Vitamin C dose-dependently ameliorates renal hemodynamic toxicity of cisplatin in adult Swiss albino rats: a histopathologic and biochemical study

Nephrotoxicity is usually thought of as a common invariable consequence of hemodynamic toxicity whose effects, including oliguria and dysuria, has largely limited the clinical use of cisplatin. In this study, we investigated the protective effects of low and high dose of vitamin C against cisplatin-induced rat nephrotoxicity. Hence, 50 adult male Swiss albino rats were randomly divided into five equal groups to receive a corresponding dose of either normal saline as control, vitamin C (600 mg/kg/BW, i.v.), or cisplatin alone (7 mg/kg/BW, i.p.) or in combination with vitamin C at low dose (200 mg/kg/BW, i.v.) and high dose (600 mg/kg/BW, i.v.) for 9 days. Daily administration of cisplatin at a dose of 7 mg/kg/BW resulted in a significant increase in oxidative stress in renal tissues and plasma and a concomitant decrease in the creatinine clearance and renal blood flow as a result of early hemodynamic toxicity. Histopathological examination revealed acute tubular necrosis with hyaline cast formation triggered by cisplatin over 9 days of experiment. Further biochemical studies showed protecting effects of supplemented vitamin C at a high dose, illustrated by slowdown in the urinary enzyme activity, a significant decrease in plasma lipid peroxidation, and an increased tissue superoxide dismutase activity with recovery in the glomerular hemodynamicity and the ATPase activity up to 50 % when compared to controls and rats receiving low-dose. In high-dose animals, normal glomerular and tubular function on recovery from toxic renal failure led us to conclude that antioxidant property of vitamin C increases with dose, and, therefore, high dose of vitamin C prevents both functional and histological renal changes induced by cisplatin in rats, more efficient than low dose of the vitamin.     

A mouse model of argininosuccinic aciduria: biochemical characterization

Argininosuccinate lyase (AL) has several roles in intermediary metabolism. It is an essential component of the urea cycle, providing a pathway for the disposal of excess nitrogen in mammals. AL links the urea cycle to the tricarboxylic acid (TCA) cycle by generating fumarate. Finally, AL is required for the endogenous production of arginine. In this latter role it may function outside ureagenic organs to provide arginine as a substrate for nitric oxide synthases (NOS). Increasing evidence suggests that argininosuccinate synthetase (AS) and AL are more globally expressed, and the coordinate regulation of AS and AL gene expression with that of the inducible form of NOS (iNOS) provides evidence that this may facilitate the regulation of NOS activity. Deficiency of AL leads to the human urea cycle disorder argininosuccinic aciduria. We produced an AL deficient mouse by gene targeting in order to investigate the role of AL in endogenous arginine production. This mouse also provides a model of the human disorder to explore the pathogenesis of the disorder and possible new treatments. Metabolic studies of these mice demonstrated that they have the same biochemical phenotype as humans, with hyperammonemia, elevated plasma argininosuccinic acid and low plasma arginine. Plasma nitrites, derived from NO, were not reduced in AL deficient mice and there was no significant difference is the level of cyclic GMP, the second messenger induced by NO.     

Dose-related protecting effects of vitamin C in gentamicin-induced rat nephrotoxicity: a histopathologic and biochemical study

Gentamicin remains the mainstay in treatment of gram-negative infections, despite its potential ototoxicity and nephrotoxicity. In this study, we investigated dose-related protecting effects of vitamin C against gentamicin-induced rat nephrotoxicity. Hence, 50 male albino Wistar rats were randomly divided into five equal groups to receive a corresponding dose of either normal saline as control, vitamin C (200 mg/kg/bw, i.m.) or gentamicin alone (80 mg/kg/bw, i.m.) or in combination with vitamin C at low dose (200 mg/kg/bw, i.m.; LVG) and high dose (600 mg/kg/bw, i.m.) for 9 days. Daily administration of gentamicin at a dose of 80 mg/kg resulted in a significant increase in oxidative stress in renal tissues and plasma and a concomitant decrease in the creatinine clearance and renal blood flow as result of early hemodynamic toxicity. Histopathological examinations revealed acute tubular necrosis with hyaline cast formation triggered by gentamicin over 9 days of experiment, in addition to interstitial nephritis and tubular epithelial loss. Further biochemical studies showed protecting effects of supplemented vitamin C at a high dose, including slowdown in the urinary enzyme activity, a significant decrease in plasma lipid peroxidation, and an increased tissue superoxide dismutase activity with recovery in the glomerular hemodynamicity and the ATPase activity up to 50% when compared to controls and low-dose rats (LVG). In high-dose animals, normal glomerular and tubular function on recovery from toxic renal failure led us to conclude that antioxidant properties of vitamin C consistently increase with dose intensity. The present study also provided evidence that high dose of vitamin C prevented both functional and histological renal changes induced by gentamicin in rats, more efficient than low dose of the vitamin.     

Evaluation of a new cardiac recovery system in a bovine model of volume overload heart failure

We evaluated the short-term hemodynamic efficacy of the Cancion cardiac recovery system (CRS) in a bovine model of volume overload heart failure. We created severe mitral regurgitation (MR) by disrupting the mitral chordae tendineae of two calves, which were allowed to survive for 13 and 11 months. Four hours before we killed the calves, we introduced the CRS, which maintained flows of 1.2 L/min. Hemodynamic data were recorded before and after MR creation and CRS initiation. Left ventriculography was performed at those same intervals and 4 hours after CRS initiation. After chordal disruption, the left ventricular end-diastolic pressure (LVEDP) increased from 14 to 23 mm Hg in calf 1 and 15 to 27 mm Hg in calf 2. After 4 hours of CRS support, the LVEDP decreased from 23 to 8 mm Hg (calf 1) and 27 to 14 mm Hg (calf 2); dP/dt increased from 1,373 to 2,900 mm Hg/s (calf 1) and 1,068 to 2,384 mm Hg/s (calf 2). MR decreased from 3+ to 1 in calf 2 but could not be determined in calf 1. In this bovine model of volume overload heart failure, the CRS unloaded the left ventricle and reduced the afterload. Future trials will determine the pump's ability to treat congestive heart failure.     

Molecular and biochemical characterization of a Coccidioides immitis-specific antigen.

Results of earlier investigations have indicated that the saprobic phase of Coccidioides immitis produces a heat-stable, 19-kDa antigen with serine proteinase activity which has been suggested to be specific for this pathogenic fungus. In the present study we have determined the N-terminal and partial internal amino acid sequences of the purified, 19-kDa antigen, cloned the gene which encodes this polypeptide, and confirmed that the secreted proteinase is a Coccidioides-specific antigen (CS-Ag). Both the genomic and cDNA sequences are reported and reveal that the csa gene which encodes this antigen has no introns. A 543-bp open reading frame encodes a 181-amino-acid-containing protein with a predicted molecular mass of 19.8 kDa and an isoelectric point of 8.3. The csa gene was localized on chromosome I of three representative C. immitis clinical isolates on the basis of Southern hybridizations. Expression of the csa gene in Escherichia coli using the pET21a plasmid vector yielded a recombinant protein that was recognized in immunoblot assays by antibody raised to the purified 19-kDa CS-Ag. Secretion of the native antigen is suggested to occur by cleavage of a putative 23-residue signal peptide. The native CS-Ag showed a low degree of glycosylation. Analysis of the carbohydrate composition of the CS-Ag revealed xylose, mannose, galactose, and glucose. However, the purified antigen showed no affinity for concanavalin A. A PCR method with specificity and high sensitivity for detection of C. immitis genomic DNA, using a pair of synthetic oligonucleotide primers whose sequences were based on that of the csa gene, was developed. A 520-bp product was amplified only when C. immitis genomic DNA was used as the template. The lower limits of DNA detection using this PCR method were 1 pg of C. immitis genomic DNA by ethidium bromide staining and 100 fg after Southern hybridization. The csa gene-based PCR method for detection of C. immitis DNA is useful for culture identification and may have clinical applications for the diagnosis of coccidioidal infections.     

Vitamin B6 dose-dependently ameliorates renal hemodynamic toxicity of cisplatin in rat model of nephrotoxicity: the histopathologic and biochemical findings

Despite its significant anticancer activity, the clinical use of cisplatin is often limited by its undesirable side effects in the kidney known as nephrotoxicity. It is a common and often overlooked clinical entity that presents itself in the setting of oxidative stress-associated diseases in older individuals with renal failure. In this study, we investigated the antioxidant-protecting effects of vitamin B₆ in the kidney, with a view on the vasoregulatory role of renal pyridoxal 5′-phosphate at reducing the hemodynamic toxicity of cisplatin. Hence, 50 male Sprague–Dawley rats were randomly assigned in one of five groups of the study to receive a corresponding dose of either normal saline, vitamin B₆ (200 mg/kg/bw; i.m.) or cisplatin alone (7 mg/kg/bw; i.m.), or in combination with vitamin B₆ at low (100 mg/kg/bw; i.m.) and high dose (200 mg/kg/bw; i.m.) for 10 days as animal model of renal failure. Daily administration of cisplatin at a dose of 7 mg/kg/bw resulted in a significant increase in local and systemic oxidative stress of the kidney and a decrease in glomerular function as a result of early hemodynamic toxicity. Histopathological examinations of renal tissues revealed acute tubular necrosis with hyaline cast formation triggered by cisplatin over 9 days of the study, in addition to interstitial nephritis and tubular epithelial loss. Further biochemical studies in HVB group showed the protecting effects of supplemented vitamin B₆ at a high dose, including a slowdown in urinary enzyme activity, a significant decrease in plasma lipid peroxidation, and an increased tissue superoxide dismutase activity with recovery in the glomerular hemodynamicity and ATPase activity up to 50 % when compared to the low-dose rats and controls. In high-dose animals, the normal glomerular and tubular function on recovery from toxic renal failure led us to conclude that the antioxidant property of vitamin B₆ consistently increases with the dose intensity. The present study also provided evidence that high dose of vitamin B₆ prevented both functional and histological renal changes induced by cisplatin in rats, more efficient than low dose of the vitamin.     

Streptococcus salivarius urease: genetic and biochemical characterization and expression in a dental plaque streptococcus.

The hydrolysis of urea by urease enzyme of oral bacteria is believed to have a major impact on oral microbial ecology and to be intimately involved in oral health and diseases. To begin to understand the biochemistry and genetics of oral ureolysis, a study of the urease of Streptococcus salivarius, a highly ureolytic organism which is present in large numbers on the soft tissues of the oral cavity, has been initiated. By using as a probe a 0.6-kpb internal fragment of the S. salivarius 57.I ureC gene, two clones from subgenomic libraries of S. salivarius 57.I in an Escherichia coli plasmid vector were identified. Nucleotide sequence analysis revealed the presence of one partial and six complete open reading frames which were most homologous to ureIAB-CEFGD of other ureolytic bacteria. Plasmid clones were generated to construct a complete gene cluster and used to transform E. coli and Streptococcus gordonii DL1, a nonureolytic, dental plaque microorganism. The recombinant organisms expressed high levels of urease activity when the growth medium was supplemented with NiCl2. The urease enzyme was purified from E. coli, and its biochemical properties were compared with those of the urease produced by S. salivarius and those of the urease produced by S. gordonii carrying the plasmid-borne ure genes. In all cases, the enzyme had a Km of 3.5 to 4.1 mM, a pH optimum near 7.0, and a temperature optimum near 60 degrees C. S. gordonii carrying the urease genes was then demonstrated to have a significant capacity to temper glycolytic acidification in vitro in the presence of concentrations of urea commonly found in the oral cavity. The ability to genetically engineer plaque bacteria that can modulate environmental pH through ureolysis will open the way to using recombinant ureolytic organisms to test hypotheses regarding the role of oral ureolysis in dental caries, calculus formation, and periodontal diseases. Such recombinant organisms may eventually prove useful for controlling dental caries by replacement therapy.     

Collaborative care: a new model for a new century.

From the imagined vantage point of the year 2020, the author recounts the problems and deficiencies of health care in the 1990s and describes how academic medicine's leaders successfully confronted them. A key part of their strategy was to work together to form a coordinated network of medical schools, teaching hospitals, and academically oriented health systems, along with their staffs and a variety of community-based partners. In this way, they set a national agenda, pursued common goals, freely shared information and best practices, and cooperated to optimize their effectiveness in education, research, and clinical care. A major outcome of this new network was the Collaborative Care model of health care, based on the premise that a basic purpose of the health care system is to achieve measurable improvements in the health of individuals and communities in ways that are cost-conscious, quality-driven, evidence-based, and patient-, family-, and community-oriented. Academic institutions formed strong partnerships with many stakeholders (e.g., purchasers of health care services) to make the Collaborative Care approach work. In addition, there were several other important keys to Collaborative Care's success, such as the full integration of clinical research with clinical care and the restoration of trust in the health care enterprise. The author returns to reality and the 1990s. He challenges academic medicine to pursue the Collaborative Care vision, saying that "we should not accept without challenge what we know to be abominable just because it appears to be inevitable.... Our choice is to continue to struggle for survival as the environment around us gets harsher and harsher ... or to fix the environment" using the power of collaboration to unleash academic medicine's unlimited creativity and wisdom.     

Brain dialysis: detection of acetylcholine in the striatum of unrestrained and unanesthetized rats

A dialysis cannula was implanted into rat striatum while the animals were anesthetized, and after at least one day following the surgery the area was perfused with Ringer solution under the unrestrained and unanesthetized conditions. Concentration of acetylcholine (ACh) in the perfusate was determined by high-performance liquid chromatography (HPLC)-electrochemical detection (ECD) with the enzyme-column on which acetylcholine esterase and choline oxidase were immobilized. ACh in the dialysate was only detectable when the Ringer solution containing eserine, an inhibitor of acetylcholinesterase, was perfused. ACh peak on HPLC-ECD could be detected at least for 4 h under these conditions. The level of ACh increased 2-3 fold with the perfusion of 1 mM atropine sulfate, a blocker of ACh receptor. These data indicate that brain dialysis in the presence of eserine is useful for study on the neurochemical activity of ACh neurons in the brain.     

Continuous measurement of cerebrospinal fluid pressure in unrestrained rats

A sensitive method for measuring intracranial pressure in unrestrained, unanesthetized rats has been developed. Two identically calibrated pressure transducers connected to fluid-filled polyethylene catheters were used to simultaneously monitor ventricular cerebrospinal fluid (CSF) pressure and head position. Artifacts due to changing head position were electronically subtracted from ventricular pressure, yielding a continuous dynamic record of “true” CSF pressure. A simple technique for precisely calibrating pressure transducers is also described. The mean CSF pressure obtained in 27 non-stressed freely-behaving adult rats was 64 ± 2 mm H₂O. Handling caused a prompt increase in intracranial pressure of approximately 80–120 mm H₂O. Moment-to-moment fluctuations in intracranial pressure closely reflect changes in central venous pressure. The techniques described are applicable to continuous measurement of CSF pressure in laboratory or clinical settings.     

Comparative study between biochemical and histological methods and image analysis in liver iron overload.

Iron overload has been measured in 100 hepatic biopsies by three different methods: (i) biochemical assay of the liver iron concentration (LIC), (ii) histological grading (HISTO) and (iii) automated image analysis with a Leitz Texture Analysis System by estimating two parameters, (a) the total iron area (TIA) and (b) the first grey level step (FGLS) at which the iron is detected. Image analysis appears as a specific, sensitive, quick, reproducible and valid method.     

Effect of morphine and naloxone on acupuncture analgesia in unrestrained rats

Experiments on unrestrained rats showed that electroacupuncture of an acupuncture point distinctly reduces the nociceptive response to electrical stimulation of the base of the tail. Morphine, in a subanalgesic dose (5 mg/kg), potentiated the analgesic effect of acupuncture. Naloxone, in a dose of 5 mg/kg, completely abolished acupuncture analgesia. The possible mechanisms of analgesia production by electroacupuncture are discussed.Department of Pharmacology, I. P. Pavlov First Leningrad Medical Institute. Course of Reflex Therapy, S. M. Kirov Leningrad Postgraduate Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR V. V. Zakusov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 11, pp. 566–569, November, 1979.     

Heart functional responses to pressure overload in exercised and sedentary rats.

Female rats that had been subjected to a moderate treadmill running program were compared with sedentary animals on the basis of heart weight, selected biochemical measurements, and heart function. Exercised animals maintained normal growth rate, and cardiac hypertrophy was not present. Left ventricular RNA, DNA, and cytochrome c levels were unchanged. Heart functional measurements obtained in situ were similar in sedentary and exercised animals under control conditions. When subjected to sustained (1-3 days) aortic constriction pressure overload, exercised animals maintained or increased myocardial contractility. Contractility was depressed in sedentary animals. Both sedentary and exercised animals increased left ventricular end diastolic pressure without changing contractility during acute (1-3 min) pressure overload. However, exercised animals were able to fully regain normal cardiac output when the acute overload was relieved. Cardiac output remained approximately 10% below control in sedentary animals. The improved ability of previously exercised animals to withstand pressure overload appears to be due to alterations in adaptation rather than preliminary augmentation of metabolism or function.