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Biomedical subjects

G Rao

Publications and source records attributed to G Rao.

At least 91 records · Page 5Linked to original sources
PubMed1995

Low levels of reactive oxygen species as modulators of cell function.

In this paper, we present various arguments supporting the hypothesis that reactive oxygen species (ROS) could be responsible for the modulation of various cellular functions, besides their well known toxic effects. We first review the recent evidence indicating that ROS are able to modulate genome expression through specific and precise mechanisms during cell activation. The role of the nitrogen reactive radicals such as nitric oxide is separately analyzed because of its specific role in the nervous and vascular systems. The action of the other ROS on gene activation will then be reviewed by first looking at their possible involvement in the activation of transcription factors like NF-kappa B. Arguments will then be developed in favor of the implication of the ROS in the cellular effects of PMA, TNF-alpha and other cytokines on the modulation of the genetic expression. Possible mechanisms will be presented for linking the production of the ROS with cell activation. In a general way we postulate that ROS can play a role of secondary messengers in several cell responses to external stimuli. In the second part of the paper, we will examine the long term influence of ROS and their possible roles in cellular aging. Different links exist between ROS and aging and the relationship between them is probably indirect. We propose to consider the effect of ROS as one of the multiple challenges that cells have to face, the cell being considered as a global system which must optimize its energy expenditure for carrying out its basic functions such as turnover, differentiated phenotype functions, multiplication, defense and repair processes. This thermodynamic point of view will help to understand the effect of low ROS stresses, among others, on accelerated aging.

Animals↗
PubMed1995

Sensing oxygen through skin using a red diode laser and fluorescence lifetimes.

The most difficult impediments to transcutaneous optical sensing are the absorbance and scatter of light caused by skin and the lack of fluorescent sensing probes which can be excited at wavelengths over 600 nm. Furthermore, current optical sensing techniques rely on absorbance or fluorescence intensity measurements, both of which are sensitive to drifts in lamp intensity, changes in probe concentration and inner filter effects. We demonstrate oxygen sensing through a layer of skin by using red light which readily penetrates skin as diffusely scattered light. The oxygen sensitive osmium-ligand complex used in this study can be excited at 635-680 nm. In addition, we measure fluorescence lifetimes, which are inherently unaffected by factors that limit absorbance and fluorescence intensity measurements. By using phase fluorimetry and long lived fluorophores, we are able to demonstrate the potential for subdermal oxygen sensing with simple and inexpensive instrumentation. This work describes a paradigm for future non-invasive measurements of other analytes.

Biosensing Techniques↗
PubMed1995

Potential applications of lifetime-based, phase-modulation fluorimetry in bioprocess and clinical monitoring.

The measurement of analyte concentration is a critical part of successful bioreactor and clinical monitoring. Although strategies exist for measuring the majority of relevant analytes, industrial on-line bioreactor control is carried out primarily by measurement and control of pH, pO2 and, in some cases, cell density. This is because the available technology cannot be easily and inexpensively adapted to (a) measure the analyte in an aseptic manner and/or allow for remote sensing, and (b) measure in real time so that on-line control is possible. Similar issues need to be addressed for biosensors for clinical applications. A rapidly emerging technology that has the potential of meeting these challenges is liftime-based phase-modulation fluorimetry, an optical technique that uses the measurement of fluorescence lifetime rather than intensity for determining the concentration of an analyte.

Biosensing Techniques↗
PubMed1995

Reverse iontophoresis: noninvasive glucose monitoring in vivo in humans.

PURPOSE: To demonstrate that "reverse iontophoresis" can be used to noninvasively obtain information about systemic glucose levels in vivo in humans. METHODS: The passage of current across the skin in vivo drives ions into the tissue, from the electrode chambers positioned on the skin surface, and simultaneously pulls ions from the body in the opposite direction. Because of the net negative charge on the skin, under normal conditions, the membrane is permselective to cations, and a potential gradient also results, therefore, in electroosmotic convection of solvent in the direction of counterion flow (i.e., from anode to cathode). Thus, it is also possible to enhance the transport of polar, yet uncharged, species using iontophoresis. In an earlier study, the in vitro extraction of glucose, by "reverse iontophoresis" was established, and extension of the approach to an in vivo model was indicated. The idea has therefore been further explored in vivo in humans. RESULTS: Using small, simple, prototypical electrode chambers, attached to the ventral forearm surface, direct current iontophoresis at 0.25 mA/cm2 for periods of up to 1 hour, and a sensitive analytical procedure to measure the quantities of glucose extracted, it has been shown that iontophoretic sampling of glucose is feasible. However, the shorter periods (15 minutes or less) of extraction considered yield results which are "contaminated" (it is believed) by glucose that is a product of lipid metabolism within the skin. While this material is expected to complicate the initial calibration of the approach, the problem is effectively resolved within one hour, by which time the glucose arriving in the electrode chambers on the skin surface is expected to directly reflect the subcutaneous tissue concentration. CONCLUSIONS: Based upon these initial observations, further investigation can now be directed towards optimization of electroosmotic flow and sampling time, improved reproducibility and the development of a practical assay methodology.

Blood Glucose↗
PubMed1995

Activities of antioxidant enzymes in various tissues of male Fischer 344 rats are altered by food restriction.

The objective of this study was to determine how food restriction (40% restriction of food intake) altered the age-related changes in the activities of Cu,Zn superoxide dismutase, catalase and glutathione peroxidase in liver, brain cortex, heart, kidney and intestinal mucosa obtained from 6-, 16- and 26-mo-old male Fischer 344 rats. Food restriction increased the activity of one or more of the antioxidant enzymes in the liver, brain cortex, heart and kidney of the rats. However, the magnitude of the effect and the antioxidant enzyme(s) affected by food restriction varied from tissue to tissue, and food restriction had no significant effect on the activities of these enzymes in intestinal mucosa. Interestingly, the four tissues in which food restriction increased the activity of one or more of the antioxidant enzymes showed reduced lipid peroxidation as measured by thiobarbituric acid-reactive material. These data suggest that food restriction might enhance the survival of rodents by altering the levels of the antioxidant enzymes and hence reducing free radical damage.

Aging↗
PubMed1995

Hereditary spastic paraplegia with dementia.

OBJECTIVE: Hereditary spastic paraplegia (HSP) with dementia is a very rare condition. The aim of the paper is to present the first report of HSP in a Fijian Indian family. METHOD: A psychiatrist and a general physician examined the affected members of the family on five occasions over three years. RESULTS: There are three affected individuals in a sibship of seven. The parents are without symptoms and the marriage is non-consanguineous. The course of the disease has been remarkably similar. All subjects were healthy and performing well in the early years of school. In two, symptoms of cognitive loss preceded difficulty with ambulation and in the third, these symptoms appeared concurrently. All subjects had both symptoms by 13 years of age; they were unable to ambulate independently by the mid to late teens, at which time there was dysarthria spastic paraplegia and dementia. One subject suffered a three month episode of hypomanic behaviour. Over the three-year study period deterioration was slight but noticeable. CONCLUSIONS: It is possible that HSP is more commonly associated with pre-senile dementia than is currently recognised. HSP with dementia is a very rare cause of failing school performance. Physical examination of the patient and other family members is indicated if this diagnosis is being considered.

Adolescent↗
PubMed1995

[A study on the simultaneous HPLC determination of chlorhexidine and its impurity 4-chloroaniline].

This paper reports a new RP-HPLC method using phenacetin internal standard (IS) for simultaneous determination of chlorhexidine (CH) and its impurity 4-chloroaniline (4-CA). We use the LC-3A HPLC instrument, a Zorbax-C8 column (25cm x 4.6mm id, 10 microns), a variable wave length UV detector and C-RIA chromatographic data processor. A mixture of methanol and 0.2 mol/L NaH2PO4 (50.5 : 49.5, pH 3.0) is used as mobile phase, whose flow rate is 1.0 ml/min. Sample solution of 15 microliters is injected onto the column, which is maintained at 30 degrees C. The eluent is monitored at 240nm. The retention times are 16.72 min for CH and 5.54 min for 4-CA. The standard curves of CH and 4-CA are established using peak area ratio of CH/IS and 4-CA/IS to concentration respectively. They are both linear (r = 0.9999) within 15-200 micrograms/ml for CH and 60-1200 micrograms/ml for 4-CH. The limits of detection are 1.33 micrograms/ml for CH AND 7.5 micrograms/ml for 4-CA (S/N = 3). The analytical recovery rates are 100.1% for CH and 103.1% for 4-CA. This method has been used for investigating the stability of CH preparation.

Aniline Compounds↗
PubMed1994

Persistent increase of hippocampal presynaptic axon excitability after repetitive electrical stimulation: dependence on N-methyl-D-aspartate receptor activity, nitric-oxide synthase, and temperature.

The electrical excitability of Schaffer collateral axons and/or terminals was studied in hippocampal slices by monitoring single, CA3 pyramidal neurons activated antidromically from CA1 stratum radiatum. At 22 degrees C, weak, repetitive stimulation with as few as 10 impulses at 2 Hz led to a robust lowering of the antidromic activation threshold that lasted > 30 min. The effect was completely absent at 32 degrees C and was blocked by both the N-methyl-D-aspartate receptor antagonist, 2-amino-5-phosphonovalerate and the inhibitor of nitric-oxide synthase, L-nitro-arginine methyl ester. Such threshold lowering would alter the variance of synaptic responses from axons stimulated in the variable excitation region of their input-output functions. These results thus raise important doubts about the interpretation of experiments in which the so-called minimal-stimulation method has been used at reduced temperature to infer changes in quantal transmission during hippocampal long-term potentiation. In the present experiments, no changes were observed in the estimate of excitatory postsynaptic potential quantal content in long-term potentiation experiments at either temperature, which could not be accounted for by an artificial, temperature-dependent change in the responsiveness of presynaptic axons.

Amino Acid Oxidoreductases↗
PubMed1994

tert.-butyl hydroperoxide-mediated DNA base damage in cultured mammalian cells.

tert.-Butyl hydroperoxide has been utilized to study the effect of oxidative stress on living cells; however, its effect on DNA bases in cells has not been characterized. In the present work, we have investigated DNA base damage in mammalian cells exposed to this organic hydroperoxide. SP2/0 derived murine hybridoma cells were treated with 4 concentrations of tert.-butyl hydroperoxide for varying periods of time. Chromatin was isolated from treated and control cells and subsequently analyzed by gas chromatography-mass spectrometry with selected-ion monitoring for DNA base damage. Quantification of damaged DNA bases was achieved by isotope-dilution mass spectrometry. The amounts of 8 products were significantly higher than control levels in cells treated with tert.-butyl hydroperoxide at a concentration range of 0.01-0.1 mM. At concentrations from 1.0 to 10 mM, product formation was inhibited and the amounts of products were similar to those in control cells. The bimodal nature of the dose-response may be qualitatively analogous to previous reports of bimodal killing of E. coli bacteria by hydrogen peroxide. The nature of the identified DNA base lesions suggests the involvement of the hydroxyl radical in their formation. tert.-Butyl hydroperoxide is known to produce the tert.-butoxyl radical in reactions with metal ions. However, it is unlikely that the tert.-butoxyl radical produces these DNA lesions. It is suggested that DNA base damage arises from tert.-butyl hydroperoxide-mediated oxidative stress in cells, resulting in formation of hydroxyl radicals in close proximity to DNA. The inhibition of product formation at high concentrations of tert.-butyl hydroperoxide may be explained by the scavenging of tert.-butoxyl radical by tert.-butyl hydroperoxide resulting in inhibition of oxidative stress. The plausibility of the scavenging mechanism was evaluated with a mathematical simulation of the dose-response for DNA damage in solutions containing hydrogen peroxide. The simulation model predicted a bimodal dose-response which agreed qualitatively with the results in this study and with other in vivo and in vitro studies reported in the literature.

Animals↗
PubMed1994

Complexation of iron cation by sodium urate crystals and gouty inflammation.

Gouty inflammation can be suppressed by an iron chelator. We therefore hypothesized that arthritis associated with sodium urate crystal deposition could follow the incomplete complexation of iron cation with subsequent oxidant generation as the metal cycles through reduced and oxidized states. Urate crystals adsorbed Fe3+ in vitro and crystals collected from a human tophus had significant concentrations of ionizable iron. Urate crystals oxidized deoxyribose to a thiobarbituric acid (TBA)-reactive product, augmented luminol chemiluminescence by neutrophils, released leukotriene B4 from neutrophils, activated complement, and promoted neutrophil chemotaxis. All of these events increased with the concentration of complexed iron and were suppressed by the iron chelator deferoxamine or the .OH scavenger dimethylthiourea. These results suggest that some portion of gouty inflammation after urate crystal deposition could result from the incomplete complexation of iron with subsequent catalytic generation of reactive oxygen species.

Chemotaxis, Leukocyte↗
PubMed1994

Early complications of contemporary anatomical radical retropubic prostatectomy.

Anatomical radical retropubic prostatectomy is commonly performed to treat select patients with clinically localized prostate cancer. Because this recently introduced surgical technique is technically demanding, and since early detection programs have altered the age and stage distribution of men undergoing radical prostatectomy, earlier reports describing complication and death rates from radical prostatectomy may not accurately reflect contemporary experience. A retrospective review of 1,342 radical retropubic prostatectomies performed during a 5-year period showed an early complication rate of 7.4%. The most common complications were thromboembolic (pulmonary embolism) and cardiac (myocardial infarction). Patient age, mode of diagnosis (transurethral resection of the prostate versus needle biopsy) and pathological stage of the tumor did not correlate with the rate of complications. Three patients died in this series (0.2%). This retrospective review shows that contemporary anatomical retropubic radical prostatectomy may be performed with acceptably low early morbidity and mortality rates, and that appropriate selection of chronologically older patients is not associated with significantly greater rates of early complications. These data may be useful in decision analytic models evaluating the role of therapy for patients with early stage prostate cancer.

Adult↗
PubMed1994

Prevention of photocarcinogenesis and UV-induced immunosuppression in mice by topical tannic acid.

Topical application of tannic acid, a phenolic antioxidant derived from plants, was found to inhibit the cutaneous carcinogenesis and the immunosuppression induced by ultraviolet B (UVB) irradiation with no visible toxicity. BALB/cAnNTacfBR mice were treated with 200 micrograms of tannic acid three times weekly for two weeks before UV treatments began and throughout the experiment. UVB irradiation consisted of five 30-minute exposures per week to banks of six FS40 Westinghouse sunlamps. In the photocarcinogenesis study, mice received a total dose of approximately 1.09 x 10(6) J/m2. Skin cancer incidence in UV-irradiated mice was 75% at 26 weeks after the first UV exposure; tannic acid reduced this to 42%. Immunosuppression induced by UVB irradiation normally prevents the host from rejecting antigenic syngeneic UV-induced tumors. Immunosuppression in these experiments was measured by a passive transfer assay. Tumor challenges grew to an average of 88 +/- 20, 36 +/- 11, and 20 +/- 8 mm2 in naive recipients of splenocytes from UVB-irradiated mice, nonirradiated control mice, and UVB-irradiated mice treated with tannic acid, respectively. Thus topical tannic acid treatment prevented the transfer of enhanced tumor susceptibility with splenocytes from UVB-irradiated mice.

Administration, Topical↗
PubMed1993

Hyperoxia induces DNA damage in mammalian cells.

There is mounting evidence on the role of oxygen-derived free radicals in causing damage to various cellular components. However, most studies reported in the literature have been conducted under conditions where cells were challenged with chemical free radical generating systems. In contrast, we measured DNA strand breaks, through a relatively simple and sensitive technique, as a function of the dissolved oxygen tension in a bioreactor. Cells were exposed to a step change in oxygen tension at mid-exponential growth phase. Several levels of oxygen were tested (200, 300, and 476% dissolved oxygen with respect to air saturation at 1 atmosphere) and compared against a control (10% dissolved oxygen). Hyperoxia was found to cause monotonically increasing DNA strand breakage at all the oxygen levels. In addition, hyperoxia was found to affect other metabolic functions such as the glucose consumption rate, lactate production rate, and cell growth. When hyperoxia-induced DNA strand breakage was compared to that induced by exposure to hydrogen peroxide, a similar response was observed. Exposure to a dissolved oxygen level of 200% induced DNA strand breakage comparable to a bolus of 4.2 microM hydrogen peroxide. Our results show that there is an association between hyperoxia and DNA damage.

Animals↗