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The paper discusses possible application of the method of measuring the total content of organic carbon to evaluate the quality or reclaimed water. The paper gives data on the content of organic carbon, ratio of chemical consumption of oxygen and carbon in the atmospheric condensate, urine, condensate of urine low-temperature evaporation and portable water reclaimed from these sources.
Recently, the use of cryo-fixation followed by freeze-substitution has been shown to produce a better ultrastructural preservation of cellular components due to the rapidity of the fixation procedure and the lack of distortion during dehydration. When these techniques are applied to study the fine structure of axenically cultured trophozoites of E. histolytica, an improved preservation of cytoplasmic and nuclear components was found. The surface coat of the parasite appears much thicker and uniform than in conventionally treated samples. Also, the ground substance of the cytoplasm is packed with fibrogranular material, which is usually extracted by chemical fixation. The extremely fast fixation procedure allows the visualization of fusion and/or fission processes of cytoplasmic vacuoles and vesicles. Nuclear microtubules and cytoplasmic microfilaments are clearly identified. Low-temperature techniques allow not only a better preservation of the cell structure of E. histolytica parasites, but will also facilitate considerably future immunoelectronmicroscopical studies.
We investigated the preseasonal blooming conditions of the male flowers of Sugi (Cryptomeria japonica D. Don) which flowers commonly in spring. We also investigated atmospheric Sugi pollen during all seasons for three years from 1988 through 1991 to determine pollen scattering in autumn. Our results showed that a discernible amount of pollen is scattered from the middle of October into December; this scattering would be expected in the following spring. Almost none of the buds of the flowers collected in October and in November bloomed when incubated at temperatures ranging from 6 degrees C to 20 degrees C after low-temperature treatment to arise the blooms from dormancy. However, small numbers of the male flowers from various Sugi trees bloomed and scattered pollen under these conditions. We found that the ratio of the blooming male flowers was one-third or less per blossom. The ratio changed depending on the growth stage and incubation conditions. In conclusion, the density of airborne Sugi pollen scattering is not high in autumn in general. However, the density would be expected to be high enough to induce allergic reactions and clinical symptoms in some areas of Sugi growth, especially in a good harvest year.
A simple method of corneal cryopreservation, in which corneas were frozen at -18 to -24 degrees C, was examined. Rabbit and cat corneas were placed successively in solutions of 50% fetal calf serum in McCarey-Kaufman medium with an increasing glycerol and glucose content. They were then frozen and stored in a -20 degrees C domestic freezer. Rabbit corneas stored in this way were examined in vitro by light and scanning electron microscopy, and both rabbit and cat corneas were also assessed after orthotopic allotransplantation into adult recipient animals. Functional corneal grafts were obtained with rabbit and cat tissue that had been cryopreserved for 3-4 weeks and 1 week, respectively. Endpoint analysis (by light and scanning electron microscopy) of grafts that had survived for 50 days indicated the presence of an intact corneal endothelial monolayer. The corneal endothelium slowly degenerated as the storage time was increased. Importantly, however, the endothelium appeared to withstand the freezing and thawing processes and we conclude that it may be possible to store corneas at temperatures above -196 degrees C, without the need for complex, low-temperature cryopreservation systems.
The reaction cycle of bacteriorhodopsin in the purple membrane isolated from Halobacterium halobium has been studied by optical absorption spectroscopy using low-temperature and flash kinetic techniques. After absorption of light, bacteriohodopsin passes through at least five distinct intermediates. The temperature and pH dependence of the absorbance changes suggests that branch points and/or reversible steps exist in this cycle. Flash spectroscopy in the presence of a pH-indicating dye shows that the transient release of a proton accompanies the photoreaction cycle. The proton release occurs from the exterior and the uptake is on the cytoplasmic side of the membrane, as required by the function of bacteriorhodopsin as a light-driven proton pump. Proton translocating steps connecting release and uptake are indicated by deuterium isotope effects on the kinetics of the cycle. The rapid decay of a light-induced linear dichroism shows that a chromophore orientation change occurs during the reaction cycle.
Differential scanning calorimetry (DSC) and nuclear magnetic resonance (NMR) spectroscopy are applied to characterize the nonfreezable water molecules in fully hydrated D2O/sphingomyelin at temperatures below 0 degrees C. Upon cooling, DSC thermogram displays two thermal transitions peaked at -11 and -34 degrees C. The high-temperature exothermic transition corresponds to the freezing of the bulk D2O, and the low-temperature transition, which has not previously been reported, can be ascribed to the freezing of the phosphocholine headgroup in the lipid bilayer. The dynamics of nonfreezable water are also studied by 2H NMR T1 (spin-lattice relaxation time) and T2e (spin-spin relaxation time obtained by two pulse echo) measurements at 30.7 MHz and at temperatures down to -110 degrees C. The temperature dependence of the T1 relaxation time is characterized by a distinct minimum value of 2.1 +/- 0.1 ms at -30 degrees C. T2e is discontinuous at temperature around -70 degrees C, indicating another freezing-like event for the bound water at this temperature. Analysis of the relaxation data suggest that nonfreezable water undergoes both fast and slow motions at characteristic NMR time scales. The slow motions are affected when the lipid headgroup freezes.
The main objective for a reinvestigation of rhamnose was to devise a mechanistic link between the trapped electron detected previously and the secondary radicals observed at 77 K and at room temperature. Single crystals of rhamnose were X-irradiated at temperatures between 15 and 300 K and examined using ESR, ENDOR, and field-swept ENDOR techniques. After low-temperature irradiation a C3 H-abstraction radical is formed following the visible light-induced decay of the trapped electron. This species was previously assigned erroneously to a C2 H-abstraction species. At temperatures above 120 K, this radical deprotonates at the C3 hydroxy group. Furthermore, a C2 H-abstraction radical is formed following the thermally induced decay of the trapped electron. The C2 and C3 H-abstraction radicals did not convert into each other. A third radical species formed at low temperatures is a C5 H-abstraction radical. It is unstable above 250 K and decays without any apparent successor. The C2 and C3 H-abstraction radicals are formed thermally and photochemically from the parent trapped electron. The conversions are mediated by hydrogen atoms formed intermediately or by elimination of hydride ions. The thermal decomposition pathway requires further studies, in particular with respect to the possible role of water. Recently, Box et al. analyzed the site of the trapped electron in rhamnose crystals. The present results support the results obtained by these authors (Radiat. Res. 121, 262 (1990)). In particular, trapped electron vs proton distances closely match the conversion mechanisms suggested.
Ultrastructural immunoreactivities of alpha B-crystallin, glial fibrillary acidic protein (GFAP), ubiquitin, and vimentin in Rosenthal fibers (RFs) isolated from an Alexander's disease brain were investigated using nonosmium and low-temperature embedding technique. The morphology of RFs embedded in Lowicryl K4M resin was well preserved after treatment with 0.5% Triton X-100. alpha B-crystallin immunoreactivity was present in RFs of various sizes and was the strongest in loosely scattered deposits, which were considered to be the initial stage of RFs. Glial fibrillary acidic protein immunoreactivity in RFs was heavy, homogeneous throughout RFs, and equivalent to that in networks of glial filaments. Immunoreactivities of both alpha B-crystallin and GFAP were mainly restricted to the high electron-dense areas within RFs and were proved to exist close to each other by double immunolabeling. Rosenthal fibers were negative for vimentin. Ubiquitin immunoreactivity was relatively homogeneous in RFs with small diameters, but in RFs with large diameters, the immunoreactivity diminished in the center. Based on these observations, combined with the tendency of self-aggregation of alpha B-crystallin, it is conceivable that RFs are huge aggregation products of alpha B-crystallin involving GFAP, and that ubiquitination may be a consequent phenomenon, as it may be in other intracytoplasmic inclusions, such as neurofibrillary tangles and Lewy bodies.
The use of autologous blood in patients with organ transplantations is feasible only when the blood is stored in the frozen state, since the timing of the operation is not predictable. A 53 year-old patient with dilatative cardiomyopathy was selected for transplantation. By preoperative donation, four erythrocyte concentrates were prepared and frozen by high-glycerol low-temperature technique. The frozen blood was thawed, manually washed and made available for transfusion as required. The increment of hemoglobin after transfusion was 0.9 g/dl per unit. Quality control of the prepared erythrocyte concentrates was performed: Resistance to osmotic fragility of the erythrocytes was in the normal range, beginning hemolysis was found at 0.40 +/- 0.02% NaCl, complete hemolysis was found at 0.30 +/- 0.02% NaCl. The osmolarity of the supernatant fluid of the final washing was 0.314 +/- 0.116 mosmol/l. This case report stresses the feasibility to prepare and store autologous blood even if the time when the blood is needed cannot be determined.
Based on the literature data, the discussion covers the causes, preventive measures, and treatment of massive arterial air embolism, which is one of the most dangerous complications of artificial circulation (AC). Cerebral retroperfusion, hyperbaric oxygenation, low-temperature AC, and long-term administration of antihypoxic agents in the postoperative period have been considered most effective in the management of air embolism. The authors present their experience of successful management of four massive arterial air embolism cases, which is 0.22 percent of AC-assisted operations performed since 1984. Combined retrograde cerebral reperfusion and hypothermic AC have been used in the cases under discussion. Since it is impossible to determine the extent of embolism-associated brain damage and the limits of reliability of the applied therapy, it is essential in each embolism case to make use of all therapeutic means that are available in the given situation to combat this grave complication.
It has been shown that the resistance of muscular larvae of Trichinella spiralis to the effect of low temperatures depends on the conditions of freezing and composition of media possessing cryoprotective properties. It has been established that under optimal conditions (medium of microenvironment, conditions of freezing) during the storage in liquid nitrogen muscular larvae of T. spiralis preserve 50 to 60% of their resistance by the character of mobility and 11 to 15% by their capacity for reproduction. The question of creation of low-temperature bank for Trichinella is discussed.
We have identified a component of the eukaryotic RNA polymerase II transcriptional machinery that is more heat-labile than TFIID. DHFR transcriptional activity was severely reduced in 40 degrees C heat-treated extracts in which TFIID was fully active. This heat-labile activity was required for the transcription of both TATA box and non-TATA box promoters that are activated by the transcription factor Sp1. Gel mobility shifts indicated that Sp1 DNA binding activity was heat-labile, and the addition of purified Sp1 to 40 degrees C heat-treated extracts fully restored DHFR transcriptional activity. In contrast, the addition of Sp1 to 47 degrees C heat-treated extract did not result in transcriptional activity from the DHFR promoter. We conclude that reduction in Sp1 DNA binding activity is partially responsible for the heat-sensitive loss of DHFR transcriptional activity, but that a second essential activity is also inactivated by 47 degrees C heat-treatment. The discovery of this heat-labile component of Sp1 activation has two important implications in the analysis of transcriptional regulation. First, it demonstrates that heat-treated extracts are not appropriate for examination of the involvement of TFIID in the transcription of Sp1-activated promoters. Second, it explains the previously reported low-temperature optima for transcription from the DHFR promoter and demonstrates that transcriptional studies of Sp1-activated promoters should not be performed at 30 degrees C.
Using optical differential spectroscopy and EPR, a parallel study of light-induced electron transfer between the primary (X1) and secondary (X2) quinone-like acceptors in the preparations of reaction centers (RC) isolated from bacterial chromatophore membranes with sodium dodecyl sulfate was carried out. The data from direct measurements of the rate constant temperature dependence for the interaction between light-reduced X1 and X2 (KX1X2) are in good agreement with the data calculated from the kinetic analysis of dark reduction of photooxidized bacteriochlorophyll RC on the acceptors X1 and X2 (KX1X2 = 2.10(-1)S at 20 degrees; Ea = 11,8 kcal.mol-1 within the temperature range of 20 degrees-- -20 degrees). This evidence proves the efficiency of the previously used approach /1, 2/ for the evaluation of the X1-X2 interaction. The method proposed was used for a kinetic analysis of a low-temperature electron transfer from X1 to X2 in RC isolated with lauryldimethylaminoxide (KX1X2 = 2,3.10(2) S-1 at 20 degrees; Ea = 5,5 kcal.mol-1 within the temperature range of 10 degrees-- --70 degrees).
The IR-spectra (900-1800 cm-1) of films of native and denatured DNA were studied within the temperature range 4-100 degrees C at different relative humidity (r.h.). Two temperature intervals of S-like change of spectral parameters of absorption bands of sugar-phosphate chain of double-helical DNA are found on temperature relationships at high humidity. These two intervals point to two types of cooperative transitions of sugar-phosphate pattern. The band parameters of nitrous bases are changed only under high temperature transition. In the region of structural transitions of different subsystems of DNA hydrate hyperchromism was wound for the bands of sugar-phosphate chain and hydrate cover unlike hyperchromism specific of most nitrous bases bands. From the data obtained it is proposed to use the basic characteristics of temperature relationship of spectral parameters of the absorption bands of sugar-phosphate pattern and hydrate envelope in the region of low-temperature transition for the "in vivo" analysis of the DNA structure in complex biological systems.
The membrane-impermeable protein reagent p-diazonium benzene sulfonate (DABS) inhibits the mitochondrial respiratory chain in the cytochromes b-c1 region. It has been shown that DABS-treatment brings about a dramatic change in a low-temperature absorption spectrum of rat liver mictochondria, so that the long-wavelength alpha-band of ferrocytochrome b566 is sharply decreased, which is indicative of this hemoprotein modification. This observation suggest that cytochrome b566 is accessible to DABS at the outer surface of the coupling mitochondrial membrane.
Phospholipase A2 is shown to be activated by freezing-thawing, possibly due to changes in the state of lipid bilayer under the effect of both the temperatures themselves and physicochemical factors formed in the low-temperature range.
The structure of thin, vapor-deposited carbon films was characterized by transmission electron microscopy and electron diffraction. Selected area electron diffraction showed very weak and broad peaks, indicating that these carbons contain extremely small crystallites whose dimension in the crystallographic c-direction is about 8 to 10 a. The observed diffraction bands are (h, k, 1 = 0) type reflections, which suggests that individual crystallites consist of graphitic layer planes stacked in parallel groups but with no order between atoms in adjacent planes (turbostratic). The carbon films exhibit no preferred orientation, indicating that the small crystallites are randomly oriented in the film and that the films are therefore isotropic. The measured density (1.8 g/cm3) and the structure of the vapor-deposited carbons are accordingly similar to those of low-temperature isotropic (LTI) pyrolytic carbons.
Heat capacity and line width of proton magnetic resonance of collagen were studied by means of low-temperature calorimatry and high resolution NMR spectroscopy at different values of moisture in the temperature range of ice-water phase transition. Peculiarities of water phase transition in the presence of biopolymers were revealed. It was shown that a definite part of H2O in the "hydration layers" of the macromolecules (0.35 g of H2O/g of protein) is not transformed into ice even at a temperature decrease up to 4 K. The NMR data show that the total mobility of the bound water molecules remains high enough up to the water concentration at which the inner hydration layer of the macromolecule is formed. After the formation of hydrogen-bound water chains in the hydrate layers of collagen the mobility of the bound water molecules decreases.