Mitotane treatment of 32 dogs with cortisol-secreting adrenocortical neoplasms.
Thirty-two dogs with hyperadrenocorticism caused by cortisol-secreting adrenocortical neoplasia were treated with mitotane at an initial daily induction dosage of 27.5 to 75.0 mg/kg of body weight (mean, 46.3 mg/kg) for 10 to 14 days. All dogs received daily maintenance glucocorticoid supplementation during the induction period. After 2 weeks, the ACTH-stimulated serum cortisol concentration had decreased to within or below the reference range for baseline cortisol concentration in 18 (56.3%) of the 32 dogs; the remaining 14 (43.7%) still responded to ACTH administration with serum cortisol concentrations above the reference range. In these 14 dogs, mitotane was continued at a higher daily dosage (mean, 60.7 mg/kg) for an additional 1 to 9 weeks. Serum cortisol concentration subsequently fell within or below the reference range for baseline cortisol concentration in all but 1 dog. In 30 of the 32 dogs, mitotane was continued at an initial mean maintenance dosage of 101.6 mg/kg/wk, divided into 2 to 5 doses. Twenty-two dogs received prednisone daily (0.2 mg/kg) throughout the maintenance period. One or more relapses occurred in 19 (63%) of the 30 dogs. In dogs with relapse, the mean maintenance mitotane dosage was increased from 98.1 mg/kg/wk to a high of 212.4 mg/kg/wk. After a mean maintenance treatment time of 13.2 months, final mean maintenance dosage required in the 30 dogs ranged from 35.3 to 1,273 mg/kg/wk. Adverse effects were seen in 19 (59.4%) of the 32 dogs as a result of a drug toxicosis associated with high-dosage administration of mitotane, low serum cortisol concentration, or both.(ABSTRACT TRUNCATED AT 250 WORDS)


Contrasting enantioselective DNA preference: chiral helical macrocyclic lanthanide complex binding to DNA.
There is great interest in design and synthesis of small molecules which selectively target specific genes to inhibit biological functions in which particular DNA structures participate. Among these studies, chiral recognition has been received much attention because more evidences have shown that conversions of the chirality and diverse conformations of DNA are involved in a series of important life events. Here, we report that a pair of chiral helical macrocyclic lanthanide (III) complexes, (M)-Yb[L(SSSSSS)](3+) and (P)-Yb[L(RRRRRR)](3+), can enantioselectively bind to B-form DNA and show remarkably contrasting effects on GC-rich and AT-rich DNA. Neither of them can influence non-B-form DNA, nor quadruplex DNA stability. Our results clearly show that P-enantiomer stabilizes both poly(dG-dC)(2) and poly(dA-dT)(2) while M-enantiomer stabilizes poly(dA-dT)(2), however, destabilizes poly(dG-dC)(2). To our knowledge, this is the best example of chiral metal compounds with such contrasting preference on GC- and AT-DNA. Ligand selectively stabilizing or destabilizing DNA can interfere with protein-DNA interactions and potentially affect many crucial biological processes, such as DNA replication, transcription and repair. As such, bearing these unique capabilities, the chiral compounds reported here may shed light on the design of novel enantiomers targeting specific DNA with both sequence and conformation preference.


[An elderly case with impending myocardial infarction of endoventricular circular patch plasty with coronary artery bypass grafting].
An 81-year-old man with anterior impending re-myocardial infarction was urgently operated. Preoperative ECG and UCG examinations showed old myocardial infarction and LV aneurysm, respectively. We chose the Dor method (endoventricular circular patch plasty) and CABG aiming at early recovery from congestive heart failure. He fell into respiratory failure temporarily, but otherwise, the clinical course of this operation was good. He was discharged 47 days after the operation on foot. The Dor method combined with CABG is excellent way to relieve patients from congestive heart failure with LV aneurysm.


Fertilization acid of sea urchin eggs is not a consequence of cortical granule exocytosis.
Sea urchin eggs treated with 10 mM NHC1 release a "fertilization acid" although cortical granule exocytosis does not take place. If the eggs are inseminated following ammonia activation, the cortical reaction occurs and a fertilization membrane elevates in the absence of detectable acid release. Examination by electron microscopy of eggs fixed between ammonia activation and insemination confirms the presence of intact granules. Thus, the fertilization acid is not caused by the release of the cortical granules. Ammonia treatment of fertilized eggs stimulates further release of acid. The release of acid can occur repeatedly in ammonia activated eggs if they are washed into normal seawater between successive ammonia treatments. Our results suggest that the release of fertilization acid is related to some metabolic process which can be turned "on or off".


The role of fibroblast growth factor-18 in follicular atresia in cattle.
Although the various members of the fibroblast growth factor (FGF) family are generally mitotic, one member, FGF18, has been shown to increase the rate of apoptosis of ovarian granulosa cells. In the present study, we first determined whether granulosa cells express FGF18 and we then explored the mechanism through which FGF18 increases apoptosis in vitro. Under culture conditions that favored estradiol secretion and CYP19A1 expression, granulosa FGF18 mRNA levels were barely detectable; however, withdrawing gonadotropic support (follicle-stimulating hormone or insulin-like growth factor 1) reduced levels of CYP19A1 mRNA and increased abundance of mRNA encoding the death ligand FASLG and FGF18. Addition of FGF18, but not FGF2, FGF10, or EGF, increased the proportion of apoptotic cells and frequency of caspase 3 activation, and these effects were abrogated by coculture with estradiol. Addition of FGF18 decreased abundance of mRNA encoding the antiapoptotic proteins GADD45B and MDM2, and increased that encoding the proapoptotic protein BBC3; these effects were reversed by coculture with estradiol. The physiological relevance of FGF18 was determined using an in vivo model: injection of FGF18 directly into growing bovine dominant follicles caused cessation of follicle growth by 24 h after injection. Collectively, these data demonstrate that FGF18 is proapoptotic in vivo and may act through a mechanism involving the BBC3-MDM2 pathway.


Chromatographic determination of changes in pigments in spinach (Spinacia oleracea L.) during processing.
The content of individual chlorophyll and carotenoid pigments is determined in three spinach varieties (Lorelei, Springfield, and Ballet) after processing. Raw spinach and spinach that is steam-blanched for 3, 9, or 15 min is stored frozen at -24 degrees C for 6 months. In addition, spinach is air-dried at 75 degrees C, packed in atmospheric air or nitrogen, and stored at ambient temperature for 6 months. Processing has a significant effect on the content of individual chlorophyll and carotenoid pigments; however, there are no differences between varieties in their content of total and individual pigments in raw, frozen spinach. Increasing blanching time resulted in decreased contents of chlorophyll a and b and increased contents of chlorophyll a' and b' and pheophytin a and b because of pheophytinization. Changes in the color because of pheophytinization are only detected after 15 min blanching. The carotenoid pigments are more stable than the chlorophyll pigments during blanching. (all-E)-Violaxanthin is significantly reduced, caused by degradation to other xanthophylls, such as neochrome, during blanching. There are no significant differences in the content of chlorophyll a and b of dried spinach and blanched, frozen spinach. Formation of chlorophyll a' and b', pheophytin a and b, and chlorophyll a-1 and b-1 is observed after drying. The content of pheophytin a and b is significantly lower in dried versus blanched frozen samples. In dried spinach that is stored in atmospheric air, the content of beta-carotene [599 mg/kg dry matter (DM)] is significantly lower compared with nitrogen (766 mg/kg DM), and the content of (all-E)-lutein is lower than in blanched frozen spinach. Neochrome is not detected in raw spinach but in steam-blanched and dried spinach. No differences are observed in the content of (all-E)-neoxanthin, (9'Z)-neoxanthin, (all-E)-violaxanthin, (all-E)-lutein epoxide, or neolutein A and B between spinach that is stored frozen after 3 min blanching and dried spinach.


Building national emission inventories of toxic pollutants in Europe.
The reduction of chemical pollution is a priority in many regional, national, and international policies, including in EU countries. To effectively do so, quantified overviews of pollutant emissions at national levels and with some granularity in their sources, are required. However, current monitoring efforts are often scattered and a quantitative and comprehensive inventory of toxic emissions in Europe is lacking. Toxic pollutants stem from a large variety of emission sources from industry, agriculture, households, etc. and the difficulty to cover all of them is manifest in public databases and official reports, where data gaps across countries and years exist for several substances. Here, we propose a methodology to tackle this problem and build comprehensive and harmonized national inventories of toxic pollutants. Using public databases, official reports, scientific literature and developing extrapolation techniques specific to each emission source, we derived harmonized annual inventories of toxic pollutants in all EU Member States over the years 2000-2014. They present an unprecedented coverage of 805, 572, and 468 substances emitted to air, water and soil, respectively. Although the resulting dataset shows a relatively good agreement with previous inventories of narrower scopes, uncertainties can be identified for specific emission sources and in the development of extrapolation techniques, thus calling for further research in these areas. Such efforts should also explore adaptation of the methodology to derive comprehensive inventories for countries outside EU, where data is scarcer. Nonetheless, the developed national inventories can provide a starting point for territorial chemical footprints of toxic pollutants and could be coupled with environmental impact assessment for gauging the damages to ecosystems and human health from toxic pollutants emitted in Europe. This can ultimately support policy-makers in their pollutants prioritisation and benchmarking across substances and countries towards improved toxic emission reduction policies.


A Newly Designed Fiber-Optic Based Earth Pressure Transducer with Adjustable Measurement Range.
A novel fiber-optic based earth pressure sensor (FPS) with an adjustable measurement range and high sensitivity is developed to measure earth pressures for civil infrastructures. The new FPS combines a cantilever beam with fiber Bragg grating (FBG) sensors and a flexible membrane. Compared with a traditional pressure transducer with a dual diaphragm design, the proposed FPS has a larger measurement range and shows high accuracy. The working principles, parameter design, fabrication methods, and laboratory calibration tests are explained in this paper. A theoretical solution is derived to obtain the relationship between the applied pressure and strain of the FBG sensors. In addition, a finite element model is established to analyze the mechanical behavior of the membrane and the cantilever beam and thereby obtain optimal parameters. The cantilever beam is 40 mm long, 15 mm wide, and 1 mm thick. The whole FPS has a diameter of 100 mm and a thickness of 30 mm. The sensitivity of the FPS is 0.104 kPa/με. In addition, automatic temperature compensation can be achieved. The FPS's sensitivity, physical properties, and response to applied pressure are extensively examined through modeling and experiments. The results show that the proposed FPS has numerous potential applications in soil pressure measurement.


In vivo label-free photoacoustic microscopy of cell nuclei by excitation of DNA and RNA.
Imaging of cell nuclei plays a critical role in cancer diagnosis and prognosis. To image noninvasively cell nuclei in vivo without staining, we developed UV photoacoustic microscopy (UV-PAM), in which 266 nm wavelength UV light excites unlabeled DNA and RNA in cell nuclei to produce photoacoustic waves. We applied UV-PAM to ex vivo imaging of cell nuclei in a mouse lip and a mouse small intestine and to in vivo imaging of the cell nuclei in the mouse skin. The UV-PAM images of unstained cell nuclei match the optical micrographs of the histologically stained cell nuclei. Given intrinsic optical contrast and high spatial resolution, in vivo label-free UV-PAM has potential for unique biological and clinical application.


[Determination of the indices of genetic similarity and diversity using programmable microcomputers].
A complex of programs for microcomputer permitting to compute the similarity and diversity indices of two or few populations, herds, intrabreed types, lines and related groups of animals in automatic operation has been suggested. The absolute values of occurrence of factors and the gene frequencies in polymorphic systems of serum proteins and erythrocyte antigens of blood groups are used in calculation.


Sample Processing and Stability for Urine Biomarker Studies.
Current methods of processing and storing urine samples have not been compared systematically to determine optimal conditions for advancing research on urinary biomarkers. Often, preanalytical handling is nonideal, especially considering the COVID-19 pandemic; consequently, we compared the effects of different short-term storage and processing methods on urinary biomarker measurements. Spot urine samples were collected via a Foley catheter from 20 hospitalized patients from the Yale New Haven Hospital within 48 hours postcardiac surgery. The effects of 3 urine storage and processing methods on biomarkers were tested: (a) 48-hour temporary storage at 4 °C prior to freezing at -80 °C, (b) 48-hour temporary storage at 25 °C prior to freezing at -80 °C, and (c) no centrifugation and immediate storage at -80 °C. Established Meso-Scale Device assay methods were used to measure the urine concentrations of 18 biomarkers: interferon gamma (IFN-ɣ), interleukin (IL)-10, IL-12p70, IL-13, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-18, tumor necrosis factor alpha (TNF-α), epidermal growth factor (EGF), neutrophil gelatinase-associated lipocalin (NGAL), osteopontin (OPN), uromodulin (UMOD), kidney injury molecule-1 (KIM-1), monocyte chemoattractant protein-1 (MCP-1), and chitinase-3-like protein 1 (YKL-40). Measurements of most biomarkers investigated remained stable after temporary storage at 4 °C. IL-6, IL-8, KIM-1, MCP-1, YKL-40, EGF, and NGAL were stable across all 3 processing conditions. IL-12p70 and IL-4 demonstrated significant differences in all tested conditions compared to the reference standard. We identified several notable biomarkers that are robust to variations in preanalytical techniques and can be reliably investigated with nonideal handling conditions.


Point-to-Point Efficient Grafting Improved Compatibility of Boron-Based Fuel-Rich Propellants.
Due to the high mass calorific value, boron powder is widely used in energetic material systems such as solid propellants and ignition powders. Especially, boron powder has very broad application prospects in the field of fuel-rich propellants. However, due to the cross-linking reaction between the boric acid contained on the surface of boron powder and an adhesive called hydroxylated polybutadiene (HTPB), the viscosity of the propellant mixture system increases sharply, which seriously affects the preparation of the propellant. In this paper, "click chemistry" is intended to be used to graft the functional groups on the surface of boron powder to reduce the viscosity of the boron powder and HTPB in the initial mixing stage. In addition, a rheometer was used to test the viscosity of the boron powder and the HTPB system. The test results showed that compared to the viscosity of the raw boron powder system at 24.1 Pa·s, the mixing termination viscosity of the grafted sample was 17.1 Pa·s, a decrease of 29.0%.


Hysteroscopy versus hysterosalpingography in infertile patients.
The purpose of this study is to compare the results of Hysterosalpingography (HSG) and those of Hysteroscopy (H) in order to evaluate intrauterine morphology in a sample of 47 infertile patients. In the HSG sample having no intrauterine pathology, H showed up a pathological condition in 30% of the cases. Similarly, in the cases in which HSG showed pathologies, H gave normal results in 33.3%. Considering H to be a simple diagnostic method, which causes little trauma to the patient, and is more precise than HSG, the authors recommend the hysteroscopic approach, especially in cases of sterility of unknown origin.


Cardiac involvement in the antiphospholipid syndrome.
Antiphospholipid syndrome (APS) is a systemic autoimmune disease, associated with a hypercoagulable state and fetal loss and with other clinical manifestations including cardiac involvement. Cardiac manifestations of APS are valve abnormalities (valve thickening and vegetations), occlusive arterial disease (atherosclerosis and myocardial infarction), intracardiac emboli, ventricular dysfunction, and pulmonary hypertension. Antiphospholipid antibodies (aPLs) may have a role in the accelerated atherosclerotic arterial disease observed in APS, related to their ability to induce endothelial activation. aPLs have been incriminated in the pathogenesis of heart valve lesions in APS patients. Markers of endothelial cell activation are up-regulated with prominent deposition of aPL in heart valves, suggesting aPL deposition initiates an inflammatory process that recruits complement leading to the valve lesion. Autoantibody-mediated endothelial cell activation probably plays a role in sustaining a proadhesive, proinflammatory, and procoagulant phenotype. The heterogeneity of APS clinical manifestations is likely linked to the varied effects that aPL can induce on endothelial cells and to the different functions that endothelial cells display depending on the anatomic localization.


A preliminary study of fluvoxamine maleate on depressive state and serum melatonin levels in patients after cerebral infarction.
Antidepressants have been recommended for the treatment of post-stroke depression (PSD). The purpose of this study was to evaluate the effect of fluvoxamine maleate, a selective serotonin re-uptake inhibitor (SSRI), on depressive state, sleep disturbance, and serum melatonin levels in patients with depressive state after cerebral infarction. Nineteen patients who were hospitalized for cerebral infarction and scored 40 points or higher on the Self Depression Scale (SDS) were enrolled in this study. Nine of the 19 patients received fluvoxamine as a treatment group and the other 10 patients were used as untreated controls. Before and after commencing the drug therapy, the patients were assessed by the SDS, Pittsburgh Sleep Quality Index (PSQI), Japan Stroke Scale for Depression (JSSD), and Japan Stroke Scale for Emotional Disturbance (JSSE), and their serum melatonin levels were measured. The control group underwent the same evaluations as the treatment group. The SDS score improved in the treatment group at 1 week after the start of drug treatment, and in the control group at 1 and 2 weeks into the observation period. In the treatment group, the JSSD and PSQI scores improved and serum melatonin levels increased. The administration of fluvoxamine to patients with depressive state after cerebral infarction alleviated both the depressive state and sleep disturbances. Increased melatonin levels by the administration of fluvoxamine may contribute to improvement in sleep disturbance, one of the major symptoms of depression.


[Blind bifid ureter associated with duplicity and contralateral vesicoureteral reflux].
We report on a pediatric patient who was evaluated for recurrent episodes of urinary tract infection. Evaluation by IVP, cystography and retrograde ureteropyelography revealed a complete ureteral duplicity, grade II vesicoureteral reflux in the lower half of the left kidney and a blind bifid ureter arising at the distal third of the right ureter. This is an uncommon condition (about 150 cases have been reported) difficult to diagnose if undetected by conventional radiological evaluation and retrograde ureteropyelography may be required. It may be associated with urological or other types of malformations. It is generally asymptomatic and with scant consequences, thus treatment is initially conservative. Surgical excision of the blind bifid ureter is warranted only when complications present. In the present case, prophylactic antibiotic therapy was instituted. At one year follow up, the patient is asymptomatic and urine cultures are negative.


Antioxidant and Anti-inflammatory Properties of Resveratrol in Diabetic Nephropathy: A Systematic Review and Meta-analysis of Animal Studies.
Background: Accumulated experimental evidence suggests that resveratrol may have an effect on diabetic nephropathy by inhibiting inflammation and decreasing oxidative stress. However, the credibility of the evidence for this practice is unclear. Thus, we aimed to perform a systematic review and meta-analysis of animal studies to evaluate the antioxidant and anti-inflammatory properties of resveratrol when used in the treatment of diabetic nephropathy. Methods: Electronic bibliographic databases including PubMed, EMBASE, and Web of Science were searched for relevant studies. The methodological quality of animal studies was assessed based on the SYstematic Review Center for Laboratory animal Experimentation Risk of Bias (SYRCLE's RoB) tool. A meta-analysis was performed based on the Cochrane Handbook for Systematic Reviews of Interventions by using RevMan 5.4 software. This study was registered within International Prospective Register of Systematic Reviews (PROSPERO) as number CRD42021293784. Results: Thirty-six qualified studies involving 726 animals were included. There was a significant association of resveratrol with the levels of blood glucose (BG), serum creatinine (Scr), blood urea nitrogen (BUN), catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GPx), and interleukin-1β (IL-1β). Nevertheless, resveratrol treatment did not effectively decrease the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). In addition, more remarkable antioxidant and hypoglycemic effects were observed in type 2 diabetic nephropathy rather than in type 1 diabetic nephropathy based on subgroup analysis. Conclusion: In this meta-analysis, resveratrol can exert its antioxidant activities by reducing the levels of MDA and recovering the activities of SOD, CAT, GSH, and GPx. With regard to pro-inflammatory cytokines, resveratrol had a positive effect on the reduction of IL-1β. However, the analysis indicated that resveratrol had no effect on IL-6 and TNF-α levels, probably because of the methodological quality of the studies and their heterogeneity. Current evidence supports the antioxidant and anti-inflammatory properties of resveratrol, but its relationship with the levels of some inflammatory cytokines such as IL-6 and TNF-α in animals with diabetic nephropathy needs further elucidation.


CHoMP: a chemoenzymatic histology method using clickable probes.
The characterization of aberrant glycosylation patterns in biopsied patient samples represents a remarkable challenge for scientists and medical doctors due to the lack of specific methods for detection. Here, we report the development of a histological method, dubbed CHoMP-chemoenzymatic histology of membrane polysaccharides-for analyzing glycosylation patterns in mammalian tissues. This method exploits a recombinant glycosyltransferase to transfer a monosaccharide analogue equipped with a chemical handle to a specific cell-surface glycan target, which can then be derivatized with imaging probes by using bioorthogonal click chemistry for visualization. We applied CHoMP to survey changes in expression of N-acetyllactosamine (LacNAc) in human samples from patients afflicted with lung adenocarcinoma and observed a sharp decrease in expression levels between normal and early grade tumors, thus suggesting a potential application of this technique in early cancer diagnosis.


Evaluation of five essential oils from aromatic plants of Cameroon for controlling food spoilage and mycotoxin producing fungi.
Five essential oils (EO) extracted from Cymbopogon citratus, Monodora myristica, Ocimum gratissimum, Thymus vulgaris and Zingiber officinale were investigated for their inhibitory effect against three food spoilage and mycotoxin producing fungi, Fusarium moniliforme, Aspergillus flavus and Aspergillus fumigatus. Five strains of each fungus were tested. The agar dilution technique was used to determine the inhibitory effect of each EO on the radial growth of the fungus, and a dose response was recorded. The EO from O. gratissimum, T. vulgaris and C. citratus were the most effective and prevented conidial germination and the growth of all three fungi on corn meal agar at 800, 1000 and 1200 ppm, respectively. Moderate activity was observed for the EO from Z. officinale between 800 and 2500 ppm, while the EO from M. myristica was less inhibitory. These effects against food spoilage and mycotoxin producing fungi indicated the possible ability of each essential oil as a food preservative. A comparative test on the preservative ability of the EO from O. gratissimum and potassium sorbate against A. flavus at pH 3.0 and 4.5 showed that the EO remained stable at both pH, whereas the efficacy of potassium sorbate was reduced at higher pH. We concluded that the EO from O. gratissimum is a potential food preservative with a pH dependent superiority against potassium sorbate, and these are novel scientific information.


Galactosylceramide affects tumorigenic and metastatic properties of breast cancer cells as an anti-apoptotic molecule.
It was recently proposed that UDP-galactose:ceramide galactosyltransferase (UGT8), enzyme responsible for synthesis of galactosylceramide (GalCer), is a significant index of tumor aggressiveness and a potential marker for the prognostic evaluation of lung metastases in breast cancer. To further reveal the role of UGT8 and GalCer in breast cancer progression, tumorigenicity and metastatic potential of control MDA-MB-231 cells (MDA/LUC) and MDA-MB-231 cells (MDA/LUC-shUGT8) with highly decreased expression of UGT8 and GalCer after stable expression of shRNA directed against UGT8 mRNA was studied in vivo in athymic nu/nu mice. Control MDA/LUC cells formed tumors and metastatic colonies much more efficiently in comparison to MDA/LUC-shUGT8 cells with suppressed synthesis of GalCer after their, respectively, orthotopic and intracardiac transplantation. These findings indicate that UGT8 and GalCer have a profound effect on tumorigenic and metastatic properties of breast cancer cells. In accordance with this finding, immunohistochemical staining of tumor specimens revealed that high expression of UGT8 accompanied by accumulation of GalCer in MDA-MB-231 cells is associated with a much higher proliferative index and a lower number of apoptotic cells in comparison to the MDA/LUC-shUGT8 cells. In addition, it was found that expression of UGT8 in MDA-MB-231 cells increased their resistance to apoptosis induced by doxorubicin in vitro. Therefore, these data suggest that accumulation of GalCer in tumor cells inhibits apoptosis, which would facilitates metastatic cells to survive in the hostile microenvironment of tumor in target organ.


Inhibition of leukocyte candidacidal activity by serum from patients with disseminated candidiasis.
The effect of normal serum and serum for seven patients with disseminated candidiasis (DC) on candidacidal activity of normal human neutrophils (PMNL) was studied. PMNL incubated in DC serum had depressed candiacidal activity when compared to studies with normal serum. The candidacidal defect was not related to differences in particle uptake since phagocytic indices were similar with either normal or DC serum. DC serum did not affect PMNL bactericidal activity against Staphylococcus aureus. When Candida albicans (CA) was incubated in either normal or DC sera, washed, and added to PMNL in normal serum, pronounced inhibition of candidacidal activity was seen in all studies with CA previously incubated in DC serum. Dilution of DC serum or absorption of DC serum with heat-killed CA reversed the candidacidal defect. Ultrastructural studies of phagocytized CA demonstrated small ("tight") phagolysosomes and a delay in breakdown of an electron-dense coating on CA previously incubated in DC serum when compared to controls. Inhibition of leukocyte candidacidal activity appeared to be related to high titers of IgG antibodies against CA present in DC serum.


Metal-catalysed oxidation processes in thiosemicarbazones: new complexes with the ligand N-{2-([4-N-ethylthiosemicarbazone]methyl)phenyl}-p-toluenesulfonamide.
The coordination behaviour of a new thiosemicarbazone Schiff-base building block, N-{2-([4-N-ethylthiosemicarbazone]methyl)phenyl}-p-toluenesulfonamide, H2L1 (1), incorporating a bulky tosyl group, towards Mn II, Fe II, Co II, Ni II, Cu II, Zn II, Cd II, Ag I, Sn II, and Pb II has been investigated by means of an electrochemical preparative procedure. Most metal complexes of L1 have the general formula [M(L1)]2.nX (M=Mn, Fe, Co, Ni, Cu, Cd, Pb; n=0-4, X=H2O or CH3CN), as confirmed by the structure of [Pb(L1)]2 (15), in which the lone pair on lead is stereochemically active. This lead(II) complex shows an intense fluorescence emission with a quantum yield of 0.13. In the case of silver, the complex formed was found to possess a stoichiometry of [Ag2(L1)]2.3H2O. During reactions with manganese and copper metals, interesting catalysed processes have been found to take place, with remarkable consequences regarding the ligand skeleton structure. In synthesising the manganese complex, we obtained an unexpected dithiolate thiosemicarbazone tosyl ligand, H2L2, as a side-product, which has been fully characterised, including by X-ray diffraction analysis. In the case of copper, the solid complex has the formula [CuL1]2, but the crystallised product shows the copper atoms coordinated to a new cyclised thiosemicarbazone ligand, H2L3, as in the structures of the complexes [Cu(L3)]2.CH3CN (8) and [Cu(L3)(H2O)]2.CH3CN.H2O (9). The zinc complex [Zn(L1)]4 (12) displays a particular tetranuclear zeolite-type structure capable of hosting small molecules or ions, presumably through hydrogen bonding.


The effects of load on systolic mitral annulus movements by tissue Doppler imaging in cardiac surgery patients.
The main indexes of ventricular function used in clinical practice are strongly influenced by the ventricular load conditions. Tissue Doppler imaging (TDI) has been reported to be a powerful modality that enables assessment of ventricular wall motion with a high temporal and spatial resolution. The study of the systolic movements of the mitral annulus has been proposed as a valid index of myocardial contractility. The aim of this study was to verify if the systolic movements of the mitral annulus by TDI are load-dependent in patients undergoing coronary artery bypass grafting. We evaluated the effect of load changes on systolic (S(M)) and isovolumic (S(IVC)) velocity waves, and on isovolumic acceleration (S(IVA)). Twenty-three patients undergoing coronary artery bypass grafting were evaluate. Load was changed with a rapid infusion of 7 cc/kg of a gelatin solution. The sample volume of TDI was placed at the lateral side of the mitral annulus in the mid-esophageal 4-chamber view. The increase preload resulted in a statistically significant increase of S(M) (8.7+/-1.04 cm/s vs. 9.99+/-0.88 cm/s P=0.01), and a statistically significant increase of S(IVC) (5.65+/-1.95 cm/s vs. 6.75+/-1.53 cm/s P=0.03). S(IVA) does not appear to be changed after the increase of preload (203.45+/-45.69 cm/s(2) vs. 211.41+/-71.28 p=0.81). Only S(IVA) seems to be an independent index of cardiac functionality in cardiac surgery patients and appears to be an optimum index in the clinical evaluation of these patients.


Open-Healing Socket Preservation with a Novel Dense Polytetrafluoroethylene (dPTFE) Membrane: A Retrospective Clinical Study.
Background and objectives: Non-resorbable dense polytetrafluoroethylene (dPTFE) membranes are widely used for regeneration procedures, alone or in combination with particulate materials. The aim of this work was to examine the efficacy of a newly developed dPTFE membrane in the management of extraction socket healing. Materials and Methods: The extraction premolar sockets of 44 patients (20 men and 24 women) were preserved. One group received prosthetic rehabilitation with a fixed partial denture (FPD) (PROS group, N = 19) and a second group received immediate implant placement (IMPL group, N = 25). The PROS group sockets were augmented with a bovine derived xenograft and covered with a newly developed dPTFE membrane prior to FPD rehabilitation. Results: In the IMPL group, socket preservation was combined with immediate implant placement. Before (T0) and 6 months after surgery (T1), horizontal and vertical dimensions were measured with customized stents. No significant differences in alveolar bone loss from T0 to T1 were observed between the PROS and IMPL groups in the horizontal dimension for any tooth type. There was a significant difference in alveolar bone loss from T0 to T1 between the two groups for only single-rooted maxillary premolars in the vertical dimension. Conclusions: The use of the examined new dPTFE membrane consistently led to the preservation of hard tissue in the extraction sites.


Abundance of Early Embryonic Primordial Germ Cells Promotes Zebrafish Female Differentiation as Revealed by Lifetime Labeling of Germline.
Teleost sex differentiation largely depends on the number of undifferentiated germ cells. Here, we describe the generation and characterization of a novel transgenic zebrafish line, Tg(piwil1:egfp-UTRnanos3)ihb327Tg, which specifically labels the whole lifetime of germ cells, i.e., from primordial germ cells (PGCs) at shield stage to the oogonia and early stage of oocytes in the ovary and to the early stage of spermatogonia, spermatocyte, and spermatid in the testis. By using this transgenic line, we carefully observed the numbers of PGCs from early embryonic stage to juvenile stage and the differentiation process of ovary and testis. The numbers of PGCs became variable at as early as 1 day post-fertilization (dpf). Interestingly, the embryos with a high amount of PGCs mainly developed into females and the ones with a low amount of PGCs mainly developed into males. By using transient overexpression and transgenic induction of PGC-specific bucky ball (buc), we further proved that induction of abundant PGCs at embryonic stage promoted later ovary differentiation and female development. Taken together, we generate an ideal transgenic line Tg(piwil1:egfp-UTRnanos3)ihb327Tg which can visualize zebrafish germline for a lifetime, and we have utilized this line to study germ cell development and gonad differentiation of teleost and to demonstrate that the increase of PGC number at embryonic stage promotes female differentiation.


A randomized study of the sitting position for delivery using a newly designed obstetric chair.
A new obstetric chair has been designed to overcome some of the problems of those currently available commercially. The chair has been used to assess the effects of the sitting position in the second stage of labour on the outcome of delivery in 304 women randomly allocated to be delivered either in the chair or in the conventional dorsal position. Delivery in the chair conferred no benefits to mother or baby and resulted in greater mean blood loss and a higher rate of postpartum haemorrhage.


Mu-calpain is functionally required for alpha-processing of Alzheimer's beta-amyloid precursor protein.
Alzheimer's beta-amyloid precursor protein (APP) is normally processed by an unidentified alpha-secretase. A unique feature of this protease is its high sensitivity to phorbol esters, yet the mechanism involved is unclear. We have previously reported that phorbol 12,13-dibutyrate (PDBu) activates calpain, a Ca2+-dependent protease, and PDBu-induced release of APPs (secreted APP) is sensitive to calpain inhibitors, suggesting that calpain is involved in APP alpha-processing. In the present study, we found that PDBu markedly promoted the expression of both mu- and m-calpains in cultured fibroblasts. Dose-response and time course studies revealed that mu-calpain was more sensitive to PDBu than m-calpain and the temporal course of the mu-calpain change coincides better with that of APPs release. Moreover, the stimulatory effect of PDBu on mu-calpain was selectively blocked by mu-calpain-specific siRNA (small interference RNA) and the blockage was accompanied by a concomitant decrease in APPs release. In contrast, m-calpain siRNA did not affect APPs release significantly. Measurement of amyloid beta protein (Abeta) release in the mu-calpain siRNA-treated cells indicated that Abeta40 and Abeta42 levels inversely changed in relation to APPs, and the changes in Abeta42 were more prominent than in Abeta40. Together, these data suggest that calpain, particularly mu-calpain, is a potential candidate for alpha-secretase in the regulated APP alpha-processing, and that changes in this protease can affect the outcome of the overall APP processing.


C-X-C chemokine receptor type 7 antibody enhances neural plasticity after ischemic stroke.
Stromal cell-derived factor-1 and its receptor C-X-C chemokine receptor 4 (CXCR4) have been shown to regulate neural regeneration after stroke. However, whether stromal cell-derived factor-1 receptor CXCR7, which is widely distributed in the developing and adult central nervous system, participates in neural regeneration remains poorly understood. In this study, we established rat models of focal cerebral ischemia by injecting endothelin-1 into the cerebral cortex and striatum. Starting on day 7 after injury, CXCR7-neutralizing antibody was injected into the lateral ventricle using a micro drug delivery system for 6 consecutive days. Our results showed that CXCR7-neutralizing antibody increased the total length and number of sprouting corticospinal tract fibers in rats with cerebral ischemia, increased the expression of vesicular glutamate transporter 1 and growth-related protein 43, markers of the denervated spinal cord synapses, and promoted the differentiation and maturation of oligodendrocyte progenitor cells in the striatum. In addition, CXCR7 antibody increased the expression of CXCR4 in the striatum, increased the protein expression of RAS and ERK1/2 associated with the RAS/ERK signaling pathway, and improved rat motor function. These findings suggest that CXCR7 improved neural functional recovery after ischemic stroke by promoting axonal regeneration, synaptogenesis, and myelin regeneration, which may be achieved by activation of CXCR4 and the RAS/ERK1/2 signaling pathway.


Role of protein kinase C in expression of granulocyte-colony stimulating factor and granulocyte macrophage-colony stimulating factor in lung cancer cells.
We examine the role of protein kinase C (PKC) pathways in the constitutive expression of granulocyte-colony stimulating factor (G-CSF) and granulocyte macrophage-colony stimulating factor (GM-CSF) in lung cancer cells. Two cell lines, OKa-C-1 and MI-4, constitutively produce an abundant dose of G-CSF and GM-CSF. The PKC activator phorbol 12-myristate 13-acetate (PMA) stimulated the production of GM-CSF in a dose-dependent manner and reduced G-CSF in the cell lines. The PKC inhibitor staurosporine had effects opposite to those of PMA in the cell lines. Another PKC activator (4beta-phorbol 12, 13-dibutyrate) and six specific PKC inhibitors (bisindolylmaleimide I, calphostin C, chelerythrine chloride, Gö 6976, PKC inhibitor 19-27, and Ro-32-0432) also worked as well as PMA and staurosporine, respectively. The induction of GM-CSF expression via PKC activation was mediated by the activation of nuclear factor-kappaB. The induction of G-CSF expression via PKC inhibition was mediated by p44/42 mitogen-activated protein kinase and c-Jun N-terminal kinase pathway signaling. GM-CSF may accelerate cell growth and inhibit cell death via PKC activation in the cell lines. G-CSF also seems to reverse growth suppression and cell death induced by PKC inhibition.


Normal ipsilateral/contralateral asymmetries in infant multiple auditory steady-state responses to air- and bone-conduction stimuli.
Two-channel recordings of infants' air- and bone-conduction auditory brainstem responses to brief tones show ipsilateral and contralateral (to the stimulated ear) asymmetries which may be used to isolate which cochlea is the primary contributor to the response. The objective of this study was to determine whether similar ipsilateral/contralateral asymmetries are also present in the air- and bone-conduction "brainstem" (77 to 101 Hz) auditory steady-state responses (ASSRs) of infants. Two-channel ASSRs were recorded in infants (2 to 11 mo) and adults (18 to 40 yr) with normal hearing. Multiple stimuli (carrier frequencies: 500 to 4000 Hz; amplitude/frequency modulated) were presented using a B-71 oscillator on the temporal bone or an ER3-A insert earphone. Bone-conduction ASSR amplitudes, phase delays, and thresholds were obtained for the electroencephalographic (EEG) channels ipsilateral and contralateral to the oscillator temporal-bone placement. Bone-conduction ASSRs were also obtained to the stimulus presented to the opposite temporal bone (at 40 dB HL only). Air-conduction ASSR amplitudes and phase delays were obtained at 60 dB HL in each ear for the EEG channels ipsilateral and contralateral to the transducer. Infants showed more ipsilateral/contralateral asymmetries in both air- and bone-conduction ASSRs compared with adults. Mean bone-conduction ASSR thresholds in infants were 13 to 15 dB higher (i.e., poorer) in the contralateral EEG channel compared with the ipsilateral EEG channel for 500 to 4000 Hz. In adults, there were no large differences (i.e., within 1 dB) between ipsilateral and contralateral ASSR thresholds. Based on ipsilateral/ contralateral threshold differences in infants, interaural attenuation for bone-conducted stimuli was estimated to be at least 10 to 30 dB for most infants. In contrast, most adults showed little interaural attenuation for bone-conducted stimuli. ASSR amplitudes are larger and phase delays are shorter in the ipsilateral EEG channel. For infants, the difference in air-conduction ASSR amplitude between EEG channels was twice that observed for adults. Infants also had greater ASSR amplitude differences between EEG channels for bone-conduction stimuli compared with adults, but the difference was less than that seen for air-conduction stimuli. For air-conduction stimuli, infants had significantly longer phase delays in the contralateral EEG channel compared with the ipsilateral EEG channel. Adults showed no significant differences in air-conduction ASSR phase delay between EEG channels. For bone-conduction stimuli, both infants and adults had significantly longer phase delays in the contralateral EEG channel compared with the ipsilateral EEG channel; the differences in ASSR phase delays between EEG channels were much smaller in infants compared with adults and fewer adults had absent responses in the contralateral EEG channels compared with infants (12% versus 34%). When the transducers were switched to the opposite ear/mastoid, the infant and adult ipsilateral/contralateral asymmetries also switched. Ipsilateral/contralateral asymmetries in air- and bone-conduction ASSRs are clearly present more often and are larger in infants compared with adults. Our findings also suggest that most infants have at least 10 to 30 dB of interaural attenuation to bone-conducted stimuli. These asymmetries in the bone-conduction ASSR have potential as a clinical tool for isolating the cochlea that is contributing to the response in infants.


PD-L1 diagnostics in the neoadjuvant setting: implications of intratumoral heterogeneity of PD-L1 expression in triple negative breast cancer for assessment in small biopsies.
PD-L1 expression is a predictive biomarker for anti-PD-L1 immunotherapy in triple negative breast cancer (TNBC). In the neoadjuvant setting, immunohistochemical (IHC) evaluation of PD-L1 expression can only be performed on small tissue biopsies. In our study we investigated heterogeneity of PD-L1 expression in TNBC, and how reliably PD-L1 expression in small tissue samples reflects PD-L1 expression in larger tumor sections in TNBC. Tissue microarrays (TMAs) were constructed from surgical specimens of 110 patients with TNBC. TMAs contained 4 cores (1 mm in diameter) per patient. To evaluate PD-L1 expression, TMAs were stained with PD-L1 IHC 22C3 PharmDx. Single-core PD-L1 expression was compared to overall PD-L1 expression of each patient's tumor, to ascertain how often small samples of tumor tissue show the same PD-L1 expression as larger tumor samples. Our study found substantial heterogeneity of PD-L1 expression between different TMA cores from the same patient. Heterogeneity was greater in immune cells (ICs) than in tumor cells, in large part due to the uneven distribution of ICs in the tumor. For IC PD-L1 expression, we found that sensitivity can be as low as 0.81 for detecting PD-L1 expression at the 1% threshold most commonly used in breast cancer. Negative predictive value for ICs was 0.7. There is substantial heterogeneity of PD-L1 expression between small tissue samples from the same TNBC tumor, especially for IC expression. This poses challenges for evaluation of PD-L1 expression in the neoadjuvant setting. Negative biopsies should prompt further investigation, and multiple biopsies might be necessary.


Serum Nesfatin-1 Levels in Girls with Idiopathic Central Precocious Puberty.
Nesfatin-1, an anorexigenic neuropeptide, is expressed mainly in the central nervous system and in some peripheral tissues. The role of nesfatin-1 in energy balance has been investigated. Despite the suggestion of a role for nesfatin-1 in reproductive function, data are limited on the role of nesfatin-1 in human puberty. The aim of this study was to investigate the following: i) the role of nesfatin-1 in puberty, and ii) relationship between nesfatin-1 and anthropometric measurements and gonadotropin levels in girls with idiopathic central precocious puberty (CPP). Twenty-four girls with CPP (7.68±1.02 years) and 20 female, prepubertal, healthy controls (7.48±0.88 years) were enrolled in the study. All patients with CPP were treated by the intramuscular administration of leuprolide acetate at a daily dose of 3.75 mg for 28 days. Nesfatin-1 was measured before and during treatment. There was no difference in serum nesfatin-1 levels in girls with CPP and healthy controls [5.67 (2.5-20.6) mmol/L and 5.75 (2.51-9.64) mmol/L], respectively. There was a negative correlation between nesfatin-1 levels and body weight and body mass index-standard deviation score (p=0.01, r=-0.83; p=0.025, r=-0.81, respectively). No correlation was found between nesfatin-1 and gonadotropin, estradiol levels, uterine length or endometrial thickness. The results of this study suggest that there are no differences between girls with CPP and healthy, prepubertal girls regarding nesfatin-1 levels.


Understanding pressurized metered dose inhaler performance.
Deepening the current understanding of the factors governing the performance of the pressurized metered dose inhaler (pMDI) has the potential to benefit patients by providing improved drugs for current indications as well as by enabling new areas of therapy. Although a great deal of work has been conducted to this end, our knowledge of the physical mechanisms that drive pMDI performance remains incomplete. This review focuses on research into the influence of device and formulation variables on pMDI performance metrics. Literature in the areas of dose metering, atomization and aerosol evolution and deposition is covered, with an emphasis on studies of a more fundamental nature. Simple models which may be of use to those developing pMDI products are summarized. Although researchers have had good success utilizing an empirically developed knowledge base to predict pMDI performance, such knowledge may not be applicable when pursuing innovations in device or formulation technology. Developing a better understanding of the underlying mechanisms is a worthwhile investment for those working to enable the next generation of pMDI products.


Neural adaptation to resistance training: changes in evoked V-wave and H-reflex responses.
Combined V-wave and Hoffmann (H) reflex measurements were performed during maximal muscle contraction to examine the neural adaptation mechanisms induced by resistance training. The H-reflex can be used to assess the excitability of spinal alpha-motoneurons, while also reflecting transmission efficiency (i.e., presynaptic inhibition) in Ia afferent synapses. Furthermore, the V-wave reflects the overall magnitude of efferent motor output from the alpha-motoneuron pool because of activation from descending central pathways. Fourteen male subjects participated in 14 wk of resistance training that involved heavy weight-lifting exercises for the muscles of the leg. Evoked V-wave, H-reflex, and maximal M-wave (M(max)) responses were recorded before and after training in the soleus muscle during maximal isometric ramp contractions. Maximal isometric, concentric, and eccentric muscle strength was measured by use of isokinetic dynamometry. V-wave amplitude increased approximately 50% with training (P < 0.01) from 3.19 +/- 0.43 to 4.86 +/- 0.43 mV, or from 0.308 +/- 0.048 to 0.478 +/- 0.034 when expressed relative to M(max) (+/- SE). H-reflex amplitude increased approximately 20% (P < 0.05) from 5.37 +/- 0.41 to 6.24 +/- 0.49 mV, or from 0.514 +/- 0.032 to 0.609 +/- 0.025 when normalized to M(max). In contrast, resting H-reflex amplitude remained unchanged with training (0.503 +/- 0.059 vs. 0.499 +/- 0.063). Likewise, no change occurred in M(max) (10.78 +/- 0.86 vs. 10.21 +/- 0.66 mV). Maximal muscle strength increased 23-30% (P < 0.05). In conclusion, increases in evoked V-wave and H-reflex responses were observed during maximal muscle contraction after resistance training. Collectively, the present data suggest that the increase in motoneuronal output induced by resistance training may comprise both supraspinal and spinal adaptation mechanisms (i.e., increased central motor drive, elevated motoneuron excitability, reduced presynaptic inhibition).


Index of contractile asymmetry improves patient selection for CRT: a proof-of-concept study.
Nearly one-third of heart failure (HF) patients do not respond to cardiac resynchronization therapy (CRT) despite having left bundle branch block (LBBB). The aim of the study was to investigate a novel method of quantifying left ventricular (LV) contractile asymmetry in HF. Patients with HF and LBBB undergoing CRT (n = 89, 37.1% females, 68 ± 9 years, ischemic etiology in 61%, LV ejection fraction 27.1 ± 7.1%) were analyzed. LV longitudinal systolic strain rate values were extracted from curved anatomical M-mode plots of standard long-axis 2D-echocardiography images and cubic spline interpolation was used to generate a 3D-phantom. Index of contractile asymmetry (ICA) was calculated based on standard deviation of differences in strain rate of opposing walls. Average ICA was individually assessed pairwise in 12 opposing 30-degree LV sectors. Reduction in LV end-systolic volume (ESV) ≥15% after 6 months was considered as positive response to CRT. CRT response was found in 66 (74.2%) patients. Responders with both ischemic and non-ischemic cardiomyopathy had a higher and more extensive contractile asymmetry at baseline and achieved a greater ICA reduction after CRT than non-responders. Higher baseline ICA predicted higher degree and wider extent of ICA improvement. Also, both ICA at baseline and reduction of ICA correlated with the degree of ESV reduction after CRT. Quantification of asymmetrical LV activation in 3D by ICA provides valuable insights into LV contraction in case of LBBB and is a promising tool for improved patient selection for CRT.


Application of quantitative models from population biology and evolutionary game theory to tumor therapeutic strategies.
Quantitative models from population biology and evolutionary game theory frame the tumor-host interface as a dynamical microenvironment of competing tumor and normal populations. Through this approach, critical parameters that control the outcome of this competition are identified and the conditions necessary for formation of an invasive cancer are defined. Perturbations in these key parameters that destabilize the cancer solution of the state equations and produce tumor regression can be predicted. The mathematical models demonstrate significant theoretical limitations in therapies based solely on cytotoxic drugs. Because these approaches do not alter critical parameters controlling system dynamics, the tumor population growth term will remain positive as long as any individual cells are present so that the tumor will invariably recur unless all proliferative cells are killed. The models demonstrate that such total effectiveness is rendered unlikely by the genotypic heterogeneity of tumor populations (and, therefore, the variability of their response to such drugs) and the ability of tumor cells to adapt to these proliferation constraints by evolving resistant phenotypes. The mathematical models support therapeutic strategies that simultaneously alter several of the key parameters in the state equations. Furthermore, the models demonstrate that administration of cytotoxic therapies will, by reducing the tumor population density, create system dynamics more conducive to perturbations by biological modifiers.


Truncated tests for combining evidence of summary statistics.
To date, thousands of genetic variants to be associated with numerous human traits and diseases have been identified by genome-wide association studies (GWASs). The GWASs focus on testing the association between single trait and genetic variants. However, the analysis of multiple traits and single nucleotide polymorphisms (SNPs) might reflect physiological process of complex diseases and the corresponding study is called pleiotropy association analysis. Modern day GWASs report only summary statistics instead of individual-level phenotype and genotype data to avoid logistical and privacy issues. Existing methods for combining multiple phenotypes GWAS summary statistics mainly focus on low-dimensional phenotypes while lose power in high-dimensional cases. To overcome this defect, we propose two kinds of truncated tests to combine multiple phenotypes summary statistics. Extensive simulations show that the proposed methods are robust and powerful when the dimension of the phenotypes is high and only part of the phenotypes are associated with the SNPs. We apply the proposed methods to blood cytokines data collected from Finnish population. Results show that the proposed tests can identify additional genetic markers that are missed by single trait analysis.


Quantitation of human serum polymeric IgA, IgA1 and IgA2 immunoglobulin by enzyme immunoassay.
This report concerns the relative quantitation of serum polymeric IgA and polymeric IgA subclass concentrations by enzyme immunoassay (EIA). The assay relies on the specific binding of polymeric IgA to secretory component. Competition between pentameric IgM and polymeric IgA for binding to secretory component was observed. Thus, samples were adsorbed for IgM by affinity chromatography before the EIA was performed. The assay was used to determine an age-related range of serum polymeric IgA concentrations and to compare the polymeric IgA concentrations in patients with IgA nephropathy (n = 50) to those of controls (n = 50). The serum concentrations of both polymeric IgA and polymeric IgA1 increased with age reaching adult values of around 12 years of age. Polymeric IgA2 concentrations did not reach adult levels until 18 years of age. The ratio of the polymeric IgA concentration to the total serum IgA concentration was found to be significantly increased in children under 2 years of age compared with those over 4 years of age (Mann-Whitney U-test, P less than 0.01). Patients with IgA nephropathy had significantly increased concentrations of polymeric IgA (P = 0.001) and polymeric IgA1 (P = 0.001) but similar polymeric IgA2 concentrations to controls.


Nucleotide sequencing and characterization of Pseudomonas putida catR: a positive regulator of the catBC operon is a member of the LysR family.
Pseudomonas putida utilizes the catBC operon for growth on benzoate as a sole carbon source. This operon is positively regulated by the CatR protein, which is encoded from a gene divergently oriented from the catBC operon. The catR gene encodes a 32.2-kilodalton polypeptide that binds to the catBC promoter region in the presence or absence of the inducer cis-cis-muconate, as shown by gel retardation studies. However, the inducer is required for transcriptional activation of the catBC operon. The catR promoter has been localized to a 385-base-pair fragment by using the broad-host-range promoter-probe vector pKT240. This fragment also contains the catBC promoter whose -35 site is separated by only 36 nucleotides from the predicted CatR translational start. Dot blot analysis suggests that CatR binding to this dual promoter-control region, in addition to inducing the catBC operon, may also regulate its own expression. Data from a computer homology search using the predicted amino acid sequence of CatR, deduced from the DNA sequence, showed CatR to be a member of a large class of procaryotic regulatory proteins designated the LysR family. Striking homology was seen between CatR and a putative regulatory protein, TfdS.


Brucellosis status report.
Tremendous progress was made in the eradication of brucellosis for about 15 yr beginning in 1955. However, progress has been little in the last few years. This has been due to inflation and commitment of resources to other programs which had a higher priority at the time. We need additional resources, strengthened program procedures, and a renewed commitment on the part of all people and organizations involved in the eradication effort.


Different strategies of cadmium detoxification in the submerged macrophyte Ceratophyllum demersum L.
The heavy metal cadmium (Cd) is highly toxic to plants. To understand the mechanisms of tolerance and resistance to Cd, we treated the rootless, submerged macrophyte Ceratophyllum demersum L. with sub-micromolar concentrations of Cd under environmentally relevant conditions. X-ray fluorescence measurements revealed changing distribution patterns of Cd and Zn at non-toxic (0.2 nM, 2 nM), moderately toxic (20 nM) and highly toxic (200 nM) levels of Cd. Increasing Cd concentrations led to enhanced sequestration of Cd into non-photosynthetic tissues like epidermis and vein. At toxic Cd concentrations, Zn was redistributed and mainly found in the vein. Cd treatment induced the synthesis of phytochelatins (PCs) in the plants, with a threshold of induction already at 20 nM Cd for PC3. In comparison, in plants treated with Cu, elevated PC levels were detected only at the highest concentrations (100-200 nM Cu). Our results show that also non-accumulators like C. demersum store toxic metals in tissues where the heavy metal interferes least with metabolic pathways, but remaining toxicity interferes with micronutrient distribution. Furthermore, we found that the induction of phytochelatins is not proportional to metal concentration, but has a distinct threshold, specific for each PC species. Finally we could show that 20 nM Cd, which was previously regarded as non-toxic to most plants, already induces detoxifying mechanisms.


Characterization of a novel bile alcohol sulfate released by sexually mature male sea lamprey (Petromyzon marinus).
A sulphate-conjugated bile alcohol, 3,12-diketo-4,6-petromyzonene-24-sulfate (DKPES), was identified using bioassay-guided fractionation from water conditioned with sexually mature male sea lamprey (Petromyzon marinus). The structure and relative stereochemistry of DKPES was established using spectroscopic data. The electro-olfactogram (EOG) response threshold of DKPES was 10(-7) Molar (M) and that of 3-keto petromyzonol sulfate (3 KPZS; a known component of the male sea lamprey sex pheromone) was 10(-10) M. Behavioural studies indicated that DKPES can be detected at low concentrations by attracting sexually mature females to nests when combined with 3 KPZS. Nests baited with a mixture of DKPES and 3 KPZS (ratio 1∶29.8) attracted equal numbers of sexually mature females compared to an adjacent nest baited with 3 KPZS alone. When DKPES and 3 KPZS mixtures were applied at ratios of 2∶29.8 and 10∶29.8, the proportion of sexually mature females that entered baited nests increased to 73% and 70%, respectively. None of the sexually mature females released were attracted to nests baited with DKPES alone. These results indicated that DKPES is a component of the sex pheromone released by sexually mature male sea lamprey, and is the second biologically active compound identified from this pheromone. DKPES represents the first example that a minor component of a vertebrate pheromone can be combined with a major component to elicit critical sexual behaviors. DKPES holds considerable promise for increasing the effectiveness of pheromone-baited trapping as a means of sea lamprey control in the Laurentian Great Lakes.


Sleep apnea due to Kimura's disease of the larynx. Report of a case.
Kimura's disease is a benign chronic granulomatous disease which presents as a subcutaneous swelling in the head and neck area. The histopathological feature consists of granuloma-forming lymphoid follicles with eosinophil infiltration. Kimura's disease of the larynx is very rare, and only a few cases have been reported. We report a 14-year-old boy who presented with sleep apnea. Laryngological study revealed a submucosal swelling of the bilateral false vocal cord. Histopathological examination showed lymphoid hyperplasia with marked infiltration of eosinophils, which was diagnosed as Kimura's disease. After laser surgery, he had recurrent swelling of the false cord. The patient was given 30 mg of prednisolone, which was gradually tapered. The laryngeal swelling resolved, and the sleep apnea immediately improved after the treatment. However, a low dose of prednisolone was necessary to maintain the remission. Oral administration of pranlukast successfully supported the tapering of prednisolone.


[Evaluation of the efficacy and safety of combination of gemcitabine and nedaplatin for patients with HER-2 negative metastatic breast cancer].
Objective: To assess the therapeutic efficacy and safety of the gemcitabine combined with nedaplatin (GN) chemotherapy for metastatic human epidermal growth factor receptor-2 (HER-2) negative breast cancer patients. Methods: Forty-five patients with HER-2 negative recurrent metastatic breast cancer who had received prior adjuvant or neoadjuvant therapy with anthracycline and/or taxanes were enrolled. All the patients received GN regime from January 2014 to February 2019. The therapeutic efficacy was evaluated according to response evaluation criteria in solid tumors (RECIST) 1.1. The adverse response was evaluated and monitored according to common terminology criteria for adverse events (CTCAE). The progression-free survival (PFS) and overall survival (OS) and prognostic factors were also analyzed. Results: All of the 45 patients received 4 course GN, 1 of them achieved complete response, 21 achieved partial response. The objective response rate was 48.9 (95% CI: 33.7%-64.1%). Grade 3-4 hematological toxicities include leukopenia occurred in 10 (22.2%) of patients, neutropenia in 13 (28.9%) patients, and thrombocytopenia in 8 (17.6%) patients. The grade 3-4 hematological toxicities mainly manifested as nausea and vomiting, and the incidence was 4.4% (2/45). Among the 45 patients, 34 died, the median PFS was 5.1 (95% CI: 3.9-6.1) months and the median OS was 17.6 (95% CI: 13.1-20.9) months. Conclusion: The combination of gemcitabine and nedaplatin is an effective and tolerable treatment for metastatic breast cancer patients previously treated with anthracyclines and/or taxanes.


Effect of the calcium to phosphate ratio of tetracalcium phosphate on the properties of calcium phosphate bone cement.
Six different tetracalcium phosphate (TTCP) products were synthesized by solid state reaction at high temperature by varying the overall calcium to phosphate ratio of the synthesis mixture. The objective was to evaluate the effect of the calcium to phosphate ratio on a TTCP-dicalcium phosphate dihydrate (DCPD) cement. The resulting six TTCP-DCPD cement mixtures were characterized using X-ray diffraction analysis, scanning electron microscopy, and pH measurements. Setting times and compressive strength (CS) were also measured. Using the TTCP product with a Ca/P ratio of 2.0 resulted in low strength values (25.61 MPa) when distilled water was used as the setting liquid, even though conversion to hydroxyapatite was not prevented, as confirmed by X-ray diffraction. The suspected CaO presence in this TTCP may have affected the cohesiveness of the cement mixture but not the cement setting reaction, however no direct evidence of CaO presence was found. Lower Ca/P ratio products yielded cements with CS values ranging from 46.7 MPa for Ca/P ratio of 1.90 to 38.32 MPa for Ca/P ratio of 1.85. When a dilute sodium phosphate solution was used as the setting liquid, CS values were 15.3% lower than those obtained with water as the setting liquid. Setting times ranged from 18 to 22 min when water was the cement liquid and from 7 to 8 min when sodium phosphate solution was used, and the calcium to phosphate ratio did not have a marked effect on this property.


Lifelong mental health service use among 15-22 years old offenders: a document-based, mixed-methods descriptive study.
Among young offenders, psychiatric morbidity and comorbidity are common, but our knowledge about their use of mental health services during childhood and adolescence is scarce. We aimed to describe the lifelong use of mental health services of young offenders who have committed serious crimes. Using data on forensic psychiatric examinations of 42 Finnish offenders aged 15-22 years, we analysed the timing and typical patterns of their prior mental health service use with qualitative and quantitative content analysis and typification. Young offenders appeared in this study as children with plenty of perinatal and developmental risks, and risks related to their family situation and peer relations. Most subjects were described as having had emotional or behavioural symptoms, or both, since childhood. Involvement in mental health services was rare before the age of 7 years but increased markedly after that, staying on the same level during adolescence. Five categories of mental health service users were identified: (1) continuing service use around a decade (14.3%), (2) one brief fixed treatment (11.9%), (3) involuntary use of services (31.0%), (4) evasive use of services (21.4%) and (5) no mental health service use (21.4%). Young offenders had symptoms from early ages, but during childhood and adolescence, involvement in mental health services appeared for most as relatively short, repetitive or lacking. To help children at risk of criminal development, a multiprofessional approach, an early evidence-based intervention for behavioural symptoms and screening for learning problems, traumatic experiences and substance use are necessary. Results can help identify children and adolescents with a risk of criminal development, to develop mental health services and to plan further research.


Exome sequencing identified a novel de novo OPA1 mutation in a consanguineous family presenting with optic atrophy.
Inherited optic neuropathies are a heterogeneous group of disorders characterized by mild to severe visual loss, colour vision deficit, central or paracentral visual field defects and optic disc pallor. Optic atrophies can be classified into isolated or non-syndromic and syndromic forms. While multiple modes of inheritance have been reported, autosomal dominant optic atrophy and mitochondrial inherited Leber's hereditary optic neuropathy are the most common forms. Optic atrophy type 1, caused by mutations in the OPA1 gene is believed to be the most common hereditary optic neuropathy, and most patients inherit a mutation from an affected parent. In this study we used whole-exome sequencing to investigate the genetic aetiology in a patient affected with isolated optic atrophy. Since the proband was the only affected individual in his extended family, and was a product of consanguineous marriage, homozygosity mapping followed by whole-exome sequencing were pursued. Exome results identified a novel de novo OPA1 mutation in the proband. We conclude, that though de novo OPA1 mutations are uncommon, testing of common optic atrophy-associated genes such as mitochondrial mutations and OPA1 gene sequencing should be performed first in single individuals presenting with optic neuropathy, even when dominant inheritance is not apparent.


Role of Coatomer Protein I in Virus Replication.
Coatomer protein I (COPI) is well known as the protein coat surrounding vesicles involved in returning endoplasmic reticulum (ER)-resident proteins to the ER. COPI coats are also found in vesicles involved in other trafficking processes including endocytosis, autophagy and anterograde transport in the secretory pathway. In view of the diverse functions of COPI proteins, it is expected that they will affect virus replication, and many reports of such COPI involvement have now appeared. The experimental approaches most often employ specific siRNA to deplete COPI subunits or brefeldin A to block COPI activation. Here we briefly describe the results obtained with viruses in which COPI is found to have a role in replication. The results demonstrate that COPI affects viruses quite differently with effects observed in processes such as entry, RNA replication, and intracellular transport of viral proteins.


Randomised trial of pre-operative colour duplex marking in primary varicose vein surgery: outcome is not improved.
the ability of colour duplex to accurately locate incompetent venous sites has been widely published; its value in pre-operative marking in simple primary varicose vein surgery is evaluated in this study. prospective randomised controlled trial. regional vascular service, hospital patients. consecutive patients (149) undergoing primary varicose vein surgery where the only difference was one group of patients received duplex marking prior to surgery. Six weeks and 12 months post-operatively all patients had a colour duplex scan to determine the accuracy of surgery and the presence of residual/recurrent varicose veins and completed the Aberdeen, SF-36, and EuroQol quality of life questionnaires. varicose vein surgery. Duplex evidence of venous incompetence, quality of life measures using the SF-36 and Aberdeen Varicose Veins Questionnaire. pre-operative marking of primary varicose veins by skilled duplex ultrasonography does not improve the accuracy or recurrence rate following surgery. Quality of life improved significantly following surgery in both groups, however there was no difference in this improvement between the groups. the trial has not demonstrated any additional benefit of pre-operative colour duplex marking over that of clinical and hand held Doppler marking in terms of satisfactory varicose vein surgery performance at 6 weeks or 12 months in patients with primary varicose veins of the long saphenous system. It role in the short saphenous system is less clear.


Design, Synthesis, and Biological Evaluation of Coumarin-Triazole Hybrid Molecules as Potential Antitumor and Pancreatic Lipase Agents.
The design, synthesis, and investigation of antitumor and anti-lipase activities of some coumarin-triazole hybrid molecules are reported. The synthesis of these hybrid molecules was performed under microwave irradiation and conventional heating procedures. The newly synthesized hybrid molecules were investigated as inhibitors against four tumor cell lines (BT20 human breast carcinoma, SK-Mel 128 melanoma, DU-145 prostate carcinoma, and A549 lung carcinoma) and porcine pancreatic lipase (PPL). Most of these compounds showed notable antitumor activities against the tested tumor cell lines, and compounds 8i and 8l showed the best anti-lipase activity of 99.30 ± 0.56% and 99.85 ± 1.21%, respectively, at a concentration of 10 μM.


Chromosome 1 localization of the gene for CD34, a surface antigen of human stem cells.
CD34 is a surface antigen expressed on normal human hematopoietic stem cells, as well as on the blast cells of many patients with both lymphocytic and myelocytic leukemias. By Southern blot analysis of DNA from a panel of human x mouse somatic cell hybrids using a CD34 cDNA probe, we demonstrate that the gene for CD34 is located on human chromosome 1 in the 1q12----qter region.


Swelling-induced patterning in soft microchannels.
We study the effect of inflation on the swelling-induced wrinkling of thin elastic membranes in a set-up that is commonly used to create microchannels in lab-on-chip applications. Using a combination of experiments and associated numerical simulations, we demonstrate that the out-of-plane deformation of the inflated membrane and the resulting anisotropic stress lead to two distinct instabilities as the swelling progresses. The membrane first develops small-amplitude wrinkles that retain the cross-channel symmetry. Their wavelength depends on the pressure and is set in a process similar to the axisymmetric buckling of pressurised, uni-axially compressed cylindrical shells. As swelling increases, the membrane undergoes a secondary instability during which the wrinkles coarsen into large-amplitude folds whose morphology can be controlled by the degree of pre-inflation. We elucidate the fundamental mechanisms responsible for this behaviour and explain how inflation can be used as a control mechanism in the manufacture of microchannels.


Prevalence and risk factors of atherosclerotic renal artery stenosis in 1,200 chinese patients undergoing coronary angiography.
Atherosclerotic renal artery stenosis (ARAS) is a common and potentially reversible cause of end-stage renal failure. Our study aimed to determine the prevalence and predictors of ARAS in Chinese patients undergoing coronary angiography for suspected coronary heart disease (CAD), or for acute or chronic myocardial infarction. Selective renal arteriography was performed immediately after coronary angiography in 1,200 consecutive patients. Medical history and laboratory data were obtained before the procedure. Significant renal artery stenosis was defined as > or =50% narrowing of the luminal diameter. Uni- and multivariate logistic regression analyses were made to explore the association of the clinical and laboratory variables, including some items which had never or rarely been studied, with ARAS or CAD. Of the 1,200 patients, 840 were male and 360 female. Their mean age was 62 +/- 10 years. Low-grade (<50%) and significant coronary artery stenosis was found in 108 (9%) and 610 (51%) patients respectively. By multivariate logistic regression analysis, risk factors associated with the presence of coronary artery stenosis included male, older age, smoking, high serum concentration of low density lipoprotein, lipoprotein (a), and fast blood glucose > or =7.0 mmol/l. Significant ARAS was present in 116 (9.7%) patients, of which 20 (1.7%) were bilateral. The incidence of ARAS was similar in patients with suspected CAD or myocardial infarction. Multivariate logistic regression analysis showed the association of the clinical variables with ARAS included: older age, hypercholesterolemia, a more than 10-year history of hypertension, proteinuria and S(Cr) > or =133 micromol/l. The severity of ARAS is significantly related to the severity of coronary artery disease. ARAS is a frequent finding in Chinese patients undergoing coronary angiography, especially in patients with significant coronary artery stenosis and risk factors for ARAS. Renal arteriography can be a helpful examination in these patients.


Intraluminal implantation of rectal carcinoma successfully resected by endoscopy.
A 55-year-old Japanese woman presented at our hospital complaining of hematochezia 4 months after surgery for a rectal carcinoma. A proctoscopy revealed 2 protuberant lesions in the rectum, 5 mm anally from the anastomotic suture line. The diagnosis of adenocarcinoma was confirmed by biopsy. It was considered that these lesions were caused by intraluminal implantation from the primary rectal carcinoma. The patient underwent an endoscopic resection for these recurrent lesions and has remained stable, with neither recurrence nor metastasis, in the 7 years since the resection. For rectal carcinoma, we propose early follow-up by proctoscopy, namely within 4 months after surgery.


Facial nerve paralysis secondary to acute otitis media in infants and children.
Nowadays, facial paralysis is an uncommon complication of acute inflammation of the middle ear, with an estimated incidence of 0.005%; it was not so rare in the pre-antibiotic era, occurring in 0.5-0.7% of middle ear inflammatory processes. We would like to highlight this complication of acute otitis media, a common paediatric complaint. We present three new cases of facial palsy in children with acute otitis media and discuss the etiological mechanisms and different approaches to the treatment; a flow chart for facial paralysis in acute otitis media is also presented. Our three patients recovered completely after mastoidectomy (first two cases) and myringotomy with antibiotic therapy (third case). Facial paralysis is an uncommon complication of otitis media which requires appropriate care. Following our experience and revision of literature on the subject, antibiotic therapy and myringotomy are the first-line procedures. Surgery should be employed in case of acute or coalescent mastoiditis, suppurative complications and lack of clinical regression.


From philosopher to psychologist: the early career of Edwin Ray Guthrie, Jr.
Edwin R. Guthrie rose to prominence as a psychologist in the 1930s. His theoretical outlook was behavioristic. This approach came from his conviction that an objective method could be applied to a scientific treatment of mind. Prior to becoming a psychologist, he was a philosopher of mathematics. Guthrie was initiated into psychology by Stevenson Smith, from whom he learned a psychology of adjustment informed by comparative research, Columbia functionalism, and clinical psychology. Guthrie's first step into psychology was in collaboration with Smith on Chapters in General Psychology (S. Smith & E.R. Guthrie, 1921). To synthesize their own unique position on learning from the contemporary theory and research, they used the principle of association. This articles focuses on Guthrie's origin and his development into a learning theorist.


A phase I trial of gemcitabine and topotecan in previously treated ovarian or peritoneal cancer: a Gynecologic Oncology Group study.
To determine the maximum tolerated dose (MTD) of the combination of gemcitabine and topotecan in women with previously treated epithelial ovarian, peritoneal, or fallopian tube cancer. Patients with recurrent or persistent cancer after treatment with a platinum and paclitaxel-containing regimen were eligible for this study. Initial treatment was gemcitabine at a dose of 800 mg/m(2) on days 1, 8, and 15 and topotecan at a dose of 0.5 mg/m(2) on days 2-5, with cycles repeated every 28 days. Dose escalations were planned first for topotecan (Cohort I, Dose Levels 1-5) then for gemcitabine (Cohort II, Dose Levels 6-9) until the MTD was reached. Ten patients received a total of 29 cycles. When none of the first four patients could complete therapy as prescribed due to toxicity, doses for each drug were reduced by 1 day. The next six patients were treated at the modified schedule of gemcitabine days 1 and 8 and topotecan days 2-4 (Dose Level -1). Despite this modification, dose-limiting toxicities including neutropenia, thrombocytopenia, and stomatitis occurred at Dose Level -1, and the study was closed early. At both the initial dose schedule and an attenuated schedule, the combination of gemcitabine and topotecan produced dose-limiting toxicities in women with previously treated epithelial ovarian or peritoneal cancer.


Validation and Application of a Liquid Chromatography-Tandem Mass Spectrometry Method To Determine the Concentrations of Sofosbuvir Anabolites in Cells.
Sofosbuvir (SOF) is a highly efficacious and well-tolerated uridine nucleotide analog that inhibits the hepatitis C virus (HCV) NS5B polymerase enzyme. SOF is administered as a prodrug, which undergoes intracellular phosphorylation by host enzymes to a monophosphate, diphosphate, and finally a pharmacologically active triphosphate. In order to fully understand the clinical pharmacology of SOF, there is a great need to determine the intracellular phosphate concentrations of the drug. We describe the validation and utilization of a method to characterize SOF's disposition into various in vivo cell types, including hepatocytes, peripheral blood mononuclear cells (PBMC), and red blood cells (RBC). Standard bioanalytical validation criteria were applied to lysed cellular matrices, with a validated linear range of 50 to 50,000 fmol/sample for each phosphate moiety. The assay was utilized to collect the first data demonstrating concentrations of phosphorylated anabolites formed in PBMC, hepatocytes, and RBC in vivo during SOF therapy. Median concentrations in PBMC were 220 (range, 51.5 to 846), 70.2 (range, 25.8 to 275), and 859 (range, 54.5 to 6,756) fmol/10(6) cells in the monophosphate, diphosphate, and triphosphate fractions, respectively. In contrast, RBC triphosphate concentrations were much lower than those of PBMC, as the median concentration was 2.91 (range, 1.14 to 10.4) fmol/10(6) cells. The PBMC triphosphate half-life was estimated at 26 h using noncompartmental approaches, while nonlinear mixed-effect modeling was used to estimate a 69 h half-life for this moiety in RBC. The validated method and the data it generates provide novel insight into the cellular disposition of SOF and its phosphorylated anabolites in vivo.


Severe Hypoglycemia and the Use of Glucagon Rescue Agents: An Observational Survey in Adults With Type 1 Diabetes.
Severe hypoglycemia (SH) is the most frequent and potentially serious complication affecting individuals with type 1 diabetes and can have major clinical and psychosocial consequences. Glucagon is the only approved treatment for SH that can be administered by non-health care professionals (HCPs); however, reports on the experiences and emotions of people with type 1 diabetes associated with SH and glucagon rescue use are limited. This survey study demonstrated that an increasing number of individuals with type 1 diabetes have current and filled prescriptions for glucagon and have been educated about glucagon rescue use by an HCP. Despite this positive trend, challenges with SH remain, including a high level of health care resource utilization, considerable out-of-pocket expenses for glucagon kits, a high prevalence of hypoglycemia unawareness, and a negative emotional impact on individuals with diabetes. Nocturnal and exercise-related hypoglycemia were concerns for most survey participants.


The 100 most cited articles on oral cancer.
Citations analysis is one of the most widely used bibliometric tools to evaluate the academic importance of a study in a specific area of knowledge. The objective of this study was to identify the 100 most cited articles on oral cancer and to analyse their principal characteristics. We performed a literature search in the Web of Science database using the Science Citation Index Expanded tool to determine the number of citations of all articles on oral cancer identified up to 10 August 2017. The 100 most referenced articles were then selected, and the following information was gathered: ranking based on the number of citations; citation density; citations in Scopus; number and names of the authors; language and year of publication; country and institution of origin; financial support; journal of publication, with its impact factor, category and quartile; type of research; evidence level; and area of study. The number of citations of the 100 articles varied from 1959 to 165, and the number of authors varied from a single author to 23. The oldest article was from 1948, and the most recent was from 2013. All the studies were published in English, the majority (56%) was from the United States, and 80% were published in journals in the first quartile. The majority of articles were of studies that had received financial support, were published in journals with a high impact factor and were focussed on the aetiology and pathogenesis of oral cancer.


Sleep-Induced Glottis Closure in Multiple System Atrophy Evaluated by Four-Dimensional Computed Tomography.
Multiple system atrophy (MSA) is a progressive neurodegenerative disorder. Since patients with MSA often have sleep-related respiratory disorders including upper-airway obstruction and/or central sleep disturbance, appropriate evaluation of the upper airway especially during sleep may be indispensable. Fiberoptic laryngoscopy during diazepam-induced sleep has been reported for upper-airway obstruction verification. However, some patients cannot endure the uncomfortable sensation of the fiberscope. To address these issues, we devised a protocol of four four-dimensional computed tomography (4D-CT) for upper-airway evaluation during sleep. Here, we report the case of patient with MSA who was evaluated for upper-airway obstruction during sleep using 4D-CT. A 46-year-old man (height 1.60 m, weight 79 kg) was admitted to our neurological department for tracheal intubation because of a sudden onset of respiratory failure occurring at night. At the age of 45 years, he was diagnosed as MSA with predominant parkinsonism. As pulmonary disease had been excluded and his swallowing was normal, our differential diagnoses were central sleep apnea or obstructive sleep apnea related to his MSA or obstructive sleep apnea (SA) related to his obesity. A tracheostomy was done to maintain the airway after extubation. Polysomnography showed obstructive SA and not central SA. Awake fiberoptic laryngoscopy showed no upper airway obstruction but bilateral vocal abduction impairment (BVAI) during inspiration. To assess the spatial and temporal conditions of the upper respiratory tract-the patient could not tolerate sleep laryngoscopy-we carried out a 4D-CT. Reconstructed 4D-CT images of respiration during sleep showed clear abnormalities: glottis closure at the terminal stage of inspiration and subsequent velopharyngeal closure. As glottis closure does not occur normally in obesity patients, the cause of the respiratory failure in this patient was considered MSA-related sleep-induced airway obstruction. We decided to keep the tracheostoma, because BVAI in patients with MSA may be getting worse, although central apnea after tracheostomy may cause sudden central origin-related death; 4 months postoperatively, the patient had experienced no further airway-related complications. This report indicates that 4D-CT sequential upper-airway assessment during sleep is useful for determining the abnormalities causing obstructive SA in patients with MSA.


Characterization of the Maf family of polymorphic toxins in pathogenic Neisseria species.
In addition to harmless commensal species, Neisseria genus encompasses two pathogenic species, N. meningitidis (the meningococcus) and N. gonorrhoeae (the gonococcus), which are responsible for meningitis and genital tract infections, respectively. Since the publication of the first Neisseria genome in 2000, the presence of several genomic islands (GI) comprising maf genes has been intriguing. These GIs account for approximately 2% of the genome of the pathogenic Neisseria species and the function of the proteins encoded by maf genes remained unknown. We showed that maf genes encode a functional toxin-immunity system where MafB is a toxin neutralized by an immunity protein named MafI. A strain can harbor several MafB/MafI modules with distinct toxic activities. MafB toxins are polymorphic toxins with a conserved N-terminal region and a variable C-terminal region. MafB N-terminal regions consist of a signal peptide and a domain named DUF1020 that is only found in the genus Neisseria. MafB C-terminal regions are highly polymorphic and encode toxic activities. We evidenced the presence of MafB in the culture supernatant of meningococcal cells and we observed a competitive advantage for a strain overexpressing a MafB toxin. Therefore, we characterized a highly variable family of toxin-immunity modules found in multiple loci in pathogenic Neisseria species.


Vesicles of the intracellular and extracellular transport - key structures in the process of tissue differentiation towards bone and cartilage.
Differentiation of cells of the skeletal tissue, such as osteoblasts and chondrocytes, into mineralization-competent cells is a necessary step of the physiological process of bone and cartilage mineralization. Vascular cell calcification accompanies a pathological process of atherosclerotic plaque formation, which occurs due to trans-differentiation of vascular smooth muscle cells into cells resembling bone mineralization-competent cells. The activity of tissue-nonspecific alkaline phosphatase (TNAP), an enzyme necessary for physiological mineralization, is also induced in vascular cells in response to inflammation. TNAP acquires its mineralizing function when anchored to the plasma membrane (PM) of mineralizing cells and to the surface of vesicles derived from these cells. Numerous important reports indicate that various types of vesicles play a crucial role in initiating cell differentiation. In this review, we would like to highlight various functions of different types of vesicular structures of the cellular transport machinery such as intracellular vesicles (IVs), extracellular vesicles (EVs) or matrix vesicles (MVs) at distinct stages of both physiological and pathological processes of tissue differentiation.


Screening and diagnosis of inherited platelet disorders.
Inherited platelet disorders are important conditions that often manifest with bleeding. These disorders have heterogeneous underlying pathologies. Some are syndromic disorders with non-blood phenotypic features, and others are associated with an increased predisposition to developing myelodysplasia and leukemia. Platelet disorders can present with thrombocytopenia, defects in platelet function, or both. As the underlying pathogenesis of inherited thrombocytopenias and platelet function disorders are quite diverse, their evaluation requires a thorough clinical assessment and specialized diagnostic tests, that often challenge diagnostic laboratories. At present, many of the commonly encountered, non-syndromic platelet disorders do not have a defined molecular cause. Nonetheless, significant progress has been made over the past few decades to improve the diagnostic evaluation of inherited platelet disorders, from the assessment of the bleeding history to improved standardization of light transmission aggregometry, which remains a "gold standard" test of platelet function. Some platelet disorder test findings are highly predictive of a bleeding disorder and some show association to symptoms of prolonged bleeding, surgical bleeding, and wound healing problems. Multiple assays can be required to diagnose common and rare platelet disorders, each requiring control of preanalytical, analytical, and post-analytical variables. The laboratory investigations of platelet disorders include evaluations of platelet counts, size, and morphology by light microscopy; assessments for aggregation defects; tests for dense granule deficiency; analyses of granule constituents and their release; platelet protein analysis by immunofluorescent staining or flow cytometry; tests of platelet procoagulant function; evaluations of platelet ultrastructure; high-throughput sequencing and other molecular diagnostic tests. The focus of this article is to review current methods for the diagnostic assessment of platelet function, with a focus on contemporary, best diagnostic laboratory practices, and relationships between clinical and laboratory findings.


Epigenetic therapies in patients with solid tumors: Focus on monotherapy with deoxyribonucleic acid methyltransferase inhibitors and histone deacetylase inhibitors.
Epigenomics is the study of the gene expression changes due to epigenetic processes and not due to the deoxyribonucleic acid (DNA) base sequence alterations. The key mechanisms of epigenetic regulation include DNA methylation, histone modifications, and noncoding RNAs. Epigenetic alterations in cancer are predominantly linked with hypermethylation of promoters of the tumor suppressor genes, global DNA hypomethylation, and increased expression of histone deacetylases (HDAC). There is a growing need to investigate epigenetic patterns and to provide safe and effective, innovative therapeutic strategies for oncology patients, who did not improve on traditional anticancer regimens. The epi-drugs (e.g., DNA methyltransferase inhibitors, e.g., azacitidine and decitabine and HDAC inhibitors, e.g., vorinostat and romidepsin) have been approved for the clinical use. In this paper, we provide a brief overview of the mechanisms of action and targets for novel epi-drugs, focusing on their potential clinical applications in patients with solid tumors, resistant to standard oncology treatments.


Effects of graphene oxide and reduced graphene oxide nanomaterials on porcine endothelial progenitor cells.
Graphene oxide (GO) and reduced graphene oxide (rGO) have been widely used in the field of tissue regeneration and various biomedical applications. In order to use these nanomaterials in organisms, it is imperative to possess an understanding of their impact on different cell types. Due to the potential of these nanomaterials to enter the bloodstream, interact with the endothelium and accumulate within diverse tissues, it is highly relevant to probe them when in contact with the cellular components of the vascular system. Endothelial progenitor cells (EPCs), involved in blood vessel formation, have great potential for tissue engineering and offer great advantages to study the possible angiogenic effects of biomaterials. Vascular endothelial growth factor (VEGF) induces angiogenesis and regulates vascular permeability, mainly activating VEGFR2 on endothelial cells. The effects of GO and two types of reduced GO, obtained after vacuum-assisted thermal treatment for 15 min (rGO15) and 30 min (rGO30), on porcine endothelial progenitor cells (EPCs) functionality were assessed by analyzing the nanomaterial intracellular uptake, reactive oxygen species (ROS) production and VEGFR2 expression by EPCs. The results evidence that short annealing (15 and 30 minutes) at 200 °C of GO resulted in the mitigation of both the increased ROS production and decline in VEGFR2 expression of EPCs upon GO exposure. Interestingly, after 72 hours of exposure to rGO30, VEGFR2 was higher than in the control culture, suggesting an early angiogenic potential of rGO30. The present work reveals that discrete variations in the reduction of GO may significantly affect the response of porcine endothelial progenitor cells.


Altered gut microbiota-host bile acid metabolism in IBS-D patients with liver depression and spleen deficiency pattern.
Dysregulation of gut microbiota-host bile acid (BA) co-metabolism is a critical pathogenic factor of diarrhea-predominant irritable bowel syndrome (IBS-D). Traditional Chinese Medicine (TCM), instructed by pattern differentiation, is effective in treating IBS-D, in which liver depression and spleen deficiency (LDSD) is the most prevalent pattern. Still, it is unclear the linkage between the LDSD pattern and the BA metabolic phenotype. This study aimed to uncover the biological basis of the LDSD pattern from the BA metabolic perspective. Patients with IBS-D completed questionnaires regarding the irritable bowel severity scoring system (IBS-SSS), stool frequency, Stool Bristol scale, and Self-Rating Scales of mental health. Fasting blood and morning feces were collected to analyze the gut metagenome and BA-related indices/metabolites. IBS-D patients with LDSD had a higher incidence of BA overexcretion (41% vs. 23% non-LDSD) with significant elevations in fecal total BAs and serum BA precursor 7α-hydroxy-4-cholesten-3-one levels. Compared to controls or non-LDSD patients, LDSD patients had a featured fecal BA profile, with higher proportions of deoxycholic acid (DCA), 7-ketodeoxycholic acid, and lithocholic acid. It is consistent with the BA-metabolizing genomic changes in the LDSD gut microbiota characterized by overabundances of 7-dehydroxylating bacteria and BA-inducible genes (baiCD/E/H). The score of bowel symptoms (stool frequency and abdominal pain) showing greater severity in the LDSD pattern were positively correlated with bai-expressing bacterial abundances and fecal DCA levels separately. We clarified a differed BA metabolic phenotype in IBS patients with LDSD, which closely correlates with the severity of bowel symptoms. It demonstrates that gut microbiota and host co-metabolism of BAs would provide crucial insight into the biology of the LDSD pattern and its internal relationship with IBS progression.