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Beneficial effects of GM1 ganglioside on photochemically-induced microvascular injury in cerebral cortex and hypophysis in rat.

Gangliosides, the glycophospholipids which are abundantly present in the central nervous system, have been shown to stimulate neuronal regeneration and counteract the deleterious effects of ischemia on cerebral neurons. The further elucidate the mechanism of action of gangliosides in cerebral ischemia, we investigated the influence of GM1 ganglioside in the model of photochemically-induced microvascular injury in rat brain. The animals were injected with rose Bengal and illuminated through cranium with halogen lamp. This treatment resulted in the development of microthrombi and alterations in endothelial cells in the microvessels. Administration of 20 mg/kg GM1 ganglioside, 1 h before the photochemical reaction, largely reduced subsequent microvascular damage. In conclusion, the GM1 ganglioside is able to prevent microvascular damage in the central nervous system.

Animals↗

Vitamin A status of young children in Southern Brazil.

A comprehensive survey was carried out to assess the vitamin A status of preschool children of poor migrant families in the periurban population of Ribeirao Preto, a typical agricultural town in the sugarcane and coffee region of the State of Sao Paulo in Southern Brazil. The intake of vitamin A and carotenoids from the rice and bean based diet of these children is considered low and appears to influence blood concentrations and liver reserves of this vitamin. With respect to plasma vitamin A, 1.8% of the children had a deficient level (less than 10 micrograms%), whereas 48.8% of the children had a low level (less than 20 micrograms%). Most children with inadequate plasma vitamin A (less than 20 micrograms%) responded positively to a massive dose of 200,000 IU vitamin A, suggesting that these children may be at risk of having low liver stores of vitamin A. Rose Bengal staining test and rapid dark adaptation time did not indicate definite signs of conjunctival xerosis and night blindness among these children. No ocular evidence of hypovitaminosis A was found in the children studied, but marginal or inadequate vitamin A status appears to be a common public health problem among young children in this region of Brazil.

Anthropometry↗

Photochemically induced spinal cord injury in the rat.

We have developed in the rat a minimally invasive model of reproducible spinal cord injury initiated photochemically. With the exposed spinal column intact, 560 nm irradiation of the translucent dorsal surface induces excitation of the systemically injected dye, rose Bengal, in the spinal cord microvasculature. The resultant photochemical reaction leads to vascular stasis. Histopathological changes at 7 days include hemorrhagic necrosis of the central gray matter, edematous pale-staining white matter tracts and vascular congestion. At the level of cord irradiation (T8) the entire cord thickness is necrosed except for the periphery of the anterior funiculus. Voluntary motor function is consistently lost in the subacute phase of injury.

Animals↗

On the mechanism of action of choline acetyltransferase.

The substrate specificity of choline acetyltransferase (EC 2.3.1.6) isolated from squid ganglia was investigated. The enzyme catalyzed the acetylation of choline and aminocholine but not of homocholine. In D(2)O solution there was considerable slowing of the transacetylation reaction. Photo-oxidation in the presence of methylene blue or rose bengal rapidly inactivated the enzyme, suggesting involvement of a histidine residue in the catalytic site. It seems likely that general-base catalysis by imidazole enhances the ability of enzyme-bound choline (or ammoniumcholine) to react with a thiolester group. Attempts to isolate an acetylthio-enzyme intermediate after incubation with [(14)C]acetylcoenzyme A were unsuccessful. A possible mechanism for the action of choline acetyltransferase is proposed.

Acetyl Coenzyme A↗

Photosensitized degradation of losartan potassium in an extemporaneous suspension formulation.

During development of an extemporaneous suspension formulation for losartan potassium, previously unknown degradation products were observed in experimental suspensions prepared in a commercial cherry syrup vehicle. These degradates increased rapidly when analytical solutions prepared from that suspension were exposed to ambient light. The structures of the degradates were determined using a combination of preparative HPLC, LC/MS, (13)C and (1)H NMR (1D and 2D), and mechanistic chemistry. Each degradate results from destruction of the imidazole ring of losartan. Formation of the two major degradates required exposure to light (UV or visible) and the presence of oxygen. Experiments using Rose Bengal (a singlet oxygen photosensitizer) and 1,4-diazabicyclooctane (DABCO; a singlet oxygen quencher) established that the major photodegradates are formed via the intermediacy of singlet oxygen. The identity of the photosensitizer in the formulation was not unequivocally determined; however, the experiments implicated the artificial flavoring in fulfilling this role.

Angiotensin II Type 1 Receptor Blockers↗

Differentiation of Brucella abortus and Yersinia enterocolitica serotype 09 infections in cattle: the use of specific lymphocyte transformation and brucellin skin tests.

The lymphocyte transformation test (using an in vitro whole-blood lymphocyte stimulation procedure) and the Brucellin skin test were applied to five heifers infected with virulent Brucella abortus strain 544, five cows inoculated with Yersinia enterocolitica serotype 09, and four non-exposed cows. Lymphocytes from Brucella-inoculated animals persistently gave very high blastogenic reactions indicative of active Brucella infection. The test was persistently negative in Yersinia-infected and non-exposed cattle. Four of the five cows infected with Yersinia enterocolitica type 09 and all four control cattle were persistently negative to the delayed hypersensitivity skin reaction with brucellin. All cattle infected with Yersinia enterocolitica type 09 were strongly positive to the Rose Bengal, Serum agglutination, Complement fixation and Antibovine globulin tests using Brucella abortus antigens. One lactating cow infected with Yersinia enterocolitica type 09 was positive to Brucella milk ring test. These results indicate that standard Brucella serological tests are unreliable in differentiating the two infections in cattle and that both the Lymphocyte transformation and brucellin skin tests could be used to differentiate bovine brucellosis from yersiniosis.

Agglutination Tests↗

Singlet oxygen: a primary effector in the ultraviolet A/near-visible light induction of the human heme oxygenase gene.

Both singlet oxygen and the hydroxyl radical are generated in mammalian cells by UVA (320-380 nm) and possibly near-visible (380-420 nm) radiation. We have modulated the cellular levels of these two reactive oxygen species in order to compare their involvement in the induction of the human heme oxygenase (HO) gene by broad spectrum UVA/near-visible light (UVA/NVL). Irradiation in deuterium oxide (in which singlet oxygen has a longer half-life) enhances the broad spectrum UVA/NVL induction of this gene. Sodium azide and L-histidine which are scavengers of both singlet oxygen and the hydroxyl radical reduce the fluence-dependent accumulation of HO mRNA, while compounds which are only hydroxyl radical scavengers, namely, mannitol and dimethyl sulfoxide do not. Rose Bengal, a known generator of singlet oxygen, also increases the HO mRNA levels, and this induction is enhanced in deuterium oxide. We conclude that the observed effects of deuterium oxide and singlet oxygen scavengers on HO mRNA levels are not due to a nonspecific effect on transcription but that singlet oxygen is a primary effector in the UVA/NVL induction of the human heme oxygenase gene.

Azides↗

Brucellosis: the first case of King Chulalongkorn Memorial Hospital and review of the literature.

Brucellosis remains a major zoonotic disease worldwide. It has never been reported at King Chulalongkorn Memorial Hospital (KCMH). The authors describe the first case of brucellosis in KCMH, and also review all previous reports in Thailand. The presented case was a 52-year-old Thai man, living in Phetchabun Province, who was diagnosed with idiopathic pulmonary fibrosis two years prior to admission. He presented with prolonged fever, dry cough, weight loss of eight kg over three months, hepatosplenomegaly, and pancytopenia. Blood and bone marrow cultures grew Brucella melitensis at 72 hours of incubation. A slide agglutination (Rose Bengal) test was also positive for Brucella antibody. He had been exposed to contaminated placenta of his goats that had spontaneous abortion in the past few months before his illness. The patient was successfully treated with gentamicin, doxycycline, and rifampicin. Clinicians should have a high index of suspicion when evaluating patients presenting with prolonged fever and having an exposure risk of brucellosis.

Animals↗

[Formation of 8-hydroxydeoxyguanosine from 2'-deoxyguanosine and its decomposition by active oxygen].

8-hydroxydeoxyguanosine (8-OHdG) was formed from dG and isolated DNA by photosensitization with rose bengal (RB) and methylene blue (MB). 8-OHdG formed from dG was decomposed by the photosensitization with these dyes. Singlet oxygen was concerned with the formation and decomposition of 8-OHdG by photosensitization. Fe++ oxidized dG to 8-OHdG, which was decomposed by the addition of H2O2. 8-OHdG was formed and decomposed by the treatment of dG with Fe++, EDTA and ascorbic acid. Hydroxy radical (.OH) participated in the formation and decomposition of 8-OHdG by Fe++.

8-Hydroxy-2'-Deoxyguanosine↗

Triple radioisotope technique in etiologic evaluation of space-occupying lesions of the liver.

The findings from 43 patients with histologically proven focal liver disease examined by triple isotope tracer study are presented. The method consisted in a three step technique, the first step being an isotope scan after injection of 99mTc-sulfur colloid and 131I rose bengal. As a second step dynamic hepatic scintiangiography was carried out with 99mTc human serum albumin in order to assess the vascularity of a lesion detected on the initial scan. Thirdly, the patient was administered 67Ga to evaluate further the pathologic process. According to our results this triple radioisotope technique provides the clinician with valuable information on the possible etiology of focal liver disease.

Adenocarcinoma↗

Yeast phenylalanyl-tRNA synthetase. Properties of the histidyl residues.

Reactivity of the histidyl groups of yeast phenylalanyl-tRNA synthetase was studied in the absence or presence of substrates. In the absence of substrates about 10 histidine residues were found to react with similar kinetic constants. Phenylalanine at 10(-3) M was found to protect two histidyl residues; increasing the amino acid concentration to 5 . 10(-3) M resulted in the protection of two more histidyl groups. tRNAPhe did not afford any protection to histidine residues, but acylated phenylalanyl-tRNA (Phe-tRNAPhe) protected two of the four histidyl groups already protected by phenylalanine. These results suggest the existence of two different sets of accepting sites for phenylalanine: one specific for the free amino acid, the other one specific for the amino acid linked to the tRNA, but being accessible to free phenylalanine, with a somewhat lower binding constant, ATP was found to mask around four histidyl residues against diethylpyrocarbonate modification. By photoirradiation of enzyme-phenylalanine complex in the presence of rose bengale, a significant amount of amino acid was bound to the alpha subunit (Mr = 73 000) of phenylalanyl-tRNA synthetase, confirming that the amino acid binding site is located on this subunit, as previously suggested by modification of thiol groups. Upon irradiation of an enzyme-tRNA complex, almost no covalent binding of tRNA occurred during enzyme inactivation, suggesting that the histidyl residues involved in the enzymic activity are not required for tRNA binding.

Adenosine Triphosphate↗

A comparison of three serological tests in the diagnosis of caprine brucellosis.

The serum agglutination test (SAT), the Rose Bengal plate test (RBPT) and the milk ring test (MRT) were used in the diagnosis of caprine brucellosis. There was a close correlation between the SAT and RBPT when both tests were negative but the RBPT failed to detect 79.82 per cent of sera in excess of 50 iu. Also, owing to the relatively poor milking potential of the Nigerian goat and the false positive results with the MRT, it is concluded that the SAT offers a better serological diagnostic tool for caprine brucellosis in this locality.

Agglutination Tests↗

Effects of singlet oxygen on human lens crystallins in vitro.

We have recently demonstrated that singlet oxygen-mediated photooxidation can cause an increase in blue fluorescence and covalent cross-linking in bovine lens crystallins in vitro. Because these changes closely parallel modifications known to occur in human crystallins during aging and cataractogenesis, it was suggested that singlet oxygen may play an important role in these processes in vivo. to confirm these results, obtained with photosensitizers free in solution, we now report results from experiments with a polymer-bound photosensitizer and with photophysically generated singlet oxygen. Singlet oxygen photodynamically generated by polymer-immobilized rose bengal produced the following modifications to human lens crystallins in vitro: covalent cross-linking, increased blue fluorescence, yellow pigmentation, and formation of heavy-molecular-weight aggregates. Singlet oxygen generated photophysically was shown to cause an increase in blue fluorescence and covalent cross-linking in human crystallins. These findings substantiate the hypothesis that singlet oxygen may play an important role in the changes seen in human lens proteins during aging and cataractogenesis.

Aging↗

Effect of altered thyroid status on the transport of hepatobiliary radiopharmaceuticals.

The effect of induced hypothyroidism (by feeding an antithyroid drug-propylthiouracil) on the transport and clearance of the routinely used hepatobiliary radiopharmaceuticals--radioiodinated iodine-131 (131I) rose bengal and technetium-99m-N-(4-n-butylphenylcarbamoylmethyl) iminodiacetate, was studied in the rats. Hypothyroidism was associated with depressed growth and retarded clearance of these radiotracers from the in vivo system. Treatment of the hypothyroid rats with thyroxine (2-5 micrograms/100 g b.w. day) for 6 wk, restored these parameters towards normal values. These data suggest that delayed clearance of these hepatobiliary tracers could be related to reduced metabolic rate accompanied with the hypotonia and hypomotility of intestine normally observed in the hypothyroid state.

Animals↗

Photochemical relationships in Sacoglossan polypropionates.

Cyercene A has been synthesized and converted to placidene A and isoplacidene A through photoisomerization with sunlight. Hydroperoxide, isolated form Placida dendritica, was synthesized both by singlet oxygenation of cyercene A using Rose Bengal and through the irradiation of cyercene A alone in aerobic solution. The observation that hydroperoxide could be made by singlet oxygenation of cyercene A and that this occurs when cyercene A alone is irradiated supported the hypothesis that gamma-pyrones may act as triplet sensitizers. This was confirmed using cyercene A and a model gamma-pyrone to sensitize the photooxidation of n-butyl sulfide. The biosynthetic implications of these observations for Sacoglossan polypropionates are discussed.

Animals↗

Short-term effect of latanoprost and timolol eye drops on tear fluid and the ocular surface in patients with primary open-angle glaucoma and ocular hypertension.

PURPOSE: To investigate and compare the short term effects of topical latanoprost and timolol on the tear fluid and ocular surface condition in patients with bilateral primary open angle glaucoma or ocular hypertension. METHODS: Thirty-seven patients were included in this randomized, double-masked, parallel group study. Patients received either latanoprost 0.005% (n= 18) or timolol 0.5% (n= 19) instilled once daily in the morning for a treatment period of 27 days. Routine ophthalmic examinations, including intraocular pressure measurement, as well as tests to evaluate tear fluid and the ocular surface were performed. RESULTS: After one drop of medication, tear secretion was significantly reduced by timolol, but not by latanoprost. At the end of the study the break-up time (BUT) was significantly decreased in the timolol group but not in the latanoprost group. The BUT still remained in the normal range in both groups, although it is important to note that timolol was administered at half the clinical dose. Both latanoprost and timolol tended to increase Rose-Bengal staining of the cornea and conjunctiva after one month of treatment but no statistically significant difference was found between the groups. Corneal sensitivity was within the normal range for all patients during the study. CONCLUSION: Regarding ocular surface effects, no clinically important differences between latanoprost and timolol were observed as all the effects remained in the normal range.

Adrenergic beta-Antagonists↗

Activity loss and histidine modification in guanine deaminase.

Photooxidation of guanine deaminase (E.C.3.5.4.3.) in the presence of rose bengal as sensitizer resulted in decay in enzyme activity. The pH profile of inhibition suggests modification of histidyl residue(s). Amino acid analysis during photooxidation showed a decrease in unmodified hystidine. Diethylpyrocarbonate inhibits the enzyme activity whilst 5-aminoimidazole-4-carboxamide, a competitive inhibitor of guanine deaminase, partially protects against Et2PC action. Hydroxylamine partially reverses inhibition. These results are consistent with the presence of the imidazole moiety of the putative histidyl residue(s) at or near the active site of guanine deaminase.

Aminohydrolases↗

Prevalence of bovine and human brucellosis in western Algeria: comparison of screening tests.

A serological study was carried out in Tiaret province in western Algeria on 1032 cows distributed in 95 flocks to estimate the prevalence of Brucella infection and to compare the sensitivity and specificity of a range of agglutination tests. Screening tests showed 31.5% of herds positive using the buffered plate antigen test and 26.3% using the rose Bengal test compared with 15.7% with the complement fixation test. Using the complement fixation test as the gold standard for confirmatory tests, the Rivanol test was found to be more sensitive but less specific than tube agglutination in detecting brucellosis infection. Three isolates were identified from 105 blood samples from humans with brucellosis and 50 samples of milk and tissues from infected cows and they were all Brucella melitensis biovar 3.

Agglutination Tests↗