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Cryo-electron tomography (cryo-ET) enables visualization of protein complexes within their native cellular environment at molecular resolution. Most cells and all tissues, however, are too thick to be imaged directly by transmission electron microscopy (TEM). Overcoming this limitation requires the production of thin biological sections called lamellae. The procedure to obtain lamellae of cells, either seeded or grown directly on electron microscopy grids, requires cryo-focused ion beam (cryo-FIB) milling to thin the samples. This method faces an additional challenge when dealing with tissues and multicellular organisms, as these samples must be high-pressure frozen, which embeds the sample in a thick layer of ice. Nonetheless, lamellae can still be prepared from such samples by extracting a small volume and transferring it to a receiver grid for lamella preparation, a process called lift-out. Here, we describe the available workflows to produce lamellae by lift-out at cryogenic conditions and recent developments in gas injection system (GIS)-free approaches to the lift-out transfer. These advances expand the applications of cryo-ET, enabling the investigation of tissues and whole organisms in situ at molecular resolution.
The aim of the study was to investigate the effect of prebiotic fibres on appetite-regulating hormones, subjective feeling of appetite and energy intake in subjects with type 2 diabetes. Data presented are secondary outcomes of a study investigating the effect of prebiotics on glucagon-like peptide-1 and glycaemic regulation. We conducted a randomised and placebo-controlled crossover trial to evaluate the effects of 16 g/d of inulin-type fructans or a control supplement (maltodextrin) for 6 weeks in randomised order, with a 4-week washout period in-between, on appetite in thirty-five men and women with type 2 diabetes. Data were collected at visits before and after each treatment: plasma concentration of the satiety-related peptides ghrelin and peptide YY (PYY) were assessed during a standardised mixed meal. The subjective sensation of appetite was evaluated in response to an ad libitum lunch by rating the visual analogue scale. Twenty-nine individuals (twelve women) were included in the analyses. Compared to control treatment, the prebiotics did not affect ghrelin (P =0⋅71) or the ratings of hunger (P = 0⋅62), satiety (P = 0⋅56), fullness (P = 0⋅73) or prospective food consumption (P = 0⋅98). Energy intake also did not differ between the treatments. However, the response of PYY increased significantly after the control treatment with mean (sem) 11⋅1 (4⋅3) pg/ml when compared to the prebiotics −0⋅3 (4⋅3) pg/ml (P = 0⋅013). We observed no effect of inulin-type fructans on appetite hormones, subjective feeling of appetite or energy intake in patients with type 2 diabetes.
Mammary cancer is the second most common tumor worldwide. Small animal mammary neoplasms provide an outstanding model to study cancer in humans, as tumors in both share a similar environment, histopathologic features, and biological behavior. This study aims to investigate the percentage and microscopy of breast tumors in affected dogs and cats; its relationship to breed, age, and sex; and the immunohistochemical expression of estrogen receptor (ER), progesterone receptor (PR), Ki-67, and cytokeratin 8. Twenty-four females (12 dogs and 12 cats) and one male were examined from February 2018 to February 2020. The highest percentage of mammary neoplasia from the highest to the lowest manifested as tubular carcinoma, leiomyosarcoma, fibroadenoma, and cystic papillary carcinoma. The current study reported the second micropapillary invasive carcinoma in a male cat and the third lipid-rich carcinoma in a female cat. Although tubular carcinoma was the most common mammary neoplasm in cats, leiomyosarcoma was the most common in dogs. The immunohistochemical staining revealed diffuse and intense cytoplasmic immunoreactivity for cytokeratin 8 in lipid-rich carcinomas. However, moderate expression of ER in benign tumors and slight to moderate ER expression in malignant mammary lesions were reported. On the contrary, there was a negative PR expression in benign lesion. It could be concluded that a close relationship between ER expression and nuclear antigen Ki-67 was found.
Idiopathic basal ganglia calcification (IBGC) is a neuropathological finding known to manifest motor disturbance, cognitive impairment and psychiatric symptoms. Pathophysiology of psychiatric symptoms, however, remains controversial. Previous biochemical study suggests that dopaminergic impairment is involved in IBGC. We therefore performed positron emission tomography (PET) to elucidate the pre- and postsynaptic dopaminergic function and glucose metabolism in two IBGC patients.
Methods:
Case 1 is a 44 years old woman presented with disorganized thought, echolalia, verbigeration and parkinsonism. She was administered bitemporal electro-convulsive therapy (ECT). Case 2 is a 35 years old woman with persecutory delusion. Computed tomography showed bilateral symmetric calcification of striatum, globus pallidus and dentate nucei. Other causes of intracranial calcification were excluded. PET scans were obtained using [11C]-labeled 2β-carbomethoxy-3β-(4-flurophenyl)-tropane, [11C]-labeled raclopride and [18F] fluorodeoxyglucose.
Results:
The decreased binding potential was severe in bilateral head of caudate nuclei and anterior putamen. In case 1, the decline was also found in posterior putamen. There were widespread decreases of glucose uptake in frontal, temporal and parietal cortices bilaterally in case 1. Significant hypometabolism was observed in the right frontal, temporal and parietal cortices. After the ECT session, the previous areas of significant hypometabolism in the right hemisphere had improved. In case 2, there was no significant change of glucose metabolism in cerebral cortex.
Conclusions:
The difference in affected region within basal ganglia might be associated with the diverse clinical pictures in IBGC. Particularly, in the psychiatric manifestation, dopaminergic dysfunction in caudate nucleus and anterior putamen could be participated.
In this paper the Very High Energy (VHE) gamma-ray astronomy program at the University of Adelaide is described. VHE gamma rays with energies above ~5 × 1011eV are observed using the atmospheric Cerenkov technique. Results from the first three years observations at Woomera and the current upgrading of the telecope are described. The CANGAROO project, a collaboration between the University of Adelaide and a number of Japanese institutions, is also introduced.
The present study investigates the indications for transnasal endoscopic surgery in treating post-operative maxillary cysts.
Methods:
In this retrospective study, the records of 118 patients with post-operative maxillary cysts (88 unilateral and 30 bilateral) consisting of 148 procedures were reviewed.
Results:
A transnasal endoscopic approach was performed in 144 lesions (97.3 per cent). A combined endonasal endoscopic and canine fossa (external) approach was performed in 4 of 148 lesions, because the cysts were located distant from the nasal cavity and had a thick bony wall. A ventilation stent was placed in four patients (four cysts) to avoid post-operative meatal antrostomy stenosis. Recurrence was observed in five patients (4.2 per cent), all of whom subsequently underwent transnasal endoscopic revision surgery.
Conclusion:
Transnasal endoscopic surgery is an effective treatment for post-operative maxillary cyst with the exception of cysts located distant from the nasal cavity.
To validate a 204-item quantitative FFQ for measurement of nutrient intake in the Adventist Health Study-2 (AHS-2).
Design
Calibration study participants were randomly selected from the AHS-2 cohort by church, and then subject-within-church. Each participant provided two sets of three weighted 24 h dietary recalls and a 204-item FFQ. Race-specific correlation coefficients (r), corrected for attenuation from within-person variation in the recalls, were calculated for selected energy-adjusted macro- and micronutrients.
Setting
Adult members of the AHS-2 cohort geographically spread throughout the USA and Canada.
Subjects
Calibration study participants included 461 blacks of American and Caribbean origin and 550 whites.
Results
Calibration study subjects represented the total cohort very well with respect to demographic variables. Approximately 33 % were males. Whites were older, had higher education and lower BMI compared with blacks. Across fifty-one variables, average deattenuated energy-adjusted validity correlations were 0·60 in whites and 0·52 in blacks. Individual components of protein had validity ranging from 0·40 to 0·68 in blacks and from 0·63 to 0·85 in whites; for total fat and fatty acids, validity ranged from 0·43 to 0·75 in blacks and from 0·46 to 0·77 in whites. Of the eighteen micronutrients assessed, sixteen in blacks and sixteen in whites had deattenuated energy-adjusted correlations ≥0·4, averaging 0·60 and 0·53 in whites and blacks, respectively.
Conclusions
With few exceptions validity coefficients were moderate to high for macronutrients, fatty acids, vitamins, minerals and fibre. We expect to successfully use these data for measurement error correction in analyses of diet and disease risk.
When intrathecally or epidurally administered, α2-adrenoceptor agonists produce potent antinociception by affecting the activity of primary afferent fibres and spinal cord neurons. Recent reports have indicated that in dorsal root ganglion neurons, tetrodotoxin-resistant Na+ channels play important roles in the conduction of nociceptive sensation. We therefore investigated the effects of α2-adrenoceptor agonists on tetrodotoxin-resistant Na+ currents.
Methods
Using the whole-cell patch-clamp technique, we recorded tetrodotoxin-resistant Na+ currents from rat dorsal root ganglion neurons.
Results
Both clonidine and dexmedetomidine reduced the peak amplitude of the tetrodotoxin-resistant Na+ current concentration- and use-dependently. The concentration required for a half-maximal effect was significantly lower for dexmedetomidine (58.0 ± 10.2 μmol) than for clonidine (257.2 ± 30.9 μmol) at holding potential −70 mV. The current inhibitions induced by these agonists were not prevented by 1 μmol yohimbine, an α2-adrenoceptor antagonist. Both clonidine and dexmedetomidine shifted the inactivation curve for the tetrodotoxin-resistant Na+ current in the hyperpolarizing direction. The combinations clonidine with lidocaine and dexmedetomidine with lidocaine produced an additive blockade-type interaction on the tetrodotoxin-resistant Na+ current.
Conclusions
The results suggest that a direct inhibition of tetrodotoxin-resistant Na+ channels may contribute to the antinociceptive effects of clonidine and dexmedetomidine when used as additives to regional anaesthesia.
To examine a possible mechanism for the antinociceptive action of the N-methyl-d-aspartate receptor antagonist ifenprodil, we compared its effects with those of ketamine on tetrodotoxin-resistant Na+ channels in rat dorsal root ganglion neurons, which play an important role in the nociceptive pain pathway.
Methods
Experiments were performed on dorsal root ganglion neurons from Sprague–Dawley rats, recordings of whole-cell membrane currents being made using patch-clamp technique.
Results
Both drugs blocked tetrodotoxin-resistant Na+ currents dose dependently, their half-maximal inhibitory concentrations being 145 ± 12.1 μmol (ketamine) and 2.6 ± 0.95 μmol (ifenprodil). Ifenprodil shifted the inactivation curve for tetrodotoxin-resistant Na+ channels in the hyperpolarizing direction and shifted the activation curve in the depolarizing direction. Use-dependent blockade of tetrodotoxin-resistant Na+ channels was more marked with ifenprodil than with ketamine. When paired with lidocaine, these drugs produced similar additive inhibitions of tetrodotoxin-resistant Na+ channel activity.
Conclusions
The observed suppressive effects on tetrodotoxin-resistant Na+ channel activity may, at least in part, underlie the antinociceptive effects of these N-methyl-d-aspartate receptor antagonists.
Using very-high-frequency (VHF) plasma decomposition of SiH4 and pulsed gas technique, we have successfully prepared nanocrystalline silicon (nc-Si) quantum dots having average diameter of 8 nm and dispersion of 1 nm. The role of natural oxide is very important. It controls the size of nc-Si dots. Of particular interest is that the oxidation of these dots can be self limited, due to the stress induced near Si/oxide interface, which would allow further reduction of size and improvement in dispersion. This paper deals with the systematic study of oxidation process of nc-Si dots. Nc-Si dots formed in an Ar plasma with SiH4 gas pulses are deposited onto a Pt mesh The dots are then oxidized at 750, 800 and 850°C from 20 minutes to 15 hours. The dimensions of the residual nc-Si and the grown oxide are measured directly from the TEM micrographs and analyzed. For comparison, field oxide is investigated using ellipsometry. Retardation in the oxidation rate of nc-Si is observed. The mechanism of the reduction of oxidation rate in nc-Si is discussed taking into account the effect of stress.
A cold electron emitter has been made from nanocrystalline silicon (nc-Si) dots. Nc-Si dots are formed in the gas phase by very-high-frequency (VHF) plasma enhanced chemical vapor deposition (CVD). Electrons, accelerated by electric field, are ballistically transported through nc-Si and SiO2, then extracted into vacuum. Electron emission efficiency is optimized through varying nc-Si film thickness, surface roughness, and by short thermal oxidation.
A new method for the fabrication of nanocrystalline silicon (nc-Si) in SiH4 plasma with very-highfrequency (VHF; 144MHz) excitation is proposed to increase the deposition rate, to control the size, and to minimize size dispersion of nc-Si. Nanocrystalline silicon is formed in the gas phase of the SiH4 plasma cell by coalescence of radicals. Supplying Ar enhances the nucleation of nc-Si because of high efficiency of SiH4 excitation into SiH2 radicals resulting in the nucleation. The deposition rate is thus increased by a factor of 100 to 1012/cm2.h. At the low flow rate of SiH4, smaller nc-Si with small dispersion is obtained. Moreover, when pulsed-SiH4 is supplied into Ar plasma, the growth of nuclei is limited by the time when SiH4 flows. The size of nc-Si and its dispersion are adjusted by the duration of SiH4 gas pulse.
In this research we estimated the contribution of a major-gene
effect to the control of litter size in
hybrids between two local populations of the house musk shrew (Suncus murinus). Segregation
analysis was performed on the basis of a mixed polygene and major-gene
model. The model
presumes that two parental populations may differ from each other
in gene frequencies and in the
values of polygenic effects but not in the major-gene contribution of the
trait. Moreover, the
peculiarity of the trait – litter size – is taken into account.
This trait is not an individual attribute.
It characterizes the parental couple and may depend on the genotypes of
both parents. Results of
segregation analysis of a large hybrid pedigree of Suncus murinus
indicate that the parental
populations differ in the allele frequency of the major gene (one
population is homozygous, while
the other contains the two alleles in approximately equal proportions)
and
in the values of average
polygenic effects. Both major-gene and polygenic components are necessary
for the correct
description of litter size inheritance in interracial hybrids of
S. murinus, inasmuch as the exclusion
of either of them leads to a significant drop in likelihood. The
Elston–Stewart criterion also
confirms the Mendelian inheritance of the major gene.
A new method for the formation of nanocrystalline Si (nc-Si) in the SiH4 plasma using pulsed-H2 supply with very-high-frequency (VHF;144MHz) excitation is proposed to control the size of nc-Si. Nanocrystalline Si is formed in the gas phase of SiH4 plasma cell by coalescence of radicals. The principle of size control is based on the separation of nucleation and growth process. Supplying H2 into a SiH4 plasma enhances nucleation of nc-Si and suppresses growth rate of nc-Si. The nucleated nc-Si grows larger in a SiH4 plasma during the off state of the H2 supply. As the newly supplied H2 forces nc-Si grown in the previous cycle out of the plasma cell into the deposition chamber, the next nucleation of nc-Si is enhanced simultaneously. Using this method, we fabricated 8 nm-diameter nc-Si with small dispersion (±1 nm) successfully.
The Collaboration of Australia and Nippon for a GAmma Ray Observatory in the Outback operates two large telescopes at Woomera (South Australia), which detect the Čerenkov light images produced in the atmosphere by electronpositron cascades initiated by very high energy (~1 TeV or 1012 eV) gamma rays. These gamma rays arise from a different mechanism than at EGRET energies: inverse Compton (IC) emission from relativistic electrons.
The spoke-like images are recorded by a multi-pixel camera which facilitates the rejection of the large numbers of oblique and ragged cosmic ray images. A field of view ~3.5° is required. The Australian team operates a triple 4 m diameter mirror telescope, BIGRAT, with a 37 photomultiplier tube camera and energy threshold 600 GeV. The Japanese operate a single, highly accurate 3.8 m diameter f/1 telescope and high resolution 256 photomultipler tube camera. In 1998 a new 7 m telescope is planned for Woomera with a design threshold ~;200GeV.
We have proposed digital plasma processing for the fabrication of silicon quantum dots with grain size less than l0nm. By using the pulsed gas supply of SiH4 and H2 in the very-high frequency (VHF) plasma, we have clarified the role of atomic hydrogen in the nucleation, crystallization of nanocrystalline Si (nc-Si) as well as in the selective etching of amorphous Si to nc-Si. Recently, we have prepared nc-Si by employing an ultra-high-vacuum (UHV) chamber equipped with VHF plasma cells of SiH4 and H2. Flux rate of Si cluster depends significantly on the pressure of the plasma cell and VHF power. Spherical shaped nc-Si clusters less than 6nm in diameter have been observed by transmission electron microscopy (TEM). Infrared absorption measurements have clarified that the surface of nc-Si is covered by hydrogen. In an attempt to control the position of nuclei, we have prepared nc-Si on SiO2 with micro trenches, 40nm wide and 20nm deep, fabricated by electron beam exposure and electron cyclotron resonance (ECR) etching. It has been revealed by TEM observation that nc-Si are formed preferentially along micro trenches.
SI GaAs crystals submitted to single- or multi-step, ingot-or wafer-annealing are investigated using photoluminescence (PL) and photoacoustic spectroscopy (PA). The near-band-edge PL transitions are well resolved, with a neutral acceptor-bound exciton recombination displayed as a split doublet. The improvement induced by wafer-annealing is illustrated by the absence of additional defect-related transitions found after ingot-annealing. For the room temperature PA measurements, the intensity of a peak occuring at 1.39 eV is shown to lead to an estimation of the arsenic micro-defect density as evaluated by AB etching. The 1.39 eV PA band is also asserted to be the non-radiative recombination path of a 1.482 eV band found in the low-temperature PL spectra.
This paper reviews the results from two relatively new microwave heating studies under development at Ontario Hydro's Research Division. The first study involves the development of a hybrid kiln to process advanced ceramics at high temperatures. Using a combination of resistive heating elements and microwave heating, this new device has been operated to show significant process advantages in terms of time and energy savings. A description of the hybrid kiln is presented together with the test results for calcining and sintering a variety of ceramic materials.
The second investigation involves the use of microwave heating for the treatment of hazardous wastes. The selective heating ability of microwaves has been applied to reduce the environmental impact of wastes generated from industrial processes. An example is provided to illustrate the advantages and difficulties of applying this technology.
Implantation of Mg+ and C+ ions is carried out in bulk InP substrates using single or several energies up to 400 keV. The net carrier concentration profile at 300K is measured by capacitance-voltage (C-V) method. The ground and excited states binding energies of Mg and C acceptors are determined by low-temperature selective excitation of photoluminescence (PL). Additional sharp exciton-Iike emissions are detected after annealing of the samples. Their intensity is found to decrease with increasing Mg+ or C+ dose. Annealing-induced activation and/or formation of complex defect are the likely candidate mechanisms for the involved defect.