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Acid and alkali as aids in differentiation of pathogenic Nocardia and for their isolation from clinical specimens.

The tolerance of Nocardia brasiliensis, Nocardia asteroides and Nocardia caviae to different concentrations of acids and alkalis was studied in this work. The purpose was to determine their use: (a) in the differentiation of species; (b) for decontamination of clinical materials during the isolation of pathogenic Nocardia. The results showed (1) that both 2 M NH3 and 0.05 M H2SO4 solutions are useful for differentiating species. They can also be used as a complementary method for identification. (2) That both 1 M NH3 and 0.05 M H2SO4 solutions were useful for decontamination of clinical materials during the isolation of pathogenic Nocardia.

Acetates↗

Voltage-dependent effect of a scorpion toxin on sodium current inactivation.

In voltage clamped nodes of Ranvier inactivation of the sodium permeability is slowed by toxin V from the scorpion Centruroides sculpturatus, by sea anemone toxin ATX II or by internally applied KIO3. The slow decay of the Na inward current is markedly accelerated if the test pulse is preceded by a depolarizing conditioning pulse followed by a 10-500 ms pause. This phenomenon was studied in detail, using conditioning pulses of varying amplitude and up to 15 s duration. In nodes treated with toxin V a 20 ms conditioning pulse to positive potentials was sufficient to produce a clear acceleration of the decay of the Na current and a reduction of the inward current remaining at the end of a 50 ms test pulse, i.e. a weakening of the toxin effect. In nodes treated with ATX II or internal KIO3 longer conditioning pulses were required. A similar effect of conditioning pulses on the decaying phase of the Na current was also observed in untreated fibres. To study the phenomenon quantitatively we fitted the decaying phase of the inward Na current with the equation INa = A exp(-t/tau 1) + B exp(-t/tau 2) + C. The effect of depolarizing conditioning pulses could be described as an increase of A, a decrease of B and C and a reduction of the time constants tau 1 and tau 2. I50/Ipeak, the normalised inward current remaining at the end of a 50 ms test pulse, decreased exponentially with increasing duration of the conditioning pulse to a steady-state value. The time constant tau and the steady-state value depended on the potential during the conditioning pulse. For nodes treated with toxin V, tau was 0.24 s at 0 mV and 12 degrees C and half inhibition occurred at -42 mV. The time constant tau was larger for nodes treated with ATX II or internal KIO3. At positive potentials, I50 was reduced to 20% of the control value in toxin V-treated nodes, but only to 70% in KIO3-treated nodes. Recovery from the effect of the conditioning pulse was studied by varying the pause between conditioning pulse and test pulse; recovery was 66-100% complete after 500 ms. The results are interpreted by assuming that a depolarizing conditioning pulse (a) accelerates inactivation of the sodium permeability and (b) causes dissociation of the toxin-receptor complex or transition into an inactive state. The latter effect occurs in toxin V-treated fibres but not in those treated with ATX II or KIO3.

Animals↗

The effect of diuretics on Na+-K+-ATPase and c-AMP levels in toad bladder epithelial cells.

Na+-K+-ATPase was inhibited by 1 times 10-4M ethacrynic acid and mercuderamide, and by 1 times 10-3M hydrochlorothiazide and furosemide. A modification of Gilman's (1970) protein displacement assay has been used to measure c-AMP levels in toad bladder epithelial cells. Vasopressin (50 mU/ml) caused c-AMP levels to rise from 4.27 to 9.27 pmol/mg protein. Ethacrynic acid had no effect on cellular c-AMP levels after 10 min exposure to the drug, but at 90 min caused a reduction of both basal and vasopressin stimulated levels. Furosemide caused an apparent rise in c-AMP levels, dilution ratio measurements indicated interference by this drug in the assay procedure, mecuderamide also caused substantial interference with the c-AMP assay. Hydrochlorothiazide had no effect on basal or hormone stimulated levels of c-AMP. It was concluded that the inhibition of sodium transport produced by ethacrynic acid in toad bladder is probably due to inhibition of adenylate cyclase, an effect not shared by other dieuretics.

Adenosine Triphosphatases↗

Harderian glands of golden hamsters: morphological and biochemical responses to thyroid hormones.

Manipulation of circulating levels of thyroid hormones modifies Harderian gland structure and porphyrin concentrations in male and female golden hamsters. Specifically, thyroxine (T4) and triiodothyronine (T3) induce the morphological conversion of the Harderian glands of females to approximate those of the male. Further, porphyrin concentrations are markedly decreased by this treatment. This effect occurs in ovariectomized animals as well, indicating that the gonads are not involved. Suppression of thyroid function by potassium perchlorate (KClO4) drastically reduces Harderian gland weight in both males and females. However, KClO4 decreases porphyrin levels in the Harderian glands of females and increases it in the male. Concurrently, KClO4 also induces a morphological conversion of the Harderian glands of males to the female type. This effect is evident in photoperiods of either 14:10 (h) or 8:16 (h).

Animals↗

Action of mercurials on the active and passive transport properties of sarcoplasmic reticulum.

The effect of Hg2+ and CH3-Hg+ on the passive and active transport properties of the Ca2+-Mg2+-ATPase-rich fraction of skeletal sarcoplasmic reticulum (SR) is reported. The agents abolish active transport, at 10(-5) and 10(-4) M concentrations, respectively. Addition of the mercurials was also shown to release actively accumulated Ca2+. The mercurials increase the passive Ca2+ and Mg2+ permeability in the absence of ATP at the same concentrations at which they inhibit transport. It is proposed that both effects are the result of direct binding of the mercurials to the SH groups of the Ca2+-Mg2+-ATPase pump, altering the conformational equilibria of the pump. The agents were also shown to increase the passive KCl permeability. The SR preparation consists of two vesicle populations with respect to K+ permeability, one with rapid KCl equilibration faciliated by a monovalent cation channel function and one with slow KCl equilibration. The mercurials increase the rates of KCl equilibration in both fractions, but produce higher rates in the fraction containing the channel function. The results are discussed in terms of pump and channel function and are compared with results for the electrical behavior of the CA2+-Mg2+-ATPase and other SR proteins in black lipid membranes, as presented by others.

Anilino Naphthalenesulfonates↗

Thyroid scintigraphy and perchlorate discharge test in the diagnosis of congenital hypothyroidism.

Quantitative thyroid scanning using low doses of technetium-99m sodium pertechnetate was performed on 147 infants (55 males and 92 females) with congenital hypothyroidism detected through the national neonatal screening programme. Thirty-two (21.8%) were athyrotic, while 62 (42.2%) had an ectopic thyroid and 53 (36%) had a eutopic gland with increased 99mTc uptake (mean 17%; range, 5%-38%). The perchlorate discharge test (PDT) was performed in nine of the infants with ectopic glands and 15 with eutopic glands; the findings were consistent with an organification defect in 22 cases (seven ectopic and 15 eutopic). Thyroid scintigraphy and PDT can add useful aetiological, genetic and prognostic information in the clinical evaluation of infants with congenital hypothyroidism detected by neonatal screening.

Choristoma↗

Inorganic-ion resistance by bacteria isolated from a Mexico City freeway.

Bacteria were isolated from soil samples, containing high exchangeable lead concentrations, obtained from a busy freeway in the México City metropolitan area. Forty-five selected strains (86.7% Gram-positive) had a single MIC distribution pattern for lead (800-1600 micrograms/ml lead nitrate) and were considered lead-resistant. The isolates showed variable levels of resistance to arsenate (86.7%), chromate (66.7%), cadmium (57.6%), and mercury (31.1%) ions. Multiple inorganic-ion resistance was shown by all strains.

Arsenates↗

Alterations in the prooxidant and antioxidant status of human leukemic T-lymphocyte MOLT4 cells treated with potassium chromate.

The involvement of reactive oxygen species in chromate-induced genotoxicity has been postulated. Because intracellular antioxidants help in eliminating the reactive species of oxygen, we have investigated both the prooxidant and antioxidant status of human leukemic T-lymphocyte MOLT4 cells exposed to nontoxic levels of chromium(VI) in culture. The cells treated with 0-->200 microM potassium chromate in a salts/glucose medium for 2 h were found to contain significantly lower levels of both small molecular weight and macromolecular antioxidants. In particular, the levels of glutathione and ascorbate were found to decrease with increased doses of chromate exposure in a dose-dependent manner. As little as 10 microM chromate was found to decrease these small molecular weight antioxidants significantly (p < 0.01). The macromolecular antioxidants, such as glutathione peroxidase, catalase, glutathione reductase, glucose-6-phosphate dehydrogenase and superoxide dismutase were also significantly (p < 0.01) decreased by exposing the cells to as little as 10 microM chromate. Concomitantly, there was a dose-dependent increase in intracellular H2O2 accumulation in cells exposed to chromium(VI). These results indicate that chromate-induced genotoxicity may be due, at least in part, to decreased levels of intracellular antioxidants in conjunction with an increased production of the reactive oxygen species.

Ascorbic Acid↗

Gamete development in Plasmodium berghei regulated by ionic exchange mechanisms.

Ionic regulation in the induction of exflagellation of Plasmodium berghei was investigated by culturing the parasites in various isotonic media. Of the salts tested, NaHCO3 exhibited the highest activity in inducing exflagellation, whereas KHCO3 showed no activity. In the absence of HCO3-, media containing monovalent cation (Na+, K+, Cs+, Rd+, choline+, lysine+, arginine+) and Cl- also induced exflagellation, but their activities were lower than that of NaHCO3. Anions of Br- or NO3- could be substituted with Cl-, whereas other anions such as I-, NO2-, SO4(2-), SCN-, H2PO4-, or HPO4(2-) failed to induce exflagellation, as did tetramethylammonium-Cl, CaCl2, MgSO4, MgCl2 and sucrose as well. These results suggest that the induction of exflagellation requires the presence of Na+ and HCO3- or monovalent, membrane-permeable cation and Cl- in the medium. Measurements of the efflux of H[14C]O3- or Cl- indicated that these anions were released from the cells into the NaCl or the NaHCO3 medium, respectively, probably by exchange in HCO3-/Cl-. Determination of intracellular ionic concentrations by electron microscopic X-ray microanalysis of cryopreserved specimens revealed that in the NaHCO3 medium, external Na+ (and probably HCO3-) enters the gametocytes by exchange with internal Cl- (and probably H+), whereas in Cl(-)-containing media, external unspecified cation and Cl- influx by exchange, probably with H+ and HCO3-. It is therefore suggested that two separate ion exchangers, i.e., Na(+)-dependent HCO3-(in)/Cl-(out) and nonspecific monovalent-cation-dependent Cl-(in)/HCO3-(out) exchangers, are involved in the induction of gametogenesis in P. berghei.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Vanadate inhibition of brain (Ca + Mg)-ATPase.

Vanadate was a potent inhibitor of the membrane-bound (Ca + Mg)-ATPase from rat brain, the concentration required for 50% inhibition under conditions optimal for enzymatic activity being 3 mu M. Vanadate inhibition increased with the MgCl2 concentration, half-maximal inhibition occurring at 2 mM MgCl2, near the MgCl2 concentration required for half-maximal activation of the ATPase activity. MnCl2 could substitute for MgCl2, and at concentrations of 1 mM (Ca + Mn)-ATPase activity was greater than (Ca + Mg)-ATPase activity, although sensitivity to vanadate was less. Vanadate inhibition increased also with the KCl concentration, half-maximal inhibition occurring at 8 mM, again near the concentration required for half-maximal activation of ATPase activity. By contrast, NaCl stimulated (Ca + Mg)-ATPase activity without potentiating vanadate inhibition. These effects of cations on ATPase activity and vanadate inhibition resemble properties of certain transport ATPases and thus suggest mechanistic and functional similarities.

Animals↗

Horse heart ferricytochrome c: conformation and heme configuration of high ionic strength acidic forms.

The absorption, circular dichroism, and resonance Raman spectra of horse heart ferricytochrome c in the presence of 0.2 M KCl, 0.1 M NaClO4, and 0.2 M KNO3, in the pH region 7 to 0.5, have been investigated to determine the nature and the course of the processes involved. As in the absence of salts (Myer, Y., and Saturno, A. F. (1990) J. Protein Chem, 9, 379-387), the change from neutral to low acidic pH's in the presence of salts is a three-step process: state IIIs----state IIIS,a----state IIS----state IS, with pKa's of 3.5 +/- 0.2, 2.2 +/- 0.2, and 1.1 +/- 0.2, and with two, one, and one number of protons, respectively. The addition of salts at neutral pH's has little or no effect on the protein conformation and the heme-iron configuration (i.e., they remain the same, low-spin hexacoordinated heme iron with a Met-80-Fe-His-18 axial coordination), but such addition does cause a slight tightening of the heme crevice and the enlargement of the porphyrin core. State IIIS,a is a folded state with about the same degree of folding and with a similar spin state and coordination configuration of iron, but the heme crevice is loosened and the porphyrin core is smaller. Both states IIS and IS are also essentially folded forms, but with a smaller degree of protein secondary structure. State IIS has a high-spin hexacoordinated heme iron with a water molecular and a protonated and/or hydrogen-bonded imidazole of his-18 as the two axial ligates; and state IS has a high-spin pentacoordinated heme iron, which is about 0.49 A out of the porphyrin plane, with a protonated and/or hydrogen-bonded imidazole nitrogen as the only axial ligate. The addition of anions causes the stabilization of the protein secondary structures and the state IIIa----state II transition. The mode of effectiveness of anions appears to be nonspecific (i.e., because of electrostatic shielding and/or disruption of salt bridges).

Animals↗

Identification of irradiated foods by the thermoluminescence of mineral contamination.

This investigation assessed the radiation-induced thermoluminescence (TL) of food minerals for identifying irradiated foods. Mineral contamination, rather than the inherent mineral content, of foods was studied. Positive identification of foods given 1-10-kGy doses depends on the contents and composition of the mineral contamination and the time span between irradiation and TL analysis. All 20 irradiated spice and herb samples could be identified without comparing them with unirradiated material. Three out of 60 lots of spices and herbs examined so far were so pure that the mineral contents were too low to allow TL analysis. If the soil where potatoes are grown contains feldspars, sprout inhibition treatment with 100-200-Gy doses can also be detected for up to one year, using the 90-250 degrees C low-temperature part of the TL spectrum for evaluation.

Aluminum Silicates↗

Activity of cathepsin D as affected by chemical and physical dry-curing parameters.

The effect of curing agents (nitrate, glucose, ascorbic acid and chloride) and physical parameters (temperature, water activity and pressure) on porcine muscle cathepsin D has been studied. Chloride (in the assayed range 0 to 75 g Cl-/L) showed a strong inhibitory effect. Nitrate (in the assayed range 0 to 800 mg/L) and high concentrations of ascorbic acid (4 to 8 g/L) slightly inhibited cathepsin D. However, its activity increased when glucose was added up to 4 g/L. Cathepsin D activity was maximal when incubated at 33 to 53 degrees C, was affected by a decrease in water activity and unaffected by pressure.

Animals↗

A possible role for cell proliferation in potassium bromate (KBrO3) carcinogenesis.

Accumulation of alpha 2u-globulin and induction of cell proliferation were examined in kidneys of rats exposed to KBrO3, KBr or NaBrO3 in their drinking water. Hyaline droplets observed after KBrO3 or NaBrO3 administration to male rats were specifically immunostained for alpha 2u-globulin. Increases in cell proliferation were found in the proximal tubules of male rats given KBrO3 or NaBrO3 but not KBr for 2, 4, and 8 weeks. No such change was evident in KBrO3-treated female rats or the distal tubules of any treated animal. The concordance between hyaline droplet accumulation and increased cell turnover suggests that KBrO3- and NaBrO3-induced cell replication in kidneys of male rats may result from alpha 2u-globulin nephropathy. Considering the fact that KBrO3 has genotoxic potential involving oxidative stress, we hypothesize that the induced cell proliferation might predominantly play an additive role in its carcinogenesis. Furthermore, the present data, showing similar effects of NaBrO3 on the rat kidney, are of direct significance to its risk assessment.

Alpha-Globulins↗

Detection of cellular receptors for Sendai virus in mouse tissue sections.

We have established a method for detecting virus receptors in the tissue sections and have been able to clarify the distribution of cellular receptors for Sendai virus in various mouse organs, including trachea, esophagus, large intestine and cerebrum. The virus receptors were detected in ciliated cells of the trachea, as to be expected for a pneumotropic virus. The lamina propria and connective tissue of the mouse esophagus and large intestine, immediately under the epithelium, contained an abundance at virus receptors, but very few were detected in the epithelium of these organs. In mouse cerebrum, the cells lining the ventricles had a large number of virus receptors.

Animals↗

Purification and hemagglutinating properties of egg drop syndrome 1976 virus.

We purified three populations of virus particles, F7, F9 and F17, with buoyant densities of 1.34, 1.33 and 1.29 g/ml, respectively, in CsCl equilibrium density gradients from cultures of chick embryo liver cells infected with the H-162 strain of the virus of egg drop syndrome 1976. F9 particles were infectious complete virions and most F17 particles were empty particles. F7 particles were less infectious, and had little capacity of hemagglutination (HA). HA titers were the same at 4 degrees and 37 degrees C and maximal between pH 6.4 and 8.4 and ionic strength from 0.14 to 0.54 M of NaCl. HA titer was inversely proportional to erythrocyte concentration. Potassium periodate destroyed markedly the infectivity of the virus and partially its HA activity at 37 degrees C. HA activity was stable at 56 degrees C or lower temperatures and destroyed at 80 degrees C. Trypsin, alpha-chymotrypsin, papain, ficin and neuraminidase had no effect on HA activity. Alpha-chymotrypsin destroyed the receptor for the virus on chicken erythrocytes, whereas trypsin and neuraminidase did not affect the receptor.

Adenoviridae↗