Amebic keratitis in a wearer of disposable contact lenses due to a mixed Vahlkampfia and Hartmannella infection

PURPOSE: To support the hypothesis that Acanthamoeba is not a unique cause of amebic keratitis, we report a case of amebic keratitis in which viable Acanthamoeba could not be isolated from corneal tissue. Vahlkampfia and Hartmannella, two other genera of free-living ameba, were isolated, however, using prolonged culture. METHODS: A 24-year-old wearer of soft contact lenses had keratitis. Extensive histologic and microbiologic investigations were performed on corneal scrape, biopsy, and keratoplasty tissue. Contact lenses, storage case, and the home water supply, where contact lens hygiene was practiced, were examined for the presence of micro-organisms. RESULTS: No viruses, pathogenic bacteria, or fungi were detected from corneal tissue samples. Amebae were observed using light and electron microscopy, but these could not be unequivocally classified using immunocytochemical staining. Viable Vahlkampfia and Hartmannella, but no Acanthamoeba, were isolated from the corneal biopsy sample. Indirect immunofluorescence with a range of polyclonal rabbit antisera raised against axenically cultivated stains of the three amebal genera was unhelpful because of cross-reactivity. A diverse range of micro-organisms was present within the storage case, including the three amebal species. Amebic cysts also were associated with the contact lens. CONCLUSION: A mixed non-Acanthamoeba amebic keratitis has been identified in a wearer of soft contact lenses where lack of storage case hygiene provided the opportunity for the free-living protozoa Vahlkampfia and Hartmannella to be introduced to the ocular surface. When Acanthamoeba-like keratitis occurs, but where Acanthamoeba cannot be isolated using conventional laboratory culture methods, alternate means should be used to identify other amebae that may be present. Polyclonal immunofluorescent antibody staining was unreliable for generic identification of pathogenic free-living amebae in corneal tissue.     

Chemiluminescent detection of mRNAs on northern blots with digoxigenin end-labeled oligonucleotides

To establish a simplified, nonradioactive approach for identifying mRNAs on Northern blots, antisense oligonucleotides have been used as probes in combination with chemiluminescence-based detection. Oligonucleotides (approximately 32-mer) were end-labeled with digoxigenin (DIG) and used in conjunction with adamantyl 1,2-dioxetane aryl phosphate substrates (Lumigen PPD and CSPD). Oligonucleotides were designed as probes for several mRNAs in tissues of rats and mice, including the mitochondrial uncoupling protein, lipoprotein lipase, GLUT1, GLUT4, and beta-actin. Uncoupling protein mRNA was detected in total RNA from brown adipose tissue with a 32-mer DIG-labeled oligonucleotide, within 2 min of exposure to film. This mRNA could also be detected when as little as 250 ng of total RNA was applied to the gel, following 4 h exposure to film, and was present only in brown fat. The mRNA for lipoprotein lipase was detectable with a 30-mer DIG-labeled oligonucleotide in 1 micrograms of total RNA from mouse heart, within 2 h of exposure. The mRNA for the GLUT1 glucose transporter was detected in total RNA from rat midbrain using a 32-mer DIG-labeled oligonucleotide, while beta-actin mRNA was detected with a 30-mer oligonucleotide. The mRNA for the insulin-sensitive glucose transporter GLUT4 was detected with a 32-mer DIG-labeled oligonucleotide and found only in those tissues in which glucose uptake is stimulated by insulin. The speed of detection was greater with CSPD and was augmented by exposure of membranes to film at 37 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative investigation of antibacterial yet biocompatible Ag-doped multicomponent coatings obtained by pulsed electrospark deposition and its combination with ion implantation

The aim of this work is to obtain antibacterial yet biocompatible coatings using pulsed electrospark deposition (PED). For this purpose new composite electrodes were fabricated from reaction mixtures Ti–C–20%Fe-10%Ca₃(PO₄)₂–3.4%Mg–X%Ag with different amount of antibacterial component (X = 0, 0.5, 1.0, 1.5 and 2.0 at% of Ag) using self-propagating high-temperature synthesis method. The electrodes consisted of TiC grains surrounded by TiFe₂ and TiFeP intermetallic matrix, CaO and MgO inclusions, and Ag-based phase. The influence of Ag content on the electrode mass transfer kinetics was studied by comparing the total substrate weight gain and electrode mass loss during PED. The structure, elemental composition, and surface roughness of coatings were studied by means of X-ray diffraction, scanning electron microscopy, and optical profilometry. The coatings were characterized in terms of Ag⁺ ion release, mechanical and electrochemical properties, as well as biocompatibility. The antibacterial characteristics of Ag-doped PED coatings were compared with those obtained by PED using Ag-free electrode and then implanted with Ag⁺ ions. The results indicated that an increase in the Ag content in electrode leads to a decrease in electrode erosion and substrate weight gain, but the efficiency of the PED process increases. Doping with a small amount of Ag (≤ 1 at%) resulted in 100% antibacterial effect against both gram-positive S. aureus and gram-negative E. сoli bacteria. In addition, the dynamics of МС3Т3-Е1 cell proliferation on the surface of PED coatings with 0.6–0.7 at% of Ag was similar to that in control samples, hereby indicating their biocompatibility. The coating biological characteristics were discussed based on the results of Ag⁺ ion release and electrochemical tests.     

The genetic effects in the somatic cells of persons working under chronic irradiation of different intensities after the accident at the Chernobyl Atomic Electric Power Station

A complex genetic study of two groups from of Chernobyl NPP personnel (from "Shelter" unit and 3rd Block) has been carried out using classical cytogenetic and GPA methods. The first group was the most vulnerable from the viewpoint of accumulated dose (exceeding 25 cGy for the moment of study). Positive correlation between individual and group frequencies of cytogenetic markers of irradiation (stable and unstable chromosomes aberrations) and NO mutations in the GPA locus was found.     

Boosted Responsivity and Tunable Spectral Response in B-Site Substituted 2D Ca2Nb3−xTaxO10 Perovskite Photodetectors

2D Dion-Jacobson perovskite oxides, featuring fascinating optical and electric properties, exhibit great potential in optoelectronic devices. However, the device sensitivity and spectral selectivity are limited. Herein, B-site substituted calcium niobate Ca₂Nb_3−xTa_xO₁₀ (x = 0, 0.5, 1, 1.5) nanosheets are prepared by liquid exfoliation. The photodetectors (PDs) based on these nanosheets exhibit tunable spectral response by tailoring the band gap of the nanosheets. All the Ta-substituted PDs show increased photocurrent and enhanced responsivity, among which the Ca₂Nb_2.5Ta_0.5O₁₀ PD exhibits the optimal performance with a photocurrent of 31.4 µA, a high on–off ratio of 5.6 × 10⁴ and a boosted responsivity of 469.5 A W⁻¹ at 1.0 V toward 295 nm, which is over 7000-fold higher than that of pristine Ca₂Nb₃O₁₀ PD. It is proposed that the significantly optimized responsivity is ascribed to the enhanced photoconductive gain that mainly originates from the introduction of the trap states by the B-site substitution. Nevertheless, excess substitution is detrimental to the responsivity and the response speed. This work demonstrates that the rational control of B-site substitution tailors the band gap and modulates the charge-carrier behaviors in 2D perovskite oxides, which provides an effective avenue for achieving high-performance PDs with tunable spectral response and excellent responsivity.     

Biochemistry and hematology at decompression sickness: a case report

A 24-year-old hospital corpsman, a volunteer in a series of dry chamber air dives to a simulated pressure equivalent to 188 FSWG (57.3 MSWG), developed left knee pain shortly after standard decompression. A tentative diagnosis of decompression sickness was made and recompression therapy was initiated with alleviation of pain occurring at 60 FSWG (18.3 MSWG). A U.S. Navy Treatment Table "5 (oxygen breathing) regimen was then selected and completed uneventfully. The subject had been undergoing biomedical evaluation for several days prior to diving; thus, a clinically diagnosed case of dysbarism with subsequent treatment was available for study. This individual was then monitored for a 10-d period. The acute phase of decompression sickness was characterized by a marked shortening of clotting time and a thrombocytopenia with accompanying increased platelet aggregates. The recovery phase was categorized by a variety of hematological and bio-chemical changes. Hemodilution, an elevated megathrombocyte index, and a tendency toward eosinopenia were evident for most of the 10-d observation period. Other persistent alterations detected during this period included a relative hyperglycemia, depressed urine Na+/K+, and increased ketosteroid excretion. These observations indicate that abatement of pain after treatment of dysbarism can be followed by the onset of a variety of biochemical and hematological changes. Moreover, complete recovery may require upwards of 10 d.     

Multifunctional nanostructured coatings: Formation,structure, and the uniformity of measuring their mechanical and tribological properties

The state of the art of the studies in the field of the development and certification of novel multifunctional nanostructured coatings having a wide spectrum of applications is reviewed. The main tendencies in the optimization of the compositions and properties of the coatings are described, and the modern methods of diagnostics of the nanostructured coatings are considered.     

Effect of synthesis conditions and surrounding medium on luminescence properties of YVO4:Eu3+nanopowders

Nanocrystalline yttrium vanadate doped with europium ions powders were synthesized via sol-gel method based on decomposition of metal-polymer complex. X-ray diffraction analysis showed that samples had pure tetragonal phase without any impurities. Scanning electron microscopy and static light scattering technique were used to study morphology and size of prepared nanoparticles. Average diameter of the nanoparticles was about 40 nm. The changes in structural and luminescence properties were observed as a function of the first and second calcination temperature. The optimal conditions for synthesis of nanoparticles were determined as Т1=500 °С, t1=1 h; Т2=950 °С, t2=1.5 h. The effect of different media surrounding the nanoparticles on their luminescence properties and lifetime was investigated and discussed in terms of effective refractive index. It was found that the observed lifetime of YVO4:Eu3+ 5 at.% nanophosphor was decreased from 0.64 ms in air(nmed=1) to 0.45 ms in chalcogenide glass As39S61(nmed=2.39).     

Rapid, activation-induced redistribution of ionotropic glutamate receptors in cultured hippocampal neurons

We have examined the membrane localization of an AMPA receptor subunit (GluR1) and an NMDA receptor subunit (NR1) endogenously expressed in primary cultures of rat hippocampal neurons. In unstimulated cultures, both GluR1 and NR1 subunits were concentrated in SV2-positive synaptic clusters associated with dendritic shafts and spines. Within 5 min after the addition of 100 microM glutamate to the culture medium, a rapid and selective redistribution of GluR1 subunits away from a subset of synaptic sites was observed. This redistribution of GluR1 subunits was also induced by AMPA, did not require NMDA receptor activation, did not result from ligand-induced neurotoxicity, and was reversible after the removal of agonist. The activation-induced redistribution of GluR1 subunits was associated with a pronounced (approximately 50%) decrease in the frequency of miniature EPSCs, consistent with a role of GluR1 subunit redistribution in mediating rapid regulation of synaptic efficacy. We conclude that ionotropic glutamate receptors are regulated in native neurons by rapid, subtype-specific membrane trafficking, which may modulate synaptic transmission in response to physiological or pathophysiological activation.