Effect of lecithin:cholesterol acyltransferase on distribution of apolipoprotein A-IV among lipoproteins of human plasma.
The effect of cholesterol esterification on the distribution of apoA-IV in human plasma was investigated. Human plasma was incubated in the presence or absence of the lecithin:cholesterol acyltransferase (LCAT) inhibitor 5,5-dithiobis(2-nitrobenzoic acid) (DTNB) and immediately fractionated by 6% agarose column chromatography. Fractions were monitored for apoA-IV, apoE, and apoA-I by radioimmunoassay (RIA). Incubation resulted in an elevated plasma concentration of cholesteryl ester and in an altered distribution of apoA-IV. After incubation apoA-IV eluted in the ordinarily apoA-IV-poor fractions of plasma that contain small VLDL particles, LDL, and HDL2. Inclusion of DTNB during the incubation resulted in some enlargement of HDL; however, both cholesterol esterification and lipoprotein binding of apoA-IV were inhibited. Addition of DTNB to plasma after incubation and prior to gel filtration had no effect on the apoA-IV distribution when the lipoproteins were immediately fractionated. Fasting plasma apoE was distributed in two or three peaks; in some plasmas there was a small peak that eluted with the column void volume, and, in all plasmas, there were larger peaks that eluted with the VLDL-LDL region and HDL2. Incubation resulted in displacement of HDL apoE to larger lipoproteins and this effect was observed in the presence or absence of DTNB. ApoA-I was distributed in a single broad peak that eluted in the region of HDL and the gel-filtered distribution was unaffected by incubation either in the presence or absence of DTNB. Incubation of plasma that was previously heated to 56 degrees C to inactivate LCAT resulted in no additional movement of apoA-IV onto lipoproteins, unless purified LCAT was present during incubation. The addition of heat-inactivated LCAT to the incubation, had no effect on movement of apoA-IV. These data suggest that human apoA-IV redistribution from the lipoprotein-free fraction to lipoprotein particles appears to be dependent on LCAT action. The mechanism responsible for the increased binding of apoA-IV to the surface of lipoproteins when LCAT acts may involve the generation of "gaps" in the lipoprotein surface due to the consumption of substrate from the surface and additional enlargement of the core. ApoA-IV may bind to these "gaps," where the packing density of the phospholipid head groups is reduced.


Alternative Formulations of Trap Lures for Operational Detection, Population Monitoring, and Outbreak Forecasting of Southern Pine Beetle in the United States.
The southern pine beetle, Dendroctonus frontalis Zimmermann (Coleoptera: Curculionidae: Scolytinae) is a major destructive pest of Pinus L. In the southeastern United States, numbers of this species and a major predator, Thanasimus dubius (Fabricius) (Coleoptera: Cleridae), captured during an annual springtime trapping survey are used to make forecasts of the likelihood and severity of an outbreak during the following summer. We investigated responses by both species to six lure formulations to evaluate their suitability for the survey and allow integration of historical data sets produced with differing lure compositions. Trapping trials were performed at four locations across three states (Louisiana, Mississippi, and Alabama) during spring, and at these and one additional location (North Carolina) in fall 2016. All lures included the pheromone component frontalin. Southern pine beetle preferred lures that additionally included the pheromone component endo-brevicomin and turpentine as a source of host odors (rather than a 7:3 mixture of monoterpenes alpha- and beta-pinene). Thanasimus dubius displayed little discrimination among lure compositions. Lure preferences by southern pine beetle did not differ significantly among locations in spring but were influenced by season. Gas chromatography (GC)-electroantennographic detection analyses with southern pine beetle and GC-mass spectrometry identified numerous known and potential semiochemicals that distinguished volatiles released by the tested host odor devices. The lure combination that included endo-brevicomin and alpha/beta-pinene is recommended for the trapping survey because of its high sensitivity for southern pine beetle and potential for greater data integrity resulting from its reproducible composition.


Analysis of Developing Tooth Germ Innervation Using Microfluidic Co-culture Devices.
Innervation plays a key role in the development, homeostasis and regeneration of organs and tissues. However, the mechanisms underlying these phenomena are not well understood yet. In particular, the role of innervation in tooth development and regeneration is neglected. Several in vivo studies have provided important information about the patterns of innervation of dental tissues during development and repair processes of various animal models. However, most of these approaches are not optimal to highlight the molecular basis of the interactions between nerve fibres and target organs and tissues. Co-cultures constitute a valuable method to investigate and manipulate the interactions between nerve fibres and teeth in a controlled and isolated environment. In the last decades, conventional co-cultures using the same culture medium have been performed for very short periods (e.g., two days) to investigate the attractive or repulsive effects of developing oral and dental tissues on sensory nerve fibres. However, extension of the culture period is required to investigate the effects of innervation on tooth morphogenesis and cytodifferentiation. Microfluidics systems allow co-cultures of neurons and different cell types in their appropriate culture media. We have recently demonstrated that trigeminal ganglia (TG) and teeth are able to survive for a long period of time when co-cultured in microfluidic devices, and that they maintain in these conditions the same innervation pattern that they show in vivo. On this basis, we describe how to isolate and co-culture developing trigeminal ganglia and tooth germs in a microfluidic co-culture system.This protocol describes a simple and flexible way to co-culture ganglia/nerves and target tissues and to study the roles of specific molecules on such interactions in a controlled and isolated environment.


Some 6-aza-5-substituted-2'-deoxyuridines show potent and selective inhibition of herpes simplex virus type 1 thymidine kinase.
The synthesis and X-ray crystal structures of a series of 5-substituted-6-aza-2'-deoxyuridines is reported. These nucleoside analogues inhibit the phosphorylation of thymidine by HSV-1 TK but have no effect on the corresponding human enzyme. Detailed examination of one analogue proves it to be a competitive inhibitor of thymidine with a Ki of 0.34 microM and is a very poor substrate. The analogues are not substrates for the enzyme and also do not inhibit the degradation of thymidine by thymidine phosphorylase. Molecular modelling showed that the inhibitors fit well in the active site of HSV-1 TK, provided the conformation of the sugar moiety is the same for thymidine in the complex.


Phonological facilitation effects on naming latencies and viewing times during noun and verb naming in agrammatic and anomic aphasia.
Phonological priming has been shown to facilitate naming in individuals with aphasia as well as healthy speakers, resulting in faster naming latencies. However, the mechanisms of phonological facilitation (PF) in aphasia remain unclear. Within discrete vs. interactive models of lexical access, this study examined whether PF occurs via the sub-lexical or lexical route during noun and verb naming in agrammatic and anomic aphasia. Thirteen participants with agrammatic aphasia and 10 participants with anomic aphasia and their young and age-matched controls (n=20/each) were tested. Experiment 1 examined noun and verb naming deficit patterns in an off-line confrontation naming task. Experiment 2 examined PF effects on naming both word categories using eyetracking priming paradigm. Results of Experiment 1 showed greater naming difficulty for verbs than for nouns in the agrammatic group, with no difference between the two word categories in the anomic group. For both participant groups, errors were dominated by semantic paraphasias, indicating impaired lexical selection. In the phonological priming task (Experiment 2), young and age-matched control groups showed PF in both noun and verb naming. Interestingly, the agrammatic group showed PF when naming verbs, but not nouns, whereas the anomic group showed PF for nouns only. Consistent with lexically mediated PF in interactive models of lexical access, selective PF for different word categories in our agrammatic and anomic groups suggest that phonological primes facilitate lexical selection via feedback activation, resulting in greater PF for more difficult (i.e., verbs in agrammatic and possibly nouns in anomic group) lexical items.


Sex differences in risk factors for cardiovascular disease: the PERU MIGRANT study.
Although men and women have similar risk factors for cardiovascular disease, many social behaviors in developing countries differ by sex. Rural-to-urban migrants have different cardiovascular risk profiles than rural or urban dwellers. The objective of this study was to evaluate the sex differences with specific cardiovascular risk factors in rural-to-urban migrants. We used the rural-to-urban migrant group of the PERU MIGRANT cross-sectional study to investigate the sex differences in specific cardiovascular risk factors: obesity, hypertension, metabolic syndrome, as well as exposures of socioeconomic status, acculturation surrogates and behavioral characteristics. Logistic regression analysis was used to characterize strength of association between sex and our outcomes adjusting for potential confounders. The sample of migrants was 589 (mean age 46.5 years) and 52.4% were female. In the adjusted models, women were more likely to be obese (OR=5.97; 95%CI: 3.21-11) and have metabolic syndrome (OR=2.22; 95%CI: 1.39-3.55) than men, explaining the greatest variability for obesity and metabolic syndrome but not for hypertension. Our results suggest that interventions for CVD in Peru should be sex-specific and address the unique health needs of migrant populations living in urban shantytowns since the risk factors for obesity and metabolic syndrome differ between males and females.


RAPID3? Aptly named!
The RAPID3 score is the sum of three 0-10 patient self-report scores: pain, functional impairment on MDHAQ, and patient global estimate. It requires 5 seconds for scoring and can be used in all rheumatologic conditions, although it has mostly been used in rheumatoid arthritis where cutoffs for low disease activity (<6/30) or high disease activity (>12/30) have been set. A RAPID3 score of ≤ 3/30 with 1 or 0 swollen joints (RAPID3 ≤ 3 + ≤ SJ1) provides remission criteria comparable to Boolean, SDAI, CDAI, and DAS28 remission criteria, in far less time than a formal joint count. RAPID3 performs as well as the DAS28 in separating active drugs from placebos in clinical trials. RAPID3 also predicts subsequent structural disease progression. RAPID3 can be determined at short intervals at home, allowing the determination of the area under the curve of disease activity between two visits and flare detection. However, RAPID3 should not be seen as a substitute for DAS28 and face to face visits in routine care. Monitoring patient status with only self-report information without a rheumatologist's advice (including joints and physical examination, and consideration of imaging and laboratory tests) may indeed be as undesirable for most patients than joint examination without a patient questionnaire. Conversely, combining the RAPID3 and the DAS28 may consist in faster or more sensitive confirmation that a medication is effective. Similarly, better enquiring of most important concerns of patients (pain, functional status and overall opinion on their disorder) should reinforces patients' confidence in their rheumatologist and treatments.


Total Syntheses of Vancomycin.
A total synthesis of the vancomycin aglycon 1 has eluded synthetic chemists for so many years that the preparation of this molecule has become a vendetta for some groups. Finally, and almost simultaneously, research teams led by Evans and by Nicolaou have succeeded. Similarities and differences in their synthetic approaches are highlighted herein.


Hyperexcitability of Sensory Neurons in Fragile X Mouse Model.
Sensory hypersensitivity and somatosensory deficits represent the core symptoms of Fragile X syndrome (FXS). These alterations are believed to arise from changes in cortical sensory processing, while potential deficits in the function of peripheral sensory neurons residing in dorsal root ganglia remain unexplored. We found that peripheral sensory neurons exhibit pronounced hyperexcitability in Fmr1 KO mice, manifested by markedly increased action potential (AP) firing rate and decreased threshold. Unlike excitability changes found in many central neurons, no significant changes were observed in AP rising and falling time, peak potential, amplitude, or duration. Sensory neuron hyperexcitability was caused primarily by increased input resistance, without changes in cell capacitance or resting membrane potential. Analyses of the underlying mechanisms revealed reduced activity of HCN channels and reduced expression of HCN1 and HCN4 in Fmr1 KO compared to WT. A selective HCN channel blocker abolished differences in all measures of sensory neuron excitability between WT and Fmr1 KO neurons. These results reveal a hyperexcitable state of peripheral sensory neurons in Fmr1 KO mice caused by dysfunction of HCN channels. In addition to the intrinsic neuronal dysfunction, the accompanying paper examines deficits in sensory neuron association/communication with their enveloping satellite glial cells, suggesting contributions from both neuronal intrinsic and extrinsic mechanisms to sensory dysfunction in the FXS mouse model.


Analysis of second-harmonic-generation microscopy in a mouse model of ovarian carcinoma.
Second-harmonic-generation (SHG) imaging of mouse ovaries ex vivo was used to detect collagen structure changes accompanying ovarian cancer development. Dosing with 4-vinylcyclohexene diepoxide and 7,12-dimethylbenz[a]anthracene resulted in histologically confirmed cases of normal, benign abnormality, dysplasia, and carcinoma. Parameters for each SHG image were calculated using the Fourier transform matrix and gray-level co-occurrence matrix (GLCM). Cancer versus normal and cancer versus all other diagnoses showed the greatest separation using the parameters derived from power in the highest-frequency region and GLCM energy. Mixed effects models showed that these parameters were significantly different between cancer and normal (P<0.008). Images were classified with a support vector machine, using 25% of the data for training and 75% for testing. Utilizing all images with signal greater than the noise level, cancer versus not-cancer specimens were classified with 81.2% sensitivity and 80.0% specificity, and cancer versus normal specimens were classified with 77.8% sensitivity and 79.3% specificity. Utilizing only images with greater than of 75% of the field of view containing signal improved sensitivity and specificity for cancer versus normal to 81.5% and 81.1%. These results suggest that using SHG to visualize collagen structure in ovaries could help with early cancer detection.


Mirels Scores in Patients Undergoing Prophylactic Stabilization for Femoral Metastatic Bone Disease in the Veterans Administration Healthcare System.
A retrospective review was performed for patients in the Veterans Administration Healthcare System who underwent prophylactic stabilization of the femur for metastatic disease. The goal was to evaluate indications for prophylactic stabilization through Mirels criteria. All veterans who underwent inpatient prophylactic femoral stabilization between October 2010 and September 2015 were identified. Procedure and demographic variables were collected by using chart review. Provider notes and radiographs were reviewed to calculate Mirels scores. Ninety-two patients underwent confirmed prophylactic stabilization for metastatic bone disease. Lung cancer and multiple myeloma accounted for most lesions. The mean Mirels score was 10.3 (range 7 to 12). 3.2% of patients had a score of 7, and 6.5% had a score of 8. Most lesions were peritrochanteric (66%) and lytic (85%). There was more variability in size (mean 2.3), with 15% being under one third of bony width and 38% between one and two-thirds. The mean pain score was 2.5; 73% reported functional pain. Of lytic and peritrochanteric lesions (53% of all cases), 55% were less than two-thirds the cortical width and 31% lacked functional pain. This retrospective study of prophylactically stabilized metastatic lesions revealed that more than 90% of patients had Mirels scores greater than 8, suggesting a substantial risk of pathologic fracture. Over half of all stabilized lesions were peritrochanteric and lytic. These criteria alone achieve a minimum Mirels score of 8; however, one-third of these lacked functional pain. Notably, Mirels' original paper found location and type criteria to be the least predictive of impending fracture. Contrariwise, functional pain was the most accurate predictor. Multiple studies have found poor specificity of the Mirels criteria. The high scores achievable by the location and type criteria may represent an overrepresentation of their contribution to fracture risk. Reconsideration of the relative weights of each criterion warrants further examination.


GAP-43 mediates retinal axon interaction with lateral diencephalon cells during optic tract formation.
GAP-43 is an abundant intracellular growth cone protein that can serve as a PKC substrate and regulate calmodulin availability. In mice with targeted disruption of the GAP-43 gene, retinal ganglion cell (RGC) axons fail to progress normally from the optic chiasm into the optic tracts. The underlying cause is unknown but, in principle, can result from either the disruption of guidance mechanisms that mediate axon exit from the midline chiasm region or defects in growth cone signaling required for entry into the lateral diencephalic wall to form the optic tracts. Results here show that, compared to wild-type RGC axons, GAP-43-deficient axons exhibit reduced growth in the presence of lateral diencephalon cell membranes. Reduced growth is not observed when GAP-43-deficient axons are cultured with optic chiasm, cortical, or dorsal midbrain cells. Lateral diencephalon cell conditioned medium inhibits growth of both wild-type and GAP-43-deficient axons to a similar extent and does not affect GAP-43-deficient axons more so. Removal or transplant replacement of the lateral diencephalon optic tract entry zone in GAP-43-deficient embryo preparations results in robust RGC axon exit from the chiasm. Together these data show that RGC axon exit from the midline region does not require GAP-43 function. Instead, GAP-43 appears to mediate RGC axon interaction with guidance cues in the lateral diencephalic wall, suggesting possible involvement of PKC and calmodulin signaling during optic tract formation.


In Vivo Imaging Tracking and Immune Responses to Nanovaccines Involving Combined Antigen Nanoparticles with a Programmed Delivery.
Combined nanovaccine can generate robust and persistent antigen-specific immune responses. A combined nanovaccine was developed based on antigen-loaded genipin-cross-linked-polyethyleneimine-antigen nanoparticles and in vivo multispectral fluorescence imaging tracked the antigen delivery of combined nanovaccine. The inner layer antigen nanoparticles carried abundant antigens by self-cross-linking for persistent immune response, whereas the outer antigen on the surface of antigen nanoparticles provided the initial antigen exposure. The delivery of combined nanovaccine was tracked dynamically and objectively by the separation of inner genipin cross-linked antigen nanoparticle and the outer fluorescent antigen. The immune responses of the combined nanovaccine were evaluated including antigen-specific CD4+ and CD8+ T-cell responses, IgG antibody level, immunological memory, and CD8+ cytotoxic T lymphocyte responses. The results indicated that the inner and outer antigens of combined vaccine can be tracked in real time with a programmed delivery by the dual fluorescence imaging. The programmed delivery of the inner and outer antigens induced strong immune responses with a combination of a quick delivery and a persistent delivery. With adequate antigen exposure, the dendritic cells were effectively activated and matured, and following T cells were further activated for immune response. Compared with a single nanoparticle formulation, the combined nanovaccine exactly elicited a stronger antigen-specific immune response.


Frequent loss of expression of the cyclin-dependent kinase inhibitor p27 in epithelial ovarian cancer.
p27Kip1 is a member of the Cip1/Kip1 family of cyclin-dependent kinase inhibitors and is a potential tumor suppressor gene. Low levels of p27 are associated with poor prognosis in a variety of tumors, including breast, colon, prostate, and lung carcinomas. In the present study, p27 protein expression was investigated by immunohistochemistry and Western blot analysis in a series of 82 epithelial ovarian tumors [16 classified as low malignant potential (LMP) and 66 classified as primary ovarian adenocarcinomas]. Immunohistochemical analysis revealed frequent loss of p27 expression in primary ovarian adenocarcinomas (33%), with respect to LMP tumors (6%; P = 0.0009). In addition to nuclear staining, cytoplasmic localization of p27 was noted in 45 (55%) of 82 cases. p27 levels inversely correlated with cdk2 kinase activity in a representative subset of tumors. When the clinical outcome of the patients was evaluated in relationship to p27 status, we observed a significant correlation between presence of p27 staining and a longer time to progression (P = 0.032 by log-rank test). These data indicate that loss of p27 is a frequent event in ovarian carcinomas as compared with LMP tumors, suggesting that these tumor types may have different pathogenesis. p27 levels may also represent a useful prognostic marker for predicting disease recurrence in primary ovarian carcinomas.


Mortality in patients treated for pulmonary embolism.
The hospital course of 144 consecutive patients with pulmonary embolism (PE) demonstrated by pulmonary angiography was reviewed to determine the mortality of patients with treated PE. Twelve patients (8%) died of PE, and eight died of causes other than PE; 124 (86%) survived. Pulmonary embolism was the primary cause of death in only four of the 12 patients who died of PE. Pulmonary embolism contributed to the death of eight other patients, each of whom had associated potentially lethal disease, particularly heart disease. The most important factor affecting mortality was shock due to acute right ventricular failure secondary to massive PE (mortality, 32%). Mortality was not related to magnitude of PE per se; the mortality of patients with massive PE without shock (6%) was the same as that for patients with submassive PE (5%. Patients with PE who survive long enough to have the diagnosis established and appropriate prophylactic therapy begun have an excellent prognosis, unless they have associated severe medical disease.


[Mechanisms of acute coronary syndrome].
The pathogenesis of coronary artery disease is characterized by an increased vasoconstriction, activation of platelet vessel wall interactions as well as the invasion of monocytes into the subintima with deposition of lipids as well as proliferation and migration of vascular smooth muscle cells. In the acute coronary syndromes plaque rupture as well as activation of platelets and coagulation as well as coronary vasoconstriction play an important role. Clinically these mechanisms lead to unstable angina and myocardial infarction. For the understanding of acute coronary syndromes, the mechanisms operative in the healthy blood vessel wall are important. Healthy coronary arteries are in a constant state of vasodilatation, platelets as well as the coagulation cascade are inactivated and vascular smooth muscle cells of the media are quiescent. Endothelial mediators such as nitric oxide, prostacyclin and tissue plasminogen activators and other mediators play an important role in this regard. The continuous release of nitric oxide and prostacyclin keeps the coronary circulation in a state of vasodilatation and inhibits platelet vessel wall interaction. Other mediators of the endothelium also inhibit coagulation and migration and proliferation of vascular smooth muscle cells. On the other hand in patients with cardiovascular risk factors and possibly also in those with genetic disposition as well as with aging, these mechanisms are impaired and in turn increased vasoconstrictor responses of coronary arteries, activation of platelet vessel wall interaction, invasion of monocytes into the subintima with lipid storage and proliferation/-migration of vascular smooth muscle cells occur. A cause oriented therapy of coronary artery disease and of acute coronary syndromes in particular must be based on these pathophysiological mechanisms.


Anti-hyperlipidemic activity of Withania coagulans in streptozotocin-induced diabetes: A potent anti-atherosclerotic agent.
Dyslipidemia is one of the most common complications in diabetes mellitus, which increases risk of premature atherosclerosis. Drugs having antihyperlipidemic activity in addition to their hypoglycemic effect in diabetes may be suitable anti-atherosclerotic agents in diabetic patients. The present study was aimed to investigate the anti-atherosclerotic activity of an aqueous extract of Withania coagulans (AWC) in terms of atherogenic index (AI) in normal and streptozotocin (STZ)-induced diabetes. AWC (1,000 mg/kg body weight, BW) was orally administrated in normal and STZ (70 mg/kg)-induced diabetic rats and levels of glucose, total cholesterol (CHL), high density lipoprotein (HDL)-cholesterol and triglyceride (TG) levels in the plasma were analyzed spectrophotometrically. BW was measured and AI was calculated in each group. Results show that after sub-chronic dosing, AWC reduced plasma glucose levels both in normal and diabetic rats, while significantly decreasing plasma levels of CHL, HDL and TG only in STZ-induced diabetic rats. Repeated administration of AWC also significantly decreased AI and prevented weight loss in STZ-induced diabetic animals. Hence, AWC showed anti-hyperlipidemic activity in diabetic rats and was suggested to be a suitable candidate for the treatment of atherosclerosis associated with diabetes.


Complications in vertebroplasty.
Vertebroplasty is one of the minimally invasive surgery that benefit in pain relief from the osteoporotic or malignancy related vertebral compression fractures. However, many literatures reported both asymptomatic and serious complications. The aim of the present study was to summarize, collect data and report the complication ofvertebroplastyfrom our experience at a single institute. Three hundred and twenty five vertebroplasty procedures from 236 patients performed in our institute were retrospectively reviewed. Data of diagnosis, age at the time of procedure were collected. All complications found were reviewed in detail. Commonly performed procedures were at thoracolumbar junction (51.4%). Osteoporosis was the most common cause of fracture. The present study found 88 (27%) complications with 26 (8%) symptomatic patients. Most common complication was cement leakage, which intervertebral disc was the most common site (42.9). Spinal canal leakage was found in 14 cases (20%). Four out of 14 cases had neurological complications and need further managements. Two cases had neurologic complications from needle injury.Adjacent level collapse found in 13 patients (4%) and remote segment collapse occurred in 5 patients (1.5%). Three had progressive kyphosis required later surgical treatment. One asymptomatic cement pulmonary embolism was found in the present study. The complications of vertebroplasty were mostly asymptomatic, but serious complication such as neurologic injury could occur. Vertebroplasty could be considered a quite safe treatment for osteoporotic vertebral fracture. Meticulous technique should be executed during the procedure to avoid the leakage complication.


Effects of metabolic perturbation on plasma homovanillic acid in schizophrenia. Relationship to prefrontal cortex volume.
Several recent hypotheses suggest that deficits in normal prefrontal cortical inhibition of subcortical dopamine activity result in dysregulated dopamine function and may contribute to the pathophysiology of schizophrenia. These effects seem to be more consistently demonstrable during stressful perturbation of the dopamine system, as opposed to during the resting state. We have developed a novel paradigm involving infusion of the glucose analog, 2-deoxyglucose (2DG), to examine the effects of perturbation on the dopamine metabolite, plasma homovanillic acid (HVA), in schizophrenics and healthy controls. Pharmacologic doses of 2DG (50 mg/kg) and placebo were infused in a double-blind manner. 2-Deoxyglucose-induced effects on plasma HVA were related to magnetic resonance imaging-derived prefrontal cortex volumes. Schizophrenic outpatients (N = 18) and healthy volunteers (N = 11) participated in the study. Schizophrenic patients, as compared with controls, had significantly greater 2DG-induced plasma HVA elevations. These effects were observed in subgroups of neuroleptic-free and neuroleptic-treated patients. Other neuroendocrine (plasma cortisol), physiologic (heart rate, diastolic blood pressure, temperature) and behavioral (self-ratings of stress, fatigue) variables were significantly effected by 2DG but did not differeniate schizophrenics and controls suggesting that the effects on plasma HVA may be relatively specific. Magnetic resonance imaging-derived volumes of the prefrontal cortex were significantly and inversely correlated to 2DG-related peak changes in plasma HVA levels in the schizophrenics. These data support the hypothesis that schizophrenia is associated with abnormal regulation of dopamine and that this deficit may be related to reduced frontal cortical inhibitory influences.


[Evaluating efficacy of transcatheter arterial chemo-embolization combined with radiofrequency ablation on patients with hepatocellular carcinoma by 18FDG-PET/CT].
Judging tumor residue of hepatocellular carcinoma (HCC) after treatment of transcatheter arterial chemo-embolization (TACE) combined with radiofrequency ablation (RFA) by computed tomography (CT) scan is difficult; while 18-fluorodeoxyglucose-positron emission tomography/CT ((18)FDG-PET/CT) has some advantages in this aspect. This study was designed to compare the results of (18)FDG-PET/CT and CT scan in judging tumor residue of HCC after treatment of TACE combined with RFA, and to guide following treatment according to the result of (18)FDG-PET/CT. Thirteen HCC patients with 18 lesions, 0.8-16.0 cm in diameter, were treated in Cancer Center of Sun Yat-sen University from Nov. 2002 to Jun. 2003. Of the 13 patients, 12 were naive patients with 15 lesions; 1 was relapsed with 3 lesions 1 year after hepatectomy. The results of CT and (18)FDG-PET/CT of the 13 patients 2-3 weeks after treatment of TACE combined with RFA were compared. If tumor residue was dictated, a further RFA treatment would be applied within 2-3 weeks. Of the 13 HCC patients that received 1 course of TACE combined with RFA, 11 had tumor residues which were conformed by fine needle biopsy and digital substraction angiography (DSA). CT only detected 5 positive cases; however, (18)FDG-PET/CT detected 10 positive cases. Positive rate of CT was 45.4%, and that of (18)FDG-PET/CT was 90.9%. According to the results of (18)FDG-PET/CT of the 11 patients, after the second course of RFA, 10 patients had no tumor residue; 1 patient survived with uncontrolled lesion 6 weeks after treatment. (18)FDG-PET/CT is better than CT in judging tumor residue of HCC after treatment of TACE combined with RFA or surgery, and in guiding further treatment of HCC.


No acetyl group deficit is evident at the onset of exercise at 90% of maximal oxygen uptake in humans.
The existence of an acetyl group deficit at or above 90% of maximal oxygen uptake (VO(2max)) has proved controversial, with contradictory results likely relating to limitations in previous research. The purpose of the present study was to determine whether the "acetyl group deficit" occurs at the start of exercise at 90%VO(2max) in a well-controlled study. Eight male participants (age: 33.6 +/- 2.0 years; VO(2max): 3.60 +/- 0.21 litres . min(-1)) completed two exercise bouts at 90%VO(2max) for 3 min following either 30 min of saline (control) or dichloroacetate (50 mg . kg(-1) body mass) infusion, ending 15 min before exercise. Muscle biopsies were obtained immediately before and after exercise while continuous non-invasive measures of pulmonary oxygen uptake and muscle deoxygenation were made. Muscle pyruvate dehydrogenase activity was significantly higher before exercise following dichloroacetate infusion (control: 2.67 +/- 0.98 vs. dichloroacetate: 17.9 +/- 1.1 mmol acetyl-CoA . min(-1) . mg(-1) protein, P = 0.01) and resulted in higher pre- and post-exercise muscle acetylcarnitine (pre-exercise control: 3.3 +/- 0.95 vs. pre-exercise dichloroacetate: 8.0 +/- 0.88 vs. post-exercise control: 11.9 +/- 1.1 vs. post-exercise dichloroacetate: 17.2 +/- 1.1 mmol . kg(-1) dry muscle, P < 0.05). However, substrate-level phosphorylation (control: 125 +/- 20 vs. dichloroacetate: 113 +/- 13 mmol adenosine triphosphate . kg(-1) dry muscle) and VO(2) kinetics (control: 19.2 +/- 2.2 vs. dichloroacetate: 22.8 +/- 2.5 s), were unaltered. Furthermore, dichloroacetate infusion blunted the slow component of VO(2) and muscle deoxygenation and slowed muscle deoxygenation kinetics, possibly by enhancing oxygen delivery during exercise. These data support the hypothesis that the "acetyl group deficit" does not occur at or above 90%VO(2max).


Effects of artificial light at night and predator cues on foraging and predator avoidance in the keystone inshore mollusc Concholepas concholepas.
The growth of Artificial Light At Night (ALAN) is potentially having widespread effects on terrestrial and coastal habitats. In this study we addressed both the individual effects of ALAN, as well as its combined effect with predation risk on the behaviour of Concholepas concholepas, a fishery resource and a keystone species in the southeastern Pacific coast. We measured the influence of ALAN and predation risk on this mollusc's feeding rate, use of refuge for light and crawling out of water behaviour. These behavioural responses were studied using light intensities that mimicked levels that had been recorded in coastal habitat exposed to ALAN. Cues were from two species known to prey on C. concholepas during its early ontogeny: the crab Acanthocyclus hassleri and the seastar Heliaster helianthus. The feeding rates of C. concholepas were 3-4 times higher in darkness and in the absence of predator cues. In contrast, ALAN-exposed C. concholepas showed lower feeding activity and were more likely to be in a refuge than those exposed to control conditions. In the presence of olfactory predator cues, and regardless of light treatment, C. concholepas tended to crawl-out of the waterline. We provide evidence to support the hypothesis that exposure to either ALAN or predation risk can alter the feeding behaviour of C. concholepas. However, predator cue recognition in C. concholepas was not affected by ALAN in situations where ALAN and predator cues were both present: C. concholepas continued to forage when predation risk was low, i.e., in darkness and away from predator cues. Whilst this response means that ALAN may not lead to increased predation mortality in C. concholepas, it will reduce feeding activity in this naturally nocturnal species in the absence of dark refugia. Such results may have implications for the long-term health, productivity and sustainability of this keystone species.


Squalamine Restores the Function of the Enteric Nervous System in Mouse Models of Parkinson's Disease.
Parkinson's disease (PD) is a progressive neurodegenerative disorder thought to be caused by accumulation of α-synuclein (α-syn) within the brain, autonomic nerves, and the enteric nervous system (ENS). Involvement of the ENS in PD often precedes the onset of the classic motor signs of PD by many years at a time when severe constipation represents a major morbidity. Studies conducted in vitro and in vivo, have shown that squalamine, a zwitterionic amphipathic aminosterol, originally isolated from the liver of the dogfish shark, effectively displaces membrane-bound α-syn. Here we explore the electrophysiological effect of squalamine on the gastrointestinal (GI) tract of mouse models of PD engineered to express the highly aggregating A53T human α-syn mutant. GI motility and in vivo response to oral squalamine in PD model mice and controls were assessed using an in vitro tissue motility protocol and via fecal pellet output. Vagal afferent response to squalamine was measured using extracellular mesenteric nerve recordings from the jejunum. Whole cell patch clamp was performed to measure response to squalamine in the myenteric plexus. Squalamine effectively restores disordered colonic motility in vivo and within minutes of local application to the bowel. We show that topical squalamine exposure to intrinsic primary afferent neurons (IPANs) of the ENS rapidly restores excitability. These observations may help to explain how squalamine may promote gut propulsive activity through local effects on IPANs in the ENS, and further support its possible utility in the treatment of constipation in patients with PD.


Ovarian function and reproductive hormone levels in girls with Prader-Willi syndrome: a longitudinal study.
The etiology of hypogonadism in girls with Prader-Willi syndrome (PWS) remains uncertain. The aim of the study was to evaluate gonadal function longitudinally in girls and female adolescents with PWS. We performed a longitudinal assessment of anti-müllerian hormone (AMH), gonadotropins, estradiol (E(2)), inhibin B and A, and pubertal development in girls and female adolescents with PWS. Sixty-one girls participating in the Dutch PWS Cohort study participated in the study. Serum AMH, gonadotropins, E(2), and inhibin B and A levels were compared with reference values. AMH levels in girls and female adolescents with PWS were comparable to reference levels between 6 months and 22 yr of age. From 10 yr of age, FSH and LH levels increased to above the 5th percentile compared to reference levels. E(2) and inhibin B levels were in the low normal range in the majority, and inhibin A levels were low but detectable in almost half the female adolescents with PWS. The median age at puberty onset was comparable, but the median ages at attaining Tanner M3 (P = 0.05) and M4 (P < 0.0001) were significantly higher in girls with PWS than in healthy references. Our study shows that the primordial follicle pool and number of small antral follicles are conserved in girls and female adolescents with PWS. We found no classical hypogonadotropic hypogonadism. However, maturation of follicles and progression of pubertal development are impaired, which might be due to dysregulation of LH secretion. Because these impairments are not absolute, ovulation and thus conception cannot be ruled out in individual female adolescents with PWS.


A distinct population of Saccharomyces cerevisiae in New Zealand: evidence for local dispersal by insects and human-aided global dispersal in oak barrels.
Humans have used S. cerevisiae to make alcoholic beverages for at least 5000 years and now this super-model research organism is central to advances in our biological understanding. Current models for S. cerevisiae suggest that its population comprises distinct domesticated and natural groups as well as mosaic strains, but we generally know little of the forces which shape its population structure. In order to test the roles that ecology and geography play in shaping the S. cerevisiae species we examined nine variable microsatellite loci in 172 strains of S. cerevisiae isolated from two spontaneous grape juice ferments, soil, flowers, apiaries and bark in New Zealand. Bayesian analysis shows that the S. cerevisiae in NZ comprise a subdivided but interbreeding population that out-crosses approximately 20% of the time. Some strains contributing to spontaneous ferments cluster with NZ soil/bark isolates, but others cluster with isolates from French oak barrels. It seems some strains have been globally dispersed by humans in oak barrels while some are locally vectored by insects. These data suggest geography is more important than ecology in shaping S. cerevisiae's population structure.


Understanding links between adolescent health and educational attainment.
The educational and economic consequences of poor health during childhood and adolescence have become increasingly clear, with a resurgence of evidence leading researchers to reconsider the potentially significant contribution of early-life health to population welfare both within and across generations. Meaningful relationships between early-life health and educational attainment raise important questions about how health may influence educational success in young adulthood and beyond, as well as for whom its influence is strongest. Using data from the National Longitudinal Survey of Youth 1997, I examine how adolescents'health and social status act together to create educational disparities in young adulthood, focusing on two questions in particular. First, does the link between adolescent health and educational attainment vary across socioeconomic and racial/ethnic groups? Second, what academic factors explain the connection between adolescent health and educational attainment? The findings suggest that poorer health in adolescence is strongly negatively related to educational attainment, net of both observed confounders and unobserved, time-invariant characteristics within households. The reduction in attainment is particularly large for non-Hispanic white adolescents, suggesting that the negative educational consequences of poor health are not limited to only the most socially disadvantaged adolescents. Finally, I find that the link between adolescent health and educational attainment is explained by academic factors related to educational participation and, most importantly, academic performance, rather than by reduced educational expectations. These findings add complexity to our understanding of how the educational consequences of poor health apply across the social hierarchy, as well as why poor health may lead adolescents to complete less schooling.


Tumor-associated mutations in a conserved structural motif alter physical and biochemical properties of human RAD51 recombinase.
Human RAD51 protein catalyzes DNA pairing and strand exchange reactions that are central to homologous recombination and homology-directed DNA repair. Successful recombination/repair requires the formation of a presynaptic filament of RAD51 on ssDNA. Mutations in BRCA2 and other proteins that control RAD51 activity are associated with human cancer. Here we describe a set of mutations associated with human breast tumors that occur in a common structural motif of RAD51. Tumor-associated D149N, R150Q and G151D mutations map to a Schellman loop motif located on the surface of the RecA homology domain of RAD51. All three variants are proficient in DNA strand exchange, but G151D is slightly more sensitive to salt than wild-type (WT). Both G151D and R150Q exhibit markedly lower catalytic efficiency for adenosine triphosphate hydrolysis compared to WT. All three mutations alter the physical properties of RAD51 nucleoprotein filaments, with G151D showing the most dramatic changes. G151D forms mixed nucleoprotein filaments with WT RAD51 that have intermediate properties compared to unmixed filaments. These findings raise the possibility that mutations in RAD51 itself may contribute to genome instability in tumor cells, either directly through changes in recombinase properties, or indirectly through changes in interactions with regulatory proteins.


Self-trapping of the N-H vibrational mode in alpha-helical polypeptides.
Recent calculations on the formation of self-trapped amide group vibrational states in alpha-helical polypeptides [J. Chem. Phys. 124, 134907 (2006)] are extended to the amide N-H normal mode vibrations. First, the adiabatic N-H vibrational single- and two-exciton states are examined by treating the longitudinal chain coordinates as parameters. Then, in using the multiconfiguration time-dependent Hartree method coupled exciton-chain vibrational quantum dynamics are accounted for. Based on the respective exciton-chain vibrational wave function propagation the infrared transient absorption related to a sequential pump-probe experiment is calculated. The modulation of local amide vibrational energies by the longitudinal chain coordinates is found to have a pronounced effect on the broadening of absorption lines. Moreover, the ultrafast exciton transfer in the system is studied in order to characterize the dynamics of the self-trapped single-exciton states on a time scale below 10 ps.


The Impact of Caloric and Non-Caloric Sweeteners on Food Intake and Brain Responses to Food: A Randomized Crossover Controlled Trial in Healthy Humans.
Whether non-nutritive sweetener (NNS) consumption impacts food intake behavior in humans is still unclear. Discrepant sensory and metabolic signals are proposed to mislead brain regulatory centers, in turn promoting maladaptive food choices favoring weight gain. We aimed to assess whether ingestion of sucrose- and NNS-sweetened drinks would differently alter brain responses to food viewing and food intake. Eighteen normal-weight men were studied in a fasted condition and after consumption of a standardized meal accompanied by either a NNS-sweetened (NNS), or a sucrose-sweetened (SUC) drink, or water (WAT). Their brain responses to visual food cues were assessed by means of electroencephalography (EEG) before and 45 min after meal ingestion. Four hours after meal ingestion, spontaneous food intake was monitored during an ad libitum buffet. With WAT, meal intake led to increased neural activity in the dorsal prefrontal cortex and the insula, areas linked to cognitive control and interoception. With SUC, neural activity in the insula increased as well, but decreased in temporal regions linked to food categorization, and remained unchanged in dorsal prefrontal areas. The latter modulations were associated with a significantly lower total energy intake at buffet (mean kcal ± SEM; 791 ± 62) as compared to WAT (942 ± 71) and NNS (917 ± 70). In contrast to WAT and SUC, NNS consumption did not impact activity in the insula, but led to increased neural activity in ventrolateral prefrontal regions linked to the inhibition of reward. Total energy intake at the buffet was not significantly different between WAT and NNS. Our findings highlight the differential impact of caloric and non-caloric sweeteners on subsequent brain responses to visual food cues and energy intake. These variations may reflect an initial stage of adaptation to taste-calorie uncoupling, and could be indicative of longer-term consequences of repeated NNS consumption on food intake behavior.


Neurophysin in the hypothalamo-neurohypophysial system. II. Immunocytochemical studies of the ontogeny of oxytocinergic and vasopressinergic neurons.
Two anti-neurophysin monoclonal antibodies (MABs), PS 36 and PS 41, described in the preceding paper (Ben-Barak, Y, J.T. Russell, M.H. Whitnall, K. Ozato, and H. Gainer (1985) J. Neurosci. 5:000-000), allowed us to specifically stain for oxytocin-associated neurophysin (NP-OT) or vasopressin-associated neurophysin (NP-AVP) in the hypothalamus of developing rats. Staining with these MABs specific for NP-OT or NP-AVP showed that both types of neurophysin appeared in cells in the developing hypothalamus as early as embryonic day (E16) and continued to increase in immunoreactivity throughout fetal life. The literature indicated that oxytocin appears in the system between E20 and E22, much later than vasopressin (E16 to E17), which we confirmed in immunocytochemical experiments using affinity-purified antisera to these hormones. Since the MABs recognize the specific prohormones as well as the specific mature neurophysins (Ben-Barak, Y., J. T. Russell, M.H. Whitnall, K. Ozato, and H. Gainer (1985) J. Neurosci. 5: 81-97), we conclude that there is a developmental delay between the synthesis of the oxytocin prohormone (pro-oxyphysin) and its processing to form oxytocin and NP-OT. The delay in prohormone processing in the oxytocin cells was correlated with a delay in immunocytochemically detectable neurites as compared to the vasopressin cells. This reduced level of axonal and dendritic immunoreactivity was still obvious in the oxytocin cells at 9 days after birth. In contrast, the clustering of cells to form adult-like hypothalamic nuclei appeared to follow similar time courses for the two types of cells. Adult-like distributions of cells staining for NP-OT and NP-AVP were already apparent in the supraoptic and paraventricular nuclei by E17.


The changes in maternal mortality in 1000 counties in mid-Western China by a government-initiated intervention.
Since 2000, the Chinese government has implemented an intervention program to reduce maternal mortality and eliminate neonatal tetanus in accordance with the Millennium Development Goals 5. To assess the effectiveness of this intervention program, we analyzed the level, trend and reasons defining the maternal mortality ratio (MMR) in the 1,000 priority counties before and after implementation of the intervention between 1999 and 2007. The data was obtained from the National Maternal and Child Health Routine Reporting System. The intervention included providing basic and emergency obstetric equipment and supplies to local medical hospitals, and also included providing professional training to local obstetric doctors, development of obstetric emergency centers and "green channel" express referral networks, reducing or waiving the cost of hospital delivery, and conducting community health education. Based on the initiation time of the intervention and the level of poverty, 1,000 counties, containing a total population of 300 million, were categorized into three groups. MMR significantly decreased by about 50%, with an average reduction rate of 9.24%, 16.06%, and 18.61% per year in the three county groups, respectively. The hospital delivery rate significantly increased. Obstetric hemorrhage was the leading cause of maternal deaths and significantly declined, with an average decrease in the MMR of 11.25%, 18.03%, and 24.90% per year, respectively. The magnitude of the MMR, the average reduction rate of the MMR, and the occurrence of the leading causes of death were closely associated with the percentage of poverty. The intervention program implemented by the Chinese government has significantly reduced the MMR in mid-western China, suggesting that well-targeted interventions could be an efficient strategy to reducing MMR in resource-poor areas. Reduction of the MMR not only depends on conducting proven interventions, but also relies on economic development in rural areas with a high burden of maternal death.


Bifidobacterium breve UCC2003 surface exopolysaccharide production is a beneficial trait mediating commensal-host interaction through immune modulation and pathogen protection.
Bifidobacteria constitute a substantial proportion of the human gut microbiota. There are currently many bifidobacterial strains with claimed probiotic attributes. The mechanism through which these strains reside within their host and exert benefits to the host is far from fully understood. We have shown in the case of Bifidobacterium breve UCC2003 that a cell surface exopolysaccharide (EPS) plays a role in in vivo persistence. Biosynthesis of two possible EPSs is controlled by a bidirectional gene cluster which guides alternate EPS synthesis by means of a reorienting promoter. The presence of EPS impacts on host immune response: the wild type, EPS-positive B. breve UCC2003 efficiently evades the adaptive B-cell host response, while its isogenic, EPS-deficient equivalent elicits a strong adaptive immune response. Functionally, EPS positive strains were more resilient to presence of acid and bile and were responsible for reduced colonization levels of Citrobacter rodentium, a gut pathogen. In conclusion, we have found that EPS is important in host interactions and pathogen protection, the latter indicative of a probiotic ability for the EPS of B. breve UCC2003.


A Study of the Reinforcement Effect of MWCNTs onto Polyimide Flat Sheet Membranes.
Polyimides rank among the most heat-resistant polymers and find application in a variety of fields, including transportation, electronics, and membrane technology. The aim of this work is to study the structural, thermal, mechanical, and gas permeation properties of polyimide based nanocomposite membranes in flat sheet configuration. For this purpose, numerous advanced techniques such as atomic force microscopy (AFM), SEM, TEM, TGA, FT-IR, tensile strength, elongation test, and gas permeability measurements were carried out. In particular, BTDA-TDI/MDI (Ρ84) co-polyimide was used as the matrix of the studied membranes, whereas multi-wall carbon nanotubes were employed as filler material at concentrations of up to 5 wt.% All studied films were prepared by the dry-cast process resulting in non-porous films of about 30-50 μm of thickness. An optimum filler concentration of 2 wt.% was estimated. At this concentration, both thermal and mechanical properties of the prepared membranes were improved, and the highest gas permeability values were also obtained. Finally, gas permeability experiments were carried out at 25, 50, and 100 °C with seven different pure gases. The results revealed that the uniform carbon nanotubes dispersion lead to enhanced gas permeation properties.


Allergic cutaneous diseases in hairdressers.
Hairdressers are commonly exposed to water, irritants, sensitizers, and are at risk of occupational skin diseases. Several cases of allergic contact dermatitis and contact urticaria in hairdressers have been described over the last few years. The paper aims to report occupational agents which can induce allergic cutaneous diseases in hairdressers; moreover an analysis of the literature was carried out to review data concerning epidemiology, clinical evaluation, management and prevention. Hairdressing carries a high risk of skin damage, especially for the hands. Allergic contact dermatitis is the most common disorder. Training represents a risk factor, while the role of atopy and nickel sensitivity as etiological agents is controversial. A great number of chemicals may induce allergic cutaneous diseases in hairdressers, such as nickel, thioglycolate, p-phenylendiamine (and other dye-related substances), persulphates, quaternary ammonium compounds, protein hydrolysates, surfactants, cyanoacrylates, latex, rubber haptens and others. Avoidance of further contact with the identified causal sensitizer(s) is the most important measure for management and prevention; to this end, a number of specific preventive strategies should be implemented.


Longitudinal changes in muscle activity during infants' treadmill stepping.
Previous research has described kinetic characteristics of treadmill steps in very stable steppers, in cross-sectional designs. In this study we examined, longitudinally, muscle activation patterns during treadmill stepping, without practice, in 12 healthy infants at 1, 6, and 12 mo of age. We assessed lateral gastrocnemius, tibialis anterior, rectus femoris, and biceps femoris as infants stepped on a treadmill during twelve 20-s trials. Infants showed clear changes in kinematics, such as increased step frequency, increased heel contact at touch down, and more flat-footed contact at midstance. Electromyographic data showed high variability in muscle states (combinations), with high prevalence of all muscles active initially, reducing with age. Agonist-antagonist muscle coactivation also decreased as age increased. Probability analyses showed that across step cycles, the likelihood a muscle was on at any point tended to be <50%; lateral gastrocnemius was the exception, showing an adultlike pattern of probability across ages. In summary, over time, healthy infants produce a wide variety of muscle activation combinations and timings when generating stepping patterns on a treadmill, even if some levels of muscle control arose with time. However, the kinematic stability improved much more clearly than the underlying kinetic strategies. We conclude that although innate control of limb movement improves as infants grow, explore, and acquire functional movement, stepping on a treadmill is a novel and unpracticed one. Hence, developing stable underlying neural activations will only arise as functional practice ensues, similarly to that observed for other functional movements in infancy.


Frequency and Severity of Ear-Nose-Throat (ENT) Symptoms during COVID-19 Infection.
Background and Objectives: Coronavirus disease 2019 (COVID-19) is a new disease entity caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The main symptoms of infection at the onset of the pandemic include dyspnea, cough and high fever. Ear−nose−throat (ENT) symptoms are among the ones presented by patients in the course of infection. The aim of the study was to analyze the frequency of ENT symptoms and to assess their severity and duration in COVID-19 patients. Materials and Methods: The study included 337 patients who had been infected with SARS-CoV-2, as confirmed by a PCR test. The study participants were >18 years old; the mean age was 43.98 years ± 13.47 SD. The convalescents completed a questionnaire that contained 26 questions, including 9 detailed questions related to ENT symptoms, such as sore throat, vertigo, dizziness, hearing disorders, olfactory disorders, taste disturbance, headache, cough and dyspnea. The severity of symptoms was assessed using a Visual Analogue Scale (VAS). Results: The most reported ENT symptoms were olfactory disorders, which occurred in 72% of patients. The second most frequent symptom was taste disturbance (68%), VAS = 6.79 ± 3.01. Vertigo and dizziness were reported by 34% of respondents (VAS = 4.01 ± 2.01). Tinnitus was observed in 15% of patients, VAS = 3.87 ± 1.98; 14% of the subjects reported hearing impairment (VAS = 3.81 ± 2.37). Conclusions: Symptoms related to the sense of smell, taste and hearing are some of the most common symptoms in the course of COVID-19, which is important in the therapeutic and epidemiological management of patients. Delayed diagnosis and treatment of symptoms, especially those related to the hearing organ, may result in greater permanent damage.


Developmental dysregulation of excitatory-to-inhibitory GABA-polarity switch may underlie schizophrenia pathology: A monozygotic-twin discordant case analysis in human iPS cell-derived neurons.
Schizophrenia is a major psychiatric disorder, but the molecular mechanisms leading to its initiation or progression remain unclear. To elucidate the pathophysiology of schizophrenia, we used an in vitro neuronal cell culture model involving human induced pluripotent stem cells (hiPSCs) derived from a monozygotic-twin discordant schizophrenia pair. The cultured neurons differentiated from hiPSCs were composed of a mixture of glutamatergic excitatory neurons and gamma aminobutyric acid (GABA)ergic inhibitory neurons. In the electrophysiological analysis, a different pattern of spontaneous neuronal activity was observed under the condition without any stimulants. The frequency of spontaneous excitatory post-synaptic currents (sEPSCs) was significantly higher in the hiPSC-derived neurons of the patient with schizophrenia than in the control sibling at day-in-vitro 30. However, the synaptic formation was not different between the patient with schizophrenia and the control sibling during the same culture period. To explain underlying mechanisms of higher excitability of presynaptic cells, we focused on the potassium-chloride co-transporter KCC2, which contributes to excitatory-to-inhibitory GABA polarity switch in developing neurons. We also revealed the altered expression pattern of KCC2 in hiPSC-derived neurons from the patient with schizophrenia, which could contribute to understanding the pathology of schizophrenia in the developing nervous system.


Online image-guided intensity-modulated radiotherapy for prostate cancer: How much improvement can we expect? A theoretical assessment of clinical benefits and potential dose escalation by improving precision and accuracy of radiation delivery.
To quantify the theoretical benefit, in terms of improvement in precision and accuracy of treatment delivery and in dose increase, of using online image-guided intensity-modulated radiotherapy (IG-IMRT) performed with onboard cone-beam computed tomography (CT), in an ideal setting of no intrafraction motion/deformation, in the treatment of prostate cancer. Twenty-two prostate cancer patients treated with conventional radiotherapy underwent multiple serial CT scans (median 18 scans per patient) during their treatment. We assumed that these data sets were equivalent to image sets obtainable by an onboard cone-beam CT. Each patient treatment was simulated with conventional IMRT and online IG-IMRT separately. The conventional IMRT plan was generated on the basis of pretreatment CT, with a clinical target volume to planning target volume (CTV-to-PTV) margin of 1 cm, and the online IG-IMRT plan was created before each treatment fraction on the basis of the CT scan of the day, without CTV-to-PTV margin. The inverse planning process was similar for both conventional IMRT and online IG-IMRT. Treatment dose for each organ of interest was quantified, including patient daily setup error and internal organ motion/deformation. We used generalized equivalent uniform dose (EUD) to compare the two approaches. The generalized EUD (percentage) of each organ of interest was scaled relative to the prescription dose at treatment isocenter for evaluation and comparison. On the basis of bladder wall and rectal wall EUD, a dose-escalation coefficient was calculated, representing the potential increment of the treatment dose achievable with online IG-IMRT as compared with conventional IMRT. With respect to radiosensitive tumor, the average EUD for the target (prostate plus seminal vesicles) was 96.8% for conventional IMRT and 98.9% for online IG-IMRT, with standard deviations (SDs) of 5.6% and 0.7%, respectively (p < 0.0001). The average EUDs of bladder wall and rectal wall for conventional IMRT vs. online IG-IMRT were 70.1% vs. 47.3%, and 79.4% vs. 72.2%, respectively. On average, a target dose increase of 13% (SD = 9.7%) can be achieved with online IG-IMRT based on rectal wall EUDs and 53.3% (SD = 15.3%) based on bladder wall EUDs. However, the variation (SD = 9.7%) is fairly large among patients; 27% of patients had only minimal benefit (<5% of dose increment) from online IG-IMRT, and 32% had significant benefit (>15%-41% of dose increment). The ideal maximum dose increment achievable with online IG-IMRT is, on average, 13% with respect to the dose-limiting organ of rectum. However, there is a large interpatient variation, ranging <5%-41%. The results can be applied to calibrate other practical online image-guided techniques for prostate cancer radiotherapy, when intratreatment organ motion/deformation and machine delivery accuracy are considered.


Modification of the Associations Between Duration of Oral Contraceptive Use and Ovarian, Endometrial, Breast, and Colorectal Cancers.
Although oral contraceptive (OC) use is common, the influence of OC use on carcinogenesis is not fully understood. A recent Agency for Healthcare Research and Quality report identified a need to understand the consistency of OC use and cancer associations across subpopulations, including smokers and obese women. To determine whether associations between duration of OC use and risk of specific cancers were modified by lifestyle characteristics. The prospective NIH-AARP Diet and Health Study (enrolled 1995-1996, followed until 2011), with population-based recruitment of AARP members in 6 states and 2 metropolitan areas. All analyses included at least 100 000 women who reported OC use at enrollment. We identified 1241 ovarian, 2337 endometrial, 11 114 breast, and 3507 colorectal cancer cases during follow-up. Data analysis was performed between September 2016 and April 2017. Duration of OC use (never or <1 year [reference], 1-4, 5-9, or ≥10 years). Development of ovarian, endometrial, breast, and colorectal cancers. We examined effect modification by modifiable lifestyle characteristics: cigarette smoking, alcohol consumption, body mass index (BMI), and physical activity. We used Cox models adjusted for age, race, age at menarche, and the modifiers of interest. The analytic population was aged 50 to 71 years (median, 62 years) at enrollment and largely white (91%) and postmenopausal (96%). For ovarian cancer, OC use-associated risk reductions strengthened with duration of use (long-term OC use [≥10 years] HR, 0.60; 95% CI, 0.47-0.76; P < .001 for trend) and were similar across modifiable lifestyle factors. Risk reductions for endometrial cancer strengthened with duration of use (long-term OC use HR, 0.66; 95% CI, 0.56-0.78; P < .001 for trend); the most pronounced reductions were among long-term OC users who were smokers (HR, 0.47; 95% CI, 0.25-0.88), had obese BMIs (0.36; 95% CI, 0.25-0.52), and who exercised rarely (HR, 0.40; 95% CI, 0.29-0.56). Associations between OC use and breast and colorectal cancers were predominantly null. Long-term OC use is consistently associated with reduced ovarian cancer risk across lifestyle factors. We observed the greatest risk reductions for endometrial cancer among women at risk for chronic diseases (ie, smokers, obese BMI). Oral contraceptive use may be beneficial for chemoprevention for a range of women with differing baseline cancer risks.


The Performance-Result Gap in Mixed-Reality Cycling - Evidence From the Virtual Tour de France 2020 on Zwift.
Background: Mixed-reality sports are increasingly reaching the highest level of sport, exemplified by the first Virtual Tour de France, held in 2020. In road races, power output data are only sporadically available, which is why the effect of power output on race results is largely unknown. However, in mixed-reality competitions, measuring and comparing the power output data of all participants is a fundamental prerequisite for evaluating the athlete's performance. Objective: This study investigates the influence of different power output parameters (absolute and relative peak power output) as well as body mass and height on the results in mixed-reality competitions. Methods: We scrape data from all six stages of the 2020 Virtual Tour de France of women and men and analyze it using regression analysis. Third-order polynomial regressions are performed as a cubic relationship between power output and competition result can be assumed. Results: Across all stages, relative power output over the entire distance explains most of the variance in the results, with maximum explanatory power between 77% and 98% for women and between 84% and 99% for men. Thus, power output is the most powerful predictor of success in mixed-reality sports. However, the identified performance-result gap reveals that other determinants have a subordinate role in success. Body mass and height can explain the results only in a few stages. The explanatory power of the determinants considered depends in particular on the stage profile and the progression of the race. Conclusion: By identifying this performance-result gap that needs to be addressed by considering additional factors like competition strategy or the specific use of equipment, important implications for the future of sports science and mixed-reality sports emerge.


Awakening the sleeping giant of urban green in times of crisis-coverage, co-creation and practical guidelines for optimizing biodiversity-friendly and health-promoting residential greenery.
As multiple crises deepen existing inequalities in urban societies within and between neighborhoods, strategically integrating nature-based solutions into the living environment can help reduce negative impacts and improve public health, social cohesion, and well-being. Compared to public green such as parks, semi-public residential greenery is rarely studied, is regularly overlooked by planners, and often receives step-motherly treatment from architects and housing companies. We approximated the area of residential greenery of modernist multi-story apartment complexes in Berlin, Germany. We surveyed residents' suggestions for improving their living environments in vulnerable neighborhoods, report on co-creation experiences, and provide a practical guideline for optimizing health-promoting residential green spaces. The semi-public open space on the doorstep of two-thirds of Berlin's population is highly fragmented and, in total, has a similar area as the public green spaces and a great potential for qualitative development. Just as the suitability of different nature-based solutions to be integrated into the residential greenery depends on building types, resident demands differ between neighborhoods. Residents called for more involvement in design, implementation, and maintenance, frequently proposing that biodiversity-friendly measures be included. As there is no universal solution even for neighborhoods sharing similar structural and socioeconomic parameters, we propose, and have tested, an optimization loop for health-promoting residential greening that involves exploring residents' needs and co-creating local solutions for urban regeneration processes that can be initiated by different actors using bottom-up and/or top-down approaches in order to unlock this potential for healthy, livable and biodiversity friendly cities.


The theory of velocity selective neural recording: a study based on simulation.
This paper describes the improvements to the theory of velocity selective recording and some simulation results. In this method, activity in different groups of axons is discriminated by their propagation velocity. A multi-electrode cuff and an array of amplifiers produce multiple neural signals; if artificial delays are inserted and the signals are added, the activity in axons of the matched velocity are emphasized. We call this intrinsic velocity selective recording. However, simulation shows that interpreting the time signals is then not straight-forward and the selectivity Q(v) is low. New theory shows that bandpass filters improve the selectivity and explains why this is true in the time domain. A simulation study investigates the limits on the available velocity selectivity both with and without additive noise and with reasonable sampling rates and analogue-to-digital conversion parameters. Bandpass filters can improve the selectivity by factors up to 7 but this depends on the speed of the action potential and the signal-to-noise ratio.


Anti-aldosteronergic effect of torasemide.
The diuretic actions of torasemide and furosemide were studied in normotensive rats and in deoxycorticosterone acetate (DOCA)-saline-loaded hypertensive rats. Torasemide (0.3-3 mg/kg) and furosemide (3-30 mg/kg) had a dose-dependent and significant diuretic action in normotensive rats. Potassium retention was only observed in the case of torasemide. Torasemide also had a dose-dependent and significant diuretic action in DOCA-saline-loaded hypertensive rats, whereas furosemide did not. Higher doses of torasemide (10 mg/kg) and furosemide (100 mg/kg) increased both plasma renin activity and aldosterone concentration in normotensive rats in a similar manner. In vivo aldosterone receptor binding was determined to test the possible anti-aldosteronergic effect of torasemide. Torasemide inhibited the binding of aldosterone to its receptor in the cytoplasmic fraction of rat kidney in a dose-dependent manner, while furosemide produced no effect. These results suggest strongly that an anti-aldosteronergic action of torasemide contributes to producing less kaliuresis.


Design, synthesis, and biological evaluation of novel aminopyrimidinylisoindolines as AXL kinase inhibitors.
A novel series of aminopyrimidinylisoindoline derivatives 1a-w having an aminopyrimidine scaffold as a hinge region binding motif were designed and synthesized. Among them, six compounds showed potent inhibitory activities against AXL kinase with IC50 values of submicromolar range. Especially, compound 1u possessing (4-acetylpiperazin-1-yl)phenyl moiety exhibited extremely excellent efficacy (IC50 = <0.00050 μM). Their in vitro antiproliferative activities were tested over five cancer cell lines. Most compounds showed good antiproliferative activities against HeLa cell line. The kinase panel profiling of 50 different kinases and the selected inhibitory activities for the representative compound 1u were carried out. The compound 1u exhibited excellent inhibitory activities (IC50 = <0.00050, 0.025, and 0.050 μM for AXL, MER, and TYRO3, respectively) against TAM family, together with potent antiproliferative activity against MV4-11 cell line (GI50 = 0.10 μM) related to acute myeloid leukemia (AML).


Epidemiology of Fractures Sustained During Electric Scooter Accidents: A Retrospective Review of 563 Cases.
The use of electric scooters (e-scooters) has dramatically increased as they become an attractive alternative for public transportation in busy metropolitan areas worldwide. Despite their benefits, e-scooters challenge the health-care system with poorly understood forms of injuries, mainly orthopaedic fractures. To our knowledge, this study is the first to investigate e-scooter-related orthopaedic fracture patterns, mechanisms of injury, and management. Data on 3,331 e-scooter-related admissions were retrospectively collected between May 2017 and February 2020 in a level-I trauma center. These admissions were analyzed for demographic variables, orthopaedic fracture diagnosis (using the AO/OTA classification), associated injuries, and surgical treatment. During the study period, a total of 716 fractures were diagnosed in 563 patients, with 46.4% of the patients requiring hospitalization. Of 492 upper-limb fractures, 89.2% occurred in a rider fall mechanism; and of 210 lower-limb fractures, 15.7% occurred in rider-vehicle collisions. Fifty-nine percent of long bone fractures were complex fragmentary and/or intra-articular fractures. Orthopaedic surgeons recommended that 225 fractures undergo surgical treatment. The most common upper-limb fracture was AO/OTA class 2R1A, with open reduction and internal fixation of the distal part of the radius being the most common upper-limb procedure (n = 58). The most common lower-limb fracture was AO/OTA class 41C, with open reduction and internal fixation of the proximal part of the tibia being the most common procedure (n = 28). A total of 22 patients (3.9%) required reoperation within 1 year. This investigation provides unique information on demographic characteristics, patterns, and treatment of orthopaedic fractures secondary to the high-energy mechanism of e-scooter injuries. These new in-depth data are important, first, for health-care system preparedness with regard to management and resource allocation to treat these challenging injuries and, second, for legislators promoting safety and injury prevention strategies.


Effect of Hinoki and Meniki Essential Oils on Human Autonomic Nervous System Activity and Mood States.
Meniki (Chamecyparis formosensis) and Hinoki (C. obtusa) are precious conifers with excellent wood properties and distinctive fragrances that make these species popular in Taiwan for construction, interiors and furniture. In the present study, the compositions of essential oils prepared from Meniki and Hinoki were analyzed by gas chromatography-mass spectrometry (GC/MS). Thirty-six compounds were identified from the wood essential oil of Meniki, including Δ-cadinene, γ-cadinene, Δ-cadinol, α-muurolene, calamenene, linalyl acetate and myrtenol; 29 compounds were identified from Hinoki, including α-terpineol, α-pinene, Δ-cadinene, borneol, terpinolene, and limonene. Next, we examined the effect of Meniki and Hinoki essential oils on human autonomic nervous system activity. Sixteen healthy adults received Meniki or Hinoki by inhalation for 5 min, and the physiological and psychological effects were examined. After inhaling Meniki essential oil, participant's systolic blood pressure and heart rate (HR) were decreased, and diastolic blood pressure increased. In addition, sympathetic nervous activity (SNS) was significantly decreased, and parasympathetic activity (PSNS) was significantly increased. On the other hand, after inhaling Hinoki essential oil, systolic blood pressure, heart rate and PSNS were decreased, whereas SNA was increased. Indeed, both Meniki and Hinoki essential oils increased heart rate variability (HRV) in tested adults. Furthermore, in the Profile of Mood States (POMS) test, both Meniki and Hinoki wood essential oils stimulated a pleasant mood status. Our results strongly suggest that Meniki and Hinoki essential oils could be suitable agents for the development of regulators of sympathetic nervous system dysfunctions.


Behavioural traits of colony founders affect the life history of their colonies.
Social arthropods are a major feature in terrestrial ecosystems, and understanding the factors leading to their success is of broad interest. Although many studies have attempted to link colonies' phenotypic composition with their productivity, no study has linked phenotypic composition with the number of offspring colonies formed in the field. I tested whether the behavioural composition of newly founded colonies predicted colony life history patterns in the social spider Anelosimus studiosus. Individual A. studiosus exhibit either an 'aggressive' or 'docile' behavioural type (BT) and BT composition varies among colonies. I constructed artificial colonies of known BT composition and monitored their performance under two conditions: (1) foreign heterospecific spiders present and (2) foreign spiders removed. When heterospecifics were present, colonies founded by docile individuals were invaded by heterospecific spiders more quickly, grew more rapidly in size, produced more offspring colonies per year, but suffered reduced longevity. The life history trade-offs (reproduction, longevity) experienced by colonies resemble those experienced by individuals.


Evaluation of the relationship of serum vitamin D levels in COVID-19 patients with clinical course and prognosis.
SARS-CoV-2 (COVID-19), which emerged in Wuhan, China in December 2019, infected more than six million people in a short time. In COVID-19, the relationship of many laboratory parameters to morbidity and mortality has been defined. In our study, we aimed to determine the relationship of serum vitamin D level to clinical course and prognosis. This study included 108 patients; 88 patients who stayed in Ataturk University and Erzurum City Hospital between March 24, 2020 and May 15, 2020, who were identified as COVID-19 by real-time PCR method from the nasopharyngeal swab and 20 asymptomatic voluntary medical personnel who tested negative for real-time PCR after routine check-up in our hospital. In statistical analysis conducted between healthy control group and vitamin D levels of patients admitted due to COVID-19, it was observed that patients infected with COVID-19 had a lower level (p= 0.004). In 20 patients developing MAS, a lower level of vitamin D was observed (p= 0.004) compared to 68 patients who did not develop. In the comparison of vitamin D levels of the patients (n= 8) who developed exitus in their follow up due to COVID-19, it was observed that vitamin D levels were statistically significantly lower compared to the living (p= 0.009). Due to COVID-19, pandemic, long-running quarantines caused insufficient use of sunlight and worsening of vitamin D deficiency. We wanted to draw attention again with our study to vitamin D which can be responsible for the heavy clinical course of COVID-19 and whose replacement is easy to apply.


SN79, a sigma receptor ligand, blocks methamphetamine-induced microglial activation and cytokine upregulation.
Methamphetamine (METH) abuse is associated with several negative side effects including neurotoxicity in specific brain regions such as the striatum. The precise molecular mechanisms by which METH usage results in neurotoxicity remain to be fully elucidated, with recent evidence implicating the importance of microglial activation and neuroinflammation in damaged brain regions. METH interacts with sigma receptors which are found in glial cells in addition to neurons. Moreover, sigma receptor antagonists have been shown to block METH-induced neurotoxicity in rodents although the cellular mechanisms underlying their neuroprotection remain unknown. The purpose of the current study was to determine if the prototypic sigma receptor antagonist, SN79, mitigates METH-induced microglial activation and associated increases in cytokine expression in a rodent model of METH-induced neurotoxicity. METH increased striatal mRNA and protein levels of cluster of differentiation 68 (CD68), indicative of microglial activation. METH also increased ionized calcium binding adapter molecule 1 (IBA-1) protein expression, further confirming the activation of microglia. Along with microglial activation, METH increased striatal mRNA expression levels of IL-6 family pro-inflammatory cytokines, leukemia inhibitory factor (lif), oncostatin m (osm), and interleukin-6 (il-6). Pretreatment with SN79 reduced METH-induced increases in CD68 and IBA-1 expression, demonstrating its ability to prevent microglial activation. SN79 also attenuated METH-induced mRNA increases in IL-6 pro-inflammatory cytokine family members. The ability of a sigma receptor antagonist to block METH-induced microglial activation and cytokine production provides a novel mechanism through which the neurotoxic effects of METH may be mitigated.


[Pulmonary siderosis: report of a case].
Pulmonary siderosis is one kind of pneumoconiosis caused by the long term inhalation of iron dust. It occurs in a number of occupations including steel rolling and grinding, welding, polishing, casting, boiler scaling, iron ore mining and emery working. Here we report a case of pulmonary siderosis. A 49-year-old male who had worked in an iron foundry for 30 years was admitted because diffuse micronodular lesions were seen in both lung fields on a routine chest radiographic study. A pulmonary function test disclosed a mildly restrictive ventilatory defect. Transbronchial lung biopsy revealed a significant amount of iron dust deposited within a fibrous nodule by which confirmed the diagnosis of pulmonary siderosis.


The Head-off Environmental Asthma in Louisiana (HEAL) study--methods and study population.
In the city of New Orleans, Louisiana, and surrounding parishes (NOLA), children with asthma were perilously impacted by Hurricane Katrina as a result of disrupted health care, high home mold and allergen levels, and high stress. The Head-off Environmental Asthma in Louisiana (HEAL) study was conducted to examine relationships between the post-Katrina environment and childhood asthma in NOLA and assess a novel asthma counselor intervention that provided case management and guidance for reducing home mold and allergen levels. Children (4-12 years old) with moderate-to-severe asthma were recruited from NOLA schools. Over 1 year, they received two clinical evaluations, three home environmental evaluations, and the asthma intervention. Quarterly end points included symptom days, medication use, and unscheduled emergency department or clinic visits. A community advisory group was assembled and informed HEAL at all phases. Of the children (n = 182) enrolled in HEAL, 67% were African American, and 25% came from households with annual incomes < $15,000. HEAL children were symptomatic, averaging 6.6 symptom days in the 2 weeks before baseline, and had frequent unscheduled visits to clinics or emergency departments (76% had at least one unscheduled visit in the preceding 3 months). In this report, we describe study design and baseline characteristics of HEAL children. Despite numerous challenges faced by investigators, study staff, and participants, including destroyed infrastructure, disrupted lives, and lost jobs, HEAL was successful in terms of recruitment and retention, the high quality of data collected that will provide insight into asthma-allergen relationships, and the asthma intervention. This success was attributable to using an adaptive approach and refining processes as needed.


Anxiolytic- and antidepressant-like effects of Silymarin compared to diazepam and fluoxetine in a mouse model of mild traumatic brain injury.
Clinical and experimental studies have shown that mild traumatic brain injury (mTBI) is associated with increased anxiety- and depression-related behaviors and inflammation in the brain. Unfortunately, there are no specific therapies for long-term behavioral consequences of mTBI. This study set out to determine whether silymarin treatment compared to diazepam (DZP) and fluoxetine (FLX) can reduce neuroinflammation, anxiety- and depression-like behaviors after mTBI induction in mice. We used open field, elevated plus maze, light-dark box, zero maze, sucrose preference, forced swim, and tail suspension tests to assess anxiety and depression-like behaviors in mTBI-induced mice. The levels of tumor necrosis factor (TNF)-α protein, a marker of inflammation, in the prefrontal cortex and hippocampus was also measured. This study identified that the long-term treatment with DZP, FLX or SIL results in decreased anxiety and depression-like behaviors in mTBI-induced mice. The results also showed that these drugs reduced TNF-α levels in the prefrontal cortex and hippocampus. In addition, there were no significant differences between the effects of SIL and DZP or SIL and FLX on behavioral and cytokine levels in mTBI-induced mice. Our findings support the idea that mTBI could be a risk factor for anxiety- and depression-related disorders and neuroinflammation in the brain. Taken together, this study demonstrates that DZP, FLX or SIL can significantly reduce anxiety- and depression-like symptoms, and neuroinflammation after mTBI induction in mice.


Definition of the region on NS3 which contains multiple epitopes recognized by dengue virus serotype-cross-reactive and flavivirus-cross-reactive, HLA-DPw2-restricted CD4+ T cell clones.
The epitopes recognized by six CD4+ CD8- cytotoxic T lymphocyte (CTL) clones established from a dengue-3 virus-immune donor were defined. (i) Three CTL clones, JK10, JK34 and JK39, were cross-reactive for dengue virus types 1-4. (ii) One clone, JK28, was cross-reactive for dengue virus types 1-4 and West Nile virus. (iii) Two clones, JK26 and JK49, were cross-reactive for dengue virus types 1-4, West Nile virus and yellow fever virus. The clones, except for JK49, recognized the same epitope on NS3 in an HLA-DPw2-restricted fashion. The smallest synthetic peptide recognized by the five CTL clones was a 10 aa peptide which comprises aa 255-264 on dengue virus NS3. JK49 recognized the overlapping epitope which comprises aa 257-266 in an HLA-DPw2-restricted fashion. Analysis of T cell receptor (TCR) usage by these T cell clones revealed that (i) JK10 and JK34 use V alpha11, and JK34 and JK28 use V beta23, and (ii) the amino acid sequences of the V(D)J junctional region of the TCR were different among these five CTL clones. There were, however, single amino acid conservations among TCRs of some of these T cell clones. These results indicate that the region on NS3 which comprises aa 255-266 contains multiple epitopes recognized by dengue serotype-cross-reactive and flavivirus-cross-reactive CD4+ CTL in an HLA-DPw2-restricted fashion and that a single epitope can be recognized by T cells which have heterogeneous virus specificities.


[Role of alkaline phosphatase in the regulation of the stability of glucose-6-phosphate dehydrogenase from human cells cultivated in vitro].
The causes for different stability of glucose-6-phosphate dehydrogenase in two heteroploid cell strains and in the diploid cell strain of human embryo lungs were investigated. The thermostability of glucose-6-phosphate dehydrogenase was shown to be dependent on the coenzyme (NADP) concentration and to be coupled with the activity of alkaline phosphatase. In diploid and heteroploid cell extracts possessing a low alkaline phosphatase activity glucose-6-phosphate dehydrogenase reveals a high stability. In heteroploid cell extracts having a high activity of alkaline phosphatase a fast hydrolysis of NADP and a decrease of glucose-6-phosphate dehydrogenase stability are observed. Inhibition of alkaline phosphatase by levamisole prior to cell disruption does not increase the stability of glucose-6-phosphate dehydrogenase. Presumably destabilization of glucose-6-phosphate dehydrogenase mediated by alkaline phosphatase occurs in intact cells and is an essential mechanism controlling the enzyme activity.


The Potential of Mesenchymal Stem Cells to Treat Systemic Inflammation in Horses.
One hallmark of mesenchymal stem cells (MSCs) is the ability to differentiate into multiple tissue types which assists in tissue regeneration. Another hallmark of MSCs is their potent anti-inflammatory and immunomodulatory properties and the potential to treat inflammatory, immune-mediated, and ischemic conditions. In equine practice, MSCs have shown efficacy in the treatment of musculoskeletal disorders such as tendinopathy, meniscal tears and cartilage injury. However, there are many equine disease processes and conditions that may benefit from the immunomodulatory properties of MSCs. Examples include conditions associated with overwhelming acute inflammatory response such as systemic inflammatory response syndrome to chronic diseases characterized by a prolonged low level of inflammation such as equine asthma and recurrent uveitis. For the acute inflammatory response processes, there is often high morbidity and mortality with no effective immunomodulatory treatment to prevent the overwhelming synthesis of proinflammatory mediators. For chronic inflammatory disease processes, frequently long-term corticosteroid treatment is the therapeutic mainstay, with serious potential complications. Thus, there is an unmet need for alternative anti-inflammatory treatments for both acute and chronic illnesses in horses. While MSCs show promise for such conditions, much research is needed before a clinically safe and effective treatment will be available. Optimal MSC tissue source, patient vs. donor source (autologous vs. allogeneic) and cell growth conditions need to be determined for each problem. For immediate use, allogeneic MSC treatments is preferable, but immune tolerance and adequate safety require further study. MSC collection and cryopreservation from horses before they are injured or ill, whether from umbilical cord tissue, bone marrow or adipose might become more widespread. Once these fundamental approaches to treating specific diseases with MSCs are determined, the route of administration, dose and timing of administration also need to be studied. To provide a framework for development of MSC immunomodulatory treatments, this article reviews the current understanding of equine MSC anti-inflammatory and immunomodulatory properties and proposes how MSC therapy may be further developed to treat acute onset systemic inflammatory processes and chronic inflammatory diseases.


Evolutionary dynamics of Staphylococcus aureus during progression from carriage to disease.
Whole-genome sequencing offers new insights into the evolution of bacterial pathogens and the etiology of bacterial disease. Staphylococcus aureus is a major cause of bacteria-associated mortality and invasive disease and is carried asymptomatically by 27% of adults. Eighty percent of bacteremias match the carried strain. However, the role of evolutionary change in the pathogen during the progression from carriage to disease is incompletely understood. Here we use high-throughput genome sequencing to discover the genetic changes that accompany the transition from nasal carriage to fatal bloodstream infection in an individual colonized with methicillin-sensitive S. aureus. We found a single, cohesive population exhibiting a repertoire of 30 single-nucleotide polymorphisms and four insertion/deletion variants. Mutations accumulated at a steady rate over a 13-mo period, except for a cluster of mutations preceding the transition to disease. Although bloodstream bacteria differed by just eight mutations from the original nasally carried bacteria, half of those mutations caused truncation of proteins, including a premature stop codon in an AraC-family transcriptional regulator that has been implicated in pathogenicity. Comparison with evolution in two asymptomatic carriers supported the conclusion that clusters of protein-truncating mutations are highly unusual. Our results demonstrate that bacterial diversity in vivo is limited but nonetheless detectable by whole-genome sequencing, enabling the study of evolutionary dynamics within the host. Regulatory or structural changes that occur during carriage may be functionally important for pathogenesis; therefore identifying those changes is a crucial step in understanding the biological causes of invasive bacterial disease.


Thymic macrophages consist of two populations with distinct localization and origin.
Tissue-resident macrophages are essential to protect from pathogen invasion and maintain organ homeostasis. The ability of thymic macrophages to engulf apoptotic thymocytes is well appreciated, but little is known about their ontogeny, maintenance, and diversity. Here, we characterized the surface phenotype and transcriptional profile of these cells and defined their expression signature. Thymic macrophages were most closely related to spleen red pulp macrophages and Kupffer cells and shared the expression of the transcription factor (TF) SpiC with these cells. Single-cell RNA sequencing (scRNA-Seq) showed that the macrophages in the adult thymus are composed of two populations distinguished by the expression of Timd4 and Cx3cr1. Remarkably, Timd4+ cells were located in the cortex, while Cx3cr1+ macrophages were restricted to the medulla and the cortico-medullary junction. Using shield chimeras, transplantation of embryonic thymuses, and genetic fate mapping, we found that the two populations have distinct origins. Timd4+ thymic macrophages are of embryonic origin, while Cx3cr1+ macrophages are derived from adult hematopoietic stem cells. Aging has a profound effect on the macrophages in the thymus. Timd4+ cells underwent gradual attrition, while Cx3cr1+ cells slowly accumulated with age and, in older mice, were the dominant macrophage population in the thymus. Altogether, our work defines the phenotype, origin, and diversity of thymic macrophages.


The evolution of parental care and egg size: a comparative analysis in frogs.
The evolution of parental care and egg size has attracted considerable attention and theoretical debate. Several different hypotheses have been proposed concerning the trajectories of parental care and egg size evolution and the order of specific evolutionary transitions. Few comparative studies have investigated the predictions of these hypotheses. Here, we investigate the evolutionary association between parental care and egg size in frogs in a phylogenetic context. Data on egg size and presence or absence of parental care in various species of frogs was gathered from the scientific literature. As a basis for our comparative analyses, we developed a phylogenetic supertree, by combining the results of multiple phylogenetic analyses in the literature using matrix representation parsimony. Using phylogenetic pairwise comparisons we demonstrated a significant association between the evolution of parental care and large egg size. We then used recently developed maximum likelihood methods to infer the evolutionary order of specific transitions. This analysis revealed that the evolution of large egg size typically precedes the evolution of parental care, rather than the reverse. We discuss the relevance of our results to previous hypotheses concerning the evolution of parental care and egg size.