the treatment of HL-MI and to further explore the involved pharmacological mechanisms. A total of 104 potential therapeutic targets were obtained, including MMP9, Bcl-2, and Bax, which may be related to the apoptosis and PI3K/AKT signalling pathways. Then, we confirmed these potential targets and pathways with animal experimentation. TZQ reduced lipid levels, increased the expression levels of Bcl-2, decreased Bax, caspase-3 and caspase-9 expression levels, and activated the PI3K/AKT signalling pathway. In conclusion, this study provides new insights into the protective mechanisms of TZQ against HL-MI through network pharmacology and pharmacological approaches.


Improved sperm concentration, motility, and fertilization rates following Ficoll treatment of sperm in a human in vitro fertilization program.
Duplicate sperm samples from the spouses of 54 patients admitted to an in vitro fertilization program were prepared by the swim-up and a simplified procedure using Ficoll. Cellsoft (CRYO Resources, New York, NY) sperm curvilinear velocities (microns/sec) and mean amplitude of lateral head displacement values (microns) equivalent to grade 1 and 2 visual sperm motility were significantly higher for Ficoll as compared with swim-up samples (P less than 0.01). Fertilization rates were significantly higher in the Ficoll as compared with the swim-up group for poor semen samples (grade less than 2: 58% versus 24%; P less than 0.01) and normal semen samples (grade greater than or equal to 2: 85% versus 78%; P less than 0.05). Ficoll sperm separation appears to be an excellent method of yielding increased fertilization rates in in vitro fertilization programs.


Reduced parietal activation in participants with mild cognitive impairments during visual-spatial processing measured with functional near-infrared spectroscopy.
Functional Near Infrared Spectroscopy (fNIRS) may be a suitable, simple, and cost-effective brain imaging technique for detecting divergent neuronal patterns at an early stage of neurodegeneration. In course of Mild Cognitive Impairment (MCI) or Alzheimer's disease (AD), a deficit in visual-spatial processing, located in the parietal cortex, is a reliable risk factor. Earlier, we established the application of the clock-hand-angle-discrimination task (ADT) during fNIRS to identify neuronal correlates of the visual-spatial processing in a healthy elderly sample. In this analysis, we aimed to measure and find out differences in the hemodynamic response in MCI participants compared to matched healthy controls. As expected, MCI participants showed more errors over all conditions of pointer length and a higher reaction time in the long and middle pointer length condition. Moreover, results revealed a significant reduction of cortical activation in MCI patients. There was a generally increased activity in both the right as compared to the left hemisphere and the superior parietal brain region as compared to the inferior parietal brain region in both groups. In summary, fNIRS can be implemented in the measurement of visual-spatial processing in MCI patients and healthy elderly based on ADT. MCI participants had difficulties to cope with the ADT. Since neuronal hypoactivity occurs with concomitant behavioral deficits, an additional analysis was performed on a subgroup of MCI patients who performed as well as the control group in behavior. This subgroup analysis also showed a hypoactivation of the parietal cortex, without evidence of a compensatory activation. Therefore, we assume that MCI patients are characterized by a deficit in the parietal cortex. Overall, these findings confirm our hypothesis that hemodynamic deficits in visual-spatial processing, localized in the parietal cortex, are reliable and early diagnostic markers for cognitive decline in risk groups for the development of AD.


An electron-microscopic study of avirulent and virulent Semliki forest virus in the brains of different ages of mice.
Ultrastructural studies of brains infected with avirulent and virulent strains of Semliki forest virus (SFV) were performed in 2-7, 14, 19, and 21-28 day old mice. Mature virus particles, dense clumps of fine granules, spherules and advanced stages of viral development i.e. cytopathic vacuoles, Type II (CPV II) are seen in the brains at all ages with the virulent strain which is pathogenic to all age group of mice. In the avirulent strain infection which is pathogenic to mice below 15 days old, no mature virus particles or advanced stages of viral development are seen in 19 day old and adult mice in spite of high virus titres. However, dense clumps of fine granules and spherules are seen which seem to have the capacity to develop into lethal highly infectious mature virus when reinoculated into 2-7 day old mice. It is suggested that the dense clumps of fine granules and spherules are very early viral forms, which are seen in all age groups of mice. The cut-off mechanism in pathogenicity of the avirulent strain occurs in mice around 14 days old. Though some mature virus particles and advanced viral developmental stages are seen in these mice they were much less frequent compared to baby mice. Lymphoblastic type cells are numerous in this age group. These cells are seen more frequently in all ages of mice infected with the avirulent strain as compared to the virulent strain. These mononuclear cells can have an immunological role and may play a part in limiting the process of viral maturation and hence preventing death. Polymorphs and macrophages are rare with the avirulent infection but are the predominant infiltrating cells in the virulent strain infections. With the avirulent infections the astrocytes show hypertrophy and intranuclear inclusions are seen in them.


Understanding delusions.
Delusions traditionally have been considered as fixed, false beliefs, born of morbidity. Whereas this definition serves to orient the clinician to the phenomena at hand, each element breaks down under scrutiny. It has been shown that delusions are not necessarily false, although in some sense they are discordant with reality. When delusions coincide with actual events their judgements can be shown to be independent of this evidential basis; when they refer to disorders of experience, such as first rank symptoms, the experience usually contains a distorted meaning. The supposition that delusions are a variety of belief has itself been questioned. On the one hand, they do not always refer in a meaningful way to anything, or when they do they fail to function as evaluative judgments; instead, delusions are experienced subjectively in ways that are characteristic of knowing rather than believing. On the other hand, delusions are not ascertained clinically by surveying the patient's belief system; rather their failure to achieve the status of objective knowledge leads to the post hoc relegation of delusions to the epistemologic waste basket of beliefs. To treat delusions as necessarily the product of morbidity is essentially tautologous insofar as delusions are, by definition, pathologic; that is, as defective judgments delusions are not simply erroneous, they are disordered. Finally, the fixity of delusions is an empirical matter and varies widely. Underlying this perceived intractability, however, are the subjective certainty and incorrigibility that Jaspers identified and which Spitzer has recast in the form of "epistemological asymmetry" misapplied to external reality. Although delusions typically have been recognized and categorized according to their manifest content, these formal considerations are crucial to understanding the nature of delusions.


What do people really think about grief counseling? Examining community attitudes.
To understand why bereaved people who are highly distressed significantly underutilize grief counseling and therapy, we surveyed adults (N = 156) to investigate community attitudes toward grief counseling. Overall attitude was positive and women reported a significantly more positive attitude than men; there were no associations between attitude and age, country of birth, previous counseling, or bereavement experience. After controlling for gender, beliefs and affects explained attitudes toward grief counseling; behavioral responses did not. Targeting beliefs about grief counseling may promote positive attitudes so that people most likely to benefit from intervention will be more likely to seek it.


Understanding catalytic synergy in dinuclear polymerization catalysts for sustainable polymers.
Understanding the chemistry underpinning intermetallic synergy and the discovery of generally applicable structure-performances relationships are major challenges in catalysis. Additionally, high-performance catalysts using earth-abundant, non-toxic and inexpensive elements must be prioritised. Here, a series of heterodinuclear catalysts of the form Co(III)M(I/II), where M(I/II) = Na(I), K(I), Ca(II), Sr(II), Ba(II) are evaluated for three different polymerizations, by assessment of rate constants, turn over frequencies, polymer selectivity and control. This allows for comparisons of performances both within and between catalysts containing Group I and II metals for CO2/propene oxide ring-opening copolymerization (ROCOP), propene oxide/phthalic anhydride ROCOP and lactide ring-opening polymerization (ROP). The data reveal new structure-performance correlations that apply across all the different polymerizations: catalysts featuring s-block metals of lower Lewis acidity show higher rates and selectivity. The epoxide/heterocumulene ROCOPs both show exponential activity increases (vs. Lewis acidity, measured by the pKa of [M(OH2)m]n+), whilst the lactide ROP activity and CO2/epoxide selectivity show linear increases. Such clear structure-activity/selectivity correlations are very unusual, yet are fully rationalised by the polymerization mechanisms and the chemistry of the catalytic intermediates. The general applicability across three different polymerizations is significant for future exploitation of catalytic synergy and provides a framework to improve other catalysts.


Myocardial infarction with normal coronary arteries complicated by rupture of the left ventricular free wall.
We report the case of a 73-year-old woman successfully treated for a subacute rupture of the ventricular free wall which occurred on the fourth day after a postero-lateral myocardial infarction. Angiography performed prior to surgical repair revealed the presence of normal coronary arteries. The pathogenetic mechanism of such a happening remains uncertain, but the role of abrupt recanalization must be considered.


Intersectional stigma subgroup differences in unhealthy drinking and disordered marijuana use among Black and Latino cisgender sexual minority young men.
We investigated associations of intersectional stigma subgroups with alcohol and marijuana use among Black and Latino sexual minority young men. Subgroups included Minimal Stigma (low to no stigma), Select Social Stigma (occasional stigma in social relationships), Multiform Heterosexism (internalized and interpersonal heterosexism from family/friends), Multiform Racism (racism across diverse contexts), Compound Stigma (frequent, ubiquitous racism and heterosexism). Cohort of Black and Latino cisgender sexual minority young men (n = 414; baseline ages 16-25) surveyed semiannually 2016-2019. Generalized estimating equations integrated with latent class analysis modeled linear and quadratic age effects and association of stigma subgroups with past 6-month alcohol use, marijuana use, unhealthy drinking, and marijuana use disorder symptoms. All past 6-month substance use peaked between ages 21-23 years old. Across all ages and relative to Minimal Stigma, odds of drinking were higher in every subgroup and highest in Compound Stigma (OR=2.72, 95% CI 1.17-6.35); unhealthy drinking was higher in every subgroup and highest in Multiform Heterosexism (β = 3.31, 95% CI 1.92-3.89); marijuana use disorder symptoms were higher in most subgroups and highest in Compound Stigma (β = 1.30, 95% CI 0.76-1.85). Marijuana use odds did not differ among groups. By examining intersectional stigma subgroups, we identified subgroups for whom substance use was elevated during a development period when use tends to be highest. Young men experiencing stigma patterns characterized primarily by heterosexism or heterosexism together with racism may be especially at risk for developing unhealthy drinking behaviors and marijuana use disorder symptoms.


The site and receptors responsible for the inhibition by sympathetic nerves of intestinal smooth muscle and its parasympathetic motor nerves.
1. The effects of three inhibitory stimuli, sympathetic nerve stimulation, noradrenaline (NA) and isoprenaline have been examined on three forms of motor activity in the rabbit colon, the response to pelvic (parasympathetic) nerve stimulation, acetylcholine (ACh) and spontaneous tone.2. The response to pelvic nerve stimulation is most effectively inhibited by sympathetic nerve stimulation, much less effectively by NA and hardly at all by isoprenaline. The sympathetic nerves can inhibit the pelvic response at frequencies of stimulation which do not affect spontaneous tone. The inhibitory effect of sympathetic stimulation, and of NA, on the pelvic response is reduced by phentolamine 5 x 10(-6)M and unaffected by propranolol 5 x 10(-6)M suggesting the effect is mediated via alpha receptors.3. The response to ACh is inhibited by all three stimuli equally. The inhibitory effect of sympathetic nerve stimulation and of isoprenaline is reduced by propranolol 5 x 10(-6)M. The inhibitory effect of NA is also reduced by propranolol but to a lesser extent. Phentolamine 5 x 10(-6)M has a small effect in reducing the inhibitory effect of sympathetic nerve stimulation or of NA. This effect of phentolamine is lost if the participation of motor nerves in the response to ACh is excluded by either tetrodotoxin 10(-7) g/ml. or cold storage for 10-14 days. These results suggest that inhibition of the ACh response takes place mainly at the muscle by activation of beta receptors but that ACh may have a small indirect stimulant action through motor nerves and this is susceptible to inhibition through alpha receptors.4. All three stimuli are equally effective in lowering smooth muscle tone. This inhibitory effect of sympathetic nerve stimulation and of isoprenaline is reduced by propranolol 5 x 10(-6)M and unaffected by phentolamine 5 x 10(-6)M. The inhibitory effect of NA is reduced by propranolol but again is less sensitive to block than the other two inhibitory stimuli. Phentolamine is without effect on the inhibitory action of NA and the combination of phentolamine with propranolol is no more effective than propranolol alone. These results suggest that NA liberated by sympathetic nerves and isoprenaline inhibit myogenic tone in the smooth muscle by an action on beta receptors but the action of NA added to the bath cannot be fully explained in this way.


Targeted ubiquitination of sensory neuron calcium channels reduces the development of neuropathic pain.
Neuropathic pain caused by lesions to somatosensory neurons due to injury or disease is a widespread public health problem that is inadequately managed by small-molecule therapeutics due to incomplete pain relief and devastating side effects. Genetically encoded molecules capable of interrupting nociception have the potential to confer long-lasting analgesia with minimal off-target effects. Here, we utilize a targeted ubiquitination approach to achieve a unique posttranslational functional knockdown of high-voltage-activated calcium channels (HVACCs) that are obligatory for neurotransmission in dorsal root ganglion (DRG) neurons. CaV-aβlator comprises a nanobody targeted to CaV channel cytosolic auxiliary β subunits fused to the catalytic HECT domain of the Nedd4-2 E3 ubiquitin ligase. Subcutaneous injection of adeno-associated virus serotype 9 encoding CaV-aβlator in the hind paw of mice resulted in the expression of the protein in a subset of DRG neurons that displayed a concomitant ablation of CaV currents and also led to an increase in the frequency of spontaneous inhibitory postsynaptic currents in the dorsal horn of the spinal cord. Mice subjected to spare nerve injury displayed a characteristic long-lasting mechanical, thermal, and cold hyperalgesia underlain by a dramatic increase in coordinated phasic firing of DRG neurons as reported by in vivo Ca2+ spike recordings. CaV-aβlator significantly dampened the integrated Ca2+ spike activity and the hyperalgesia in response to nerve injury. The results advance the principle of targeting HVACCs as a gene therapy for neuropathic pain and demonstrate the therapeutic potential of posttranslational functional knockdown of ion channels achieved by exploiting the ubiquitin-proteasome system.


Nasal carriage of Staphylococcus aureus and cross-contamination in a surgical intensive care unit: efficacy of mupirocin ointment.
A six month prospective study was carried out in a surgical intensive care unit (SICU) of a university hospital to assess the incidence and routes of exogenous colonization by Staphylococcus aureus. A total of 157 patients were included in the study. One thousand one hundred and eleven specimens (nasal, surgical wound swabs, tracheal secretions obtained on admission and once a week thereafter, and all clinical specimens) were collected over a four month period from patients without nasal decontamination (A). They were compared with 729 specimens collected over a two month period from patients treated with nasal mupirocin ointment (B). All S. aureus strains were typed by restriction fragment length polymorphism (RFLP) pulsed-field gel electrophoresis after SmaI macrorestriction. The nasal colonization rates on admission were 25.5 and 32.7% in groups A and B, respectively. Thirty-one untreated patients (31.3%) and three patients (5.1%) treated with nasal ointment, acquired the nasal S. aureus in the SICU (P = 0.00027). Nasal carriers were more frequently colonized in the bronchopulmonary tract (Bp) and surgical wound (Sw) (62%) than patients who were not nasal carriers (14%) (P < 0.00001). The patterns were identical for nasal, Bp and Sw strains from the same patient. RFLP analysis characterized seven epidemic strains of methicillin-resistant S. aureus (MRSA) which colonized 60% of group A and 9% of group B patients (P < 0.00001). The bronchopulmonary tract infection rate was reduced in group B (P = 0.032). In conclusion, in an SICU, nasal carriage of S. aureus appeared to be the source of endogenous and cross-colonization. The use of nasal mupirocin ointment reduced the incidence of Bp and Sw colonization, as well as the MRSA infection rate.


Brachytherapy for T1-T2 floor-of-the-mouth cancers: the Gustave-Roussy Institute experience.
In a retrospective analysis, we evaluated the Gustave-Roussy Institute's experience of locoregional control, survival, and complications of low-dose rate brachytherapy for carcinoma of the floor of the mouth. Between 1970 and 1985, 160 patients with previously untreated carcinoma of the floor of the mouth received interstitial brachytherapy as definitive treatment. Of the 160 patients, 79 (49%) had T1 and 81 (51%) had T2 lesions, and 127 (79%) had N0 and 33 (21%) had N1; 84% of tumors arose from the anterior floor of the mouth. Brachytherapy was performed with 192Ir wires, according to the Paris system rules, followed by neck dissection (T2 or N1) or follow-up (T1N0). With a follow-up period of 9-19 years, the observed survival rates were 89% at 2 years and 76% at 5 years, and the local control rates were 93% in T1 and 88% in T2 tumors. A low rate of distant metastases was noticed (5%); 31% of patients developed a second primary cancer. Severe mucosal necrosis was observed in <10% of patients. Any grade of bone necrosis was seen in 18% of cases (only 2.5% had G3 necrosis). This complication occurred more frequently in patients with poor dental status and in those treated without dental protection during implantation (p <0.001). Radical brachytherapy offers excellent local control (89%) and an acceptable rate of complications (<10% severe necrosis) that may be significantly decreased with dental care and the use of protective devices. The high incidence of second malignancies remains a major concern in these patients.


The arginine-rich RNA-binding motif of HIV-1 Rev is intrinsically disordered and folds upon RRE binding.
Arginine-rich motifs (ARMs) capable of binding diverse RNA structures play critical roles in transcription, translation, RNA trafficking, and RNA packaging. The regulatory HIV-1 protein Rev is essential for viral replication and belongs to the ARM family of RNA-binding proteins. During the early stages of the HIV-1 life cycle, incompletely spliced and full-length viral mRNAs are very inefficiently recognized by the splicing machinery of the host cell and are subject to degradation in the cell nucleus. These transcripts harbor the Rev Response Element (RRE), which orchestrates the interaction with the Rev ARM and the successive Rev-dependent mRNA export pathway. Based on established criteria for predicting intrinsic disorder, such as hydropathy, combined with significant net charge, the very basic primary sequences of ARMs are expected to adopt coil-like structures. Thus, we initiated this study to investigate the conformational changes of the Rev ARM associated with RNA binding. We used multidimensional NMR and circular dichroism spectroscopy to monitor the observed structural transitions, and described the conformational landscapes using statistical ensemble and molecular-dynamics simulations. The combined spectroscopic and simulated results imply that the Rev ARM is intrinsically disordered not only as an isolated peptide but also when it is embedded into an oligomerization-deficient Rev mutant. RRE recognition triggers a crucial coil-to-helix transition employing an induced-fit mechanism.


Internal States Influence the Representation and Modulation of Food Intake by Subthalamic Neurons.
Deep brain stimulation of the subthalamic nucleus (STN) is an effective therapy for motor deficits in Parkinson's disease (PD), but commonly causes weight gain in late-phase PD patients probably by increasing feeding motivation. It is unclear how STN neurons represent and modulate feeding behavior in different internal states. In the present study, we found that feeding caused a robust activation of STN neurons in mice (GCaMP6 signal increased by 48.4% ± 7.2%, n = 9, P = 0.0003), and the extent varied with the size, valence, and palatability of food, but not with the repetition of feeding. Interestingly, energy deprivation increased the spontaneous firing rate (8.5 ± 1.5 Hz, n = 17, versus 4.7 ± 0.7 Hz, n = 18, P = 0.03) and the depolarization-induced spikes in STN neurons, as well as enhanced the STN responses to feeding. Optogenetic experiments revealed that stimulation and inhibition of STN neurons respectively reduced (by 11% ± 6%, n = 6, P = 0.02) and enhanced (by 36% ± 15%, n = 7, P = 0.03) food intake only in the dark phase. In conclusion, our results support the hypothesis that STN neurons are activated by feeding behavior, depending on energy homeostatic status and the palatability of food, and modulation of these neurons is sufficient to regulate food intake.


Oscillatory activity in the occipitotemporal area related to the visual perception of letters of a first/second language and pseudoletters.
The objective of this study was to reveal the oscillatory activity in the occipitotemporal area related to the visual perception of the letters of first (L1) and second languages (L2) and pseudoletters. We recorded neuromagnetic signals while Korean native speakers were exposed to a phonogram of Korean, acquired at school age as their L1 (Hangul), that of Japanese, learned in adulthood as a L2 (Kana) and pseudoletter (Pseudo), and quantified the event-related desynchronization (ERD) and synchronization (ERS). In all conditions, sustained ERDs in the alpha band were observed in both hemispheres. ERD for Pseudo was gradually attenuated after approximately 400-500 ms after stimulus onset, whereas both Hangul and Kana produced stronger and longer-lasting ERD. ERD for Kana showed a broader alpha band than Hangul. Furthermore, transient ERSs in the gamma band around 70 Hz were observed between 100 and 400 ms in the bilateral occipitotemporal areas. In the left hemisphere, gamma band oscillations showed similar enhancement in all conditions, suggesting that gamma band activity in the left occipitotemporal area might be enhanced not only by the bottom-up process as visual perception but also by the top-down process as attention to prelexical visual stimuli. In the right hemisphere, gamma band ERS was stronger for Hangul than Pseudo and no differences were shown between Kana and Pseudo. The differences of oscillatory activity in the alpha and gamma bands suggest that neuronal networks, including the occipitotemporal area, are related to the visual perception of letters differing between L1 and L2.


Robust interferon signature and suppressed tissue repair gene expression in synovial tissue from patients with postinfectious, Borrelia burgdorferi-induced Lyme arthritis.
In most patients with Lyme arthritis (LA), antibiotic therapy results in Borrelia burgdorferi pathogen elimination, tissue repair, and return to homeostasis. However, despite spirochetal killing, some patients develop proliferative synovitis, characterised by synovial hyperplasia, inflammation, vascular damage, and fibrosis that persists for months to several years after antibiotic treatment, called postinfectious LA. In this study, we characterised the transcriptomes of postinfectious LA patients' synovial tissue, the target tissue of the immune response. High-throughput RNA sequencing to a depth of ~30 million reads per sample was used to profile gene expression in synovial tissue from 14 patients with postinfectious LA, compared with eight patients with other types of chronic inflammatory arthritis and five with minimally inflammatory osteoarthritis (OA). Synovium from postinfectious LA and other inflammatory arthritides shared gene signatures associated with antigen presentation, innate immune responses, and cell-mediated immune activation, whereas these responses were diminished in OA synovium. Unique to postinfectious LA was a particularly robust interferon-gamma (IFNγ) signature. Moreover, this heightened IFNγ signature inversely correlated with expression of genes involved in repair of damaged tissue, including genes associated with stromal cell proliferation and differentiation, neovascularisation, and extracellular matrix synthesis, which were markedly suppressed in postinfectious LA. Transcriptional observations were confirmed by cytokine profiling, histologic analyses, and clinical correlations. We propose that in patients with postinfectious LA, overexpression of IFNγ in synovium prevents appropriate repair of tissue damaged by B. burgdorferi infection, blocking return to tissue homeostasis long after completion of antibiotic therapy and resolution of active infection.


Electrocatalytic Behavior of Hemoglobin Oxidation of Hydrazine Based on ZnO Nano-rods with Carbon Nanofiber Modified Electrode.
A novel biosensor was developed by immobilizing hemoglobin (Hb) on a glassy carbon electrode (GCE) modified with a composite of ZnO nano-rods and carbon nanofiber (CNF), a strong reducer, hydrazine, was firstly used to evaluate the electrochemical behavior of Hb on Hb/ZnO/CNF/GCE. UV-vis and circular dichroism (CD) spectra indicated the conformational structure of Hb interaction with ZnO/CNF was predominantly an α-helical structure. The modified electrodes were characterized by scanning electron microscopy (SEM), electron impedance spectroscopy (EIS), and cyclic voltammetry. Electrocatalytic mechanism of Hb to oxidation reaction of hydrazine was suggested. The bioelectrocatalytic activity, kinetic parameters of Michaelis-Menten constant (Km), stability and reproducibility were also investigated. A linear dependence of peak currents to the concentrations of hydrazine was observed in the range from 1.98 × 10(-5) to 1.71 × 10(-3) mol L(-1) with a correlation coefficient of 0.998, and a detection limit (S/N = 3) of 6.60 μmol L(-1) was estimated.


Organization and complete nucleotide sequence of the gene encoding mouse phenylethanolamine N-methyltransferase.
Phenylethanolamine N-methyltransferase (PNMT; EC 188.8.131.52) catalyzes the conversion of norepinephrine to epinephrine, the last step of catecholamine biosynthesis. We have previously reported molecular cloning of cDNA encoding human PNMT and chromosomal localization of its gene (Kaneda et al., J. Biol. Chem., 263 (1988) 7672-7677). In this report, we isolated the chromosomal gene encoding mouse PNMT by cross-hybridization with the human PNMT cDNA. Mouse PNMT gene spanned about 1.8 kb and consisted of 3 exons. Primer extension analysis showed two putative transcription initiation sites. Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of the mouse PNMT mRNA in brain (pons and medulla oblongata) and adrenal gland. Subsequently cDNA encoding mouse PNMT was amplified by RT-PCR and cloned into the plasmid vector. Mouse PMNT gene contained the protein-coding region of 885 bp (295 amino acids) with the predicted molecular weight of 32,627. The deduced amino acid sequence of mouse PNMT revealed the major difference in the N-terminal region, as compared to the human and bovine PNMT sequences. In the 5'-terminal region of the mouse PNMT gene, we found the existence of 23 bp direct repeat sequences, which was not observed in the corresponding regions of the human and bovine PNMT genes. The presence or absence of the direct repeats caused the major difference in the PNMT sequences among species. The typical TATA, GC, and CACCC boxes as well as several sequences homologous to glucocorticoids response elements (GRE) were located in the 5'-flanking region of the mouse PNMT gene.


Blood loss in transurethral prostatectomy: epidural versus general anaesthesia.
The peroperative and postoperative blood loss was determined in 36 patients undergoing transurethral prostatectomy (TUR-P) using a photometric method. Seventeen patients were randomly allocated to epidural and 19 patients to general anaesthesia. No significant differences were found between the two groups in the total blood loss peroperatively, the corrected blood loss peroperatively (ml/g/min) or the blood loss postoperatively. The systolic blood pressures were equal in the two groups and no correlation between blood loss and blood pressure was demonstrated. The blood loss per g resected tissue was fairly constant and independent of prostatic weight, but bleeding per minute operating time increased significantly with prostatic size. As the total peroperative blood loss increases with operating time, rapid surgery is a possibility of reducing blood loss in TUR-P. Visual estimation of blood loss during TUR-P of larger glands was unreliable with underestimates of about 100 per cent in one third of the patients, when bleeding exceeded 400 ml. Therefore we recommend an accurate determination of blood loss during resection of larger glands.


Genetic diversity of transmission-blocking vaccine candidate Pvs48/45 in Plasmodium vivax populations in China.
The male gamete fertilization factor P48/45 in malaria parasites is a prime transmission-blocking vaccine (TBV) candidate. Efforts to develop antimalarial vaccines are often thwarted by genetic diversity of the target antigens. Here we evaluated the genetic diversity of Pvs48/45 gene in global Plasmodium vivax populations. We determined 200 Pvs48/45 sequences collected from temperate and subtropical parasite populations in China. Population genetic and evolutionary analyses were performed to determine the levels of genetic diversity, potential signature of selection, and population differentiation. Analysis of the Pvs48/45 sequences from 200 P. vivax parasites collected in a temperate and a tropical region revealed a low level of genetic diversity (π = 0.0012) with 14 single nucleotide polymorphisms, of which 11 were nonsynonymous. Analysis of 344 Pvs48/45 sequences from nine worldwide P. vivax populations detected a total of 38 haplotypes, of which 13 haplotypes were present only once. Multiple tests for selection confirmed a signature of positive selection on Pvs48/45 with selection skewed to the second cysteine domain. Haplotype network analysis and Wright's fixation index showed large geographical differentiation with the presence of continent-or region-specific mutations in this gene. Pvs48/45 displays low levels of genetic diversity with the presence of region-specific mutations. Some of the mutations may be potential epitope targets based on their positions in the predicted structure, highlighting the need for future evaluation of these mutations in designing Pvs48/45-based TBV.


Intravenous Followed by X-ray Fused with MRI-Guided Transendocardial Mesenchymal Stem Cell Injection Improves Contractility Reserve in a Swine Model of Myocardial Infarction.
The aim of this study is to determine the effects of early intravenous (IV) infusion later followed by transendocardial (TE) injection of allogeneic mesenchymal stem cells (MSCs) following myocardial infarction (MI). Twenty-four swine underwent balloon occlusion reperfusion MI and were randomized into 4 groups: IV MSC (or placebo) infusion (post-MI day 2) and TE MSC (or placebo) injection targeting the infarct border with 2D X-ray fluoroscopy fused to 3D magnetic resonance (XFM) co-registration (post-MI day 14). Continuous ECG recording, MRI, and invasive pressure-volume analyses were performed. IV MSC plus TE MSC treated group was superior to other groups for contractility reserve (p = 0.02) and freedom from VT (p = 0.03) but had more lymphocytic foci localized to the peri-infarct region (p = 0.002). No differences were observed in post-MI remodeling parameters. IV followed by XFM targeted TE MSC therapy improves contractility reserve and suppresses VT but does not affect post-MI remodeling and may cause an immune response.


Comparison of proteins in CSF of lateral and IVth ventricles during early development of fetal sheep.
This study examines the relationship between plasma proteins in blood and in CSF in the developing brain of sheep fetuses between 30 and 60 days gestation. Five proteins account for the very high concentration of protein in fetal CSF (over 1000 mg/100 mg/100 ml at 30 days): alpha-feto-protein, fetuin, albumin, alpha 1-antitrypsin and transferrin; the concentration of each protein is similar in lateral and IVth ventricular CSF at 30 days. By 40 days there is considerable decrease in protein concentration in lateral ventricular CSF. At this age in the IVth ventricle the overall total was unchanged, although there were changes in concentration of individual proteins. At 60 days the concentration of each protein in both compartments had fallen below that at 40 days; the marked concentration difference between lateral and IVth ventricular CSF was still present. Experiments using i.v. [125I]- or [3H] labeled plasma proteins in 30-40-day fetuses showed that very little protein penetrated into lateral ventricular CSF by 3-5 h after injection; in the same experiments [125I]albumin reached a CSF/plasma ratio of about 15% in the IVth ventricle (compared with 55% for the natural steady state). Autoradiographic studies carried out on material from the same animals did not give evidence for transfer of labeled protein across the choroid plexuses although any such penetration may have been below the threshold of the method. Other explanations for the high concentration of protein in CSF that were considered include penetration via cerebral vessels and synthesis of plasma proteins by choroid plexus epithelial cells or neurons within the brain.


Effect of coenzyme Q10 on proteomic profile of blood plasma and cytosolic and microsomal fractions of rat hepatocytes during ontogeny.
The proteomic features of blood plasma and subcellular fractions of rat hepatocytes were studied during long-term dietary consumption of coenzyme Q10 as the endogenous mediator of antioxidant and energy homeostasis in the cell. Long-term coenzyme Q10 consumption was followed by the formation of specific nutriproteomes of the microsomal and cytosolic fractions of rat hepatocytes.


Biological removal of pharmaceuticals and personal care products during laboratory soil aquifer treatment simulation with different primary substrate concentrations.
Pharmaceuticals and personal care products (PPCPs) have been detected in bodies of water worldwide, yet their effects on the environment are not fully understood. Recent toxicity studies suggest that mixtures of PPCPs at low concentrations may be detrimental to exposed organisms, highlighting the need to remove PPCPs from wastewater treatment plant effluent before it is discharged to the environment. In this study, the utility of biofilm-based PPCP removal as a means to prevent environmental PPCP contamination was investigated. The removal of 14 PPCPs, each at an initial concentration of 10 μg/L, was studied in laboratory sand columns inoculated with wastewater treatment plant effluent. The examined PPCPs included biosol, biphenylol, p-chloro-m-cresol, p-chloro-m-xylenol, chlorophene, sodium diclofenac, gabapentin, gemfibrozil, 5-fluorouracil, ibuprofen, ketoprofen, naproxen, triclosan, and valproic acid. Ten of the PPCPs were removed by greater than 95% during column passage, while the four other compounds proved more recalcitrant. The effect of the concentration (either 50 or 1000 μg/L) of an easily degradable primary substrate (acetate) supplied along with the mixture of PPCPs was examined. Most of the tested PPCPs were removed consistently by the biofilms regardless of the concentration of acetate, although the extent of removal for three compounds showed dependence on acetate concentration, and two behaved with no reproducible pattern over time. Biofilm protein measurements indicated that the mixture of PPCPs supplied to columns suppressed biofilm growth, suggesting toxicity of the PPCPs to the biofilm communities. This laboratory-scale experiment suggests that biofilm-based water treatment strategies, such as soil aquifer treatment and slow sand filtration, may be well-suited for the removal of many PPCPs from impacted water.


Prevalence of Chlamydia trachomatis in the general population in Germany - a triangulation of data from two population-based health surveys and a laboratory sentinel system.
Chlamydia trachomatis (chlamydia) is a common, frequently asymptomatic, sexually transmitted infection. It can result in severe sequelae, such as ectopic pregnancy and infertility. In Germany, chlamydia is not notifiable. An opportunistic screening program for women < 25 years was introduced in 2008. The aim of this research was to triangulate different data sources to describe the epidemiological situation of chlamydia in Germany and to investigate whether the current target group of the chlamydia screening program aligns with these findings. Urine specimens from participants from population-based health examination surveys of children (2014-17) and adults (2008-11) were tested for chlamydia, using nucleic acid amplification testing. These data were used to generate weighted chlamydia prevalence estimates by age group and sex. Data from a nationwide chlamydia laboratory sentinel system (2014-16) were used to calculate the positive proportion among individuals tested for chlamydia by age, sex and test reason. Using data from the population-based surveys, we found a chlamydia prevalence estimate of 2.8% (95% confidence interval (CI) 1.0-7.5%) among all 15- to 17-year-old girls and of 9.6% (95% CI 0.0-23) among those reporting to be sexually active. In adult women, we found the highest prevalence among 18- to 24-year-olds (all: 2.3%; 95% CI 1.0-5.3%; sexually active: 3.1%; 95% CI 1.3-7.0%). In adult men, we found the highest prevalence among 25- to 29-year-olds (all: 3.5%; 95% CI 1.6-7.7%; sexually active: 3.3%; 95% CI 1.3-7.8%). Data from the chlamydia laboratory sentinel showed the highest positive proportion among those opportunistically screened in 19-year-old women (6.1%; 95%- CI 5.9-6.4%), among those screened due to pregnancy in 15-year-old girls (10%; 95% CI 8.5-12%), and among those tested due to symptoms or a positive partner in 19-year-old women (10%; 95% CI 9.8-11%) and 19-year-old men (24%; 95% CI 22-26%). Chlamydia seems to mainly affect adolescents and young adults in Germany, with similar overall prevalence in men and women, but with slightly different age distributions. Women at highest risk of chlamydia are covered by the current screening program but given the on-going discussions in high-income countries on cost-effectiveness and benefit-to-harm ratio of these programs, the program-aim needs reconsideration.


[Metabolic kinetics and the biological action of 238Pu in the body of rabbits after intratracheal administration].
A study was made of the distribution and biological effect of 238Pu nitrate intratracheally administered to rabbits. The skeleton and liver were the main organs in which 238Pu was secondarily deposited to make 63.5 and 12.9%, respectively, of the total amount administered. For 60 days of observation 15.3% of the amount administered were excreted in feces and urine. With 238Pu dose of 520 kBq/kg acute radiation sickness developed while at a dose of 4 kBq/kg the life span of animals did not vary from the control.


Serum antibody production to botulinum A toxin.
Conflicting data have been reported regarding development of serum antibodies to botulinum A toxin. The purpose of this study is to determine conclusively whether antibody production to this toxin occurs in humans, and, if so, to determine its relationship, if any, to length of treatment, total cumulative dose, and clinical response to treatment. Sixty-five sera samples from 42 adults treated with botulinum A toxin for essential blepharospasm, hemifacial spasm, or spasmodic torticollis were analyzed via a sphere-linked immunodiagnostic assay for antibody production. Results were plotted against length of treatment, number of injections, cumulative dose, and treatment effect produced. Twenty-four (57%) of the 42 patients produced antibodies in all three diagnostic groups. No significant differences were found between antibody producers and nonproducers with respect to age (P = 0.216), length of treatment (P = 0.586), number of injections (P = 0.619), or total cumulative dose (P = 0.286). Within the antibody-producing group, there was no significant correlation between amount of antibody and length of treatment (P = 0.081), number of injections (P = 0.134), or cumulative dose (P = 0.250). The presence of demonstrable antibodies in serum did not affect the clinical responsiveness to injection. Antibody production is present in a majority of patients treated with botulinum A toxin. The sphere-linked immunodiagnostic assay is a reliable and reproducible method for detecting and quantifying these antibodies. When antibody production occurs, it is likely due to variations in individual immune responsiveness and appears to have no direct effect on the patient's clinical response to treatment.


Unusually large isotope effect for the reaction of aromatic amine dehydrogenase. A common feature of quinoproteins?
The reduction of the tryptophan tryptophylquinone [TTQ] prosthetic group of aromatic amine dehydrogenase by dopamine exhibits a deuterium kinetic isotope effect of 8.6-11.7. This effect is partially suppressed in the steady-state. These data support the proposed mechanism for the reductive half-reaction of AADH in which TTQ reduction is linked to proton abstraction from a covalent enzyme-substrate intermediate and in which release of the aldehyde product is partially rate-limiting for the overall reaction. The magnitude of this kinetic isotope effect appears to exceed the semiclassical limit for a proton abstraction. It is also nearly identical to values for the similar reactions catalyzed by methylamine dehydrogenase, which also possesses TTQ, and plasma amine oxidase, which possesses the topaquinone cofactor. These data suggest that these three quinoproteins share a very similar and possible unique mechanism for catalyzing the oxidative deamination of primary amines.