Methylated niacin derivatives in plasma and urine after an oral dose of nicotinamide given to subjects fed a low-methionine diet.

Five healthy males, age 25-32 y, were fed in sequence a diet of ordinary foods (10 d, PI), a low-methionine diet (285 mg/d, 14 d, PII), and an adequate-methionine diet (725 mg/d, 7 d, PIII). Diets contained 9 g nitrogen (N) per day with soy protein and synthetic L-amino acids as the N sources in PII and PIII. In PII, subjects were in negative N balance whereas, in PIII, four subjects were in positive N balance. On the last day of each period, fasting subjects ingested a dose of nicotinamide (NAM, 102 mumol/kg body wt). Plasma and urine samples were analyzed for methylated derivatives of NAM by high-performance liquid chromatography (HPLC) methods. Mean values of methylated metabolites in urine from the three diet periods (for four subjects in N balance during PIII) were not different (59.8, 56.7, and 59.9 mumol/(kg body wt X 24 h) for PI, PII, and PIII, respectively). Plasma values of these metabolites also were similar. Results suggest that during a 2-wk period of negative N balance due to a low-methionine intake hepatic methylation processes are not impaired. These processes appear to have a higher metabolic priority than maintenance of the net protein synthesis rate.

Indomethacin lowers the threshold thermal exposure for hyperthermic radiosensitization and heat-shock inhibition of ionizing radiation-induced activation of NF-kappaB.

PURPOSE: It is well established that salicylate and several other non-steroidal anti-inflammatory agents (NSAID), including indomethacin, can activate the heat-shock response, albeit at high concentrations. This is significant since heat shock significantly alters the cellular cytotoxic response to ionizing radiation (IR). It was previously shown that heat shock, as well as NSAIDs, inhibits IR-induced activation of NF-kappaB and that NF-kappaB protects against IR-induced cytotoxicity. Hence, it is hypothesized that pretreatment with indomethacin before heating will lower the temperature and heating times required to inhibit the activation of NF-kappaB and induce significant hyperthermic radiosensitization. MATERIALS AND METHODS: Experiments were performed in HeLa cell lines and the DNA-binding activity was determined by EMSA. Cellular radiosensitivity was determined by clonogenic assay. RESULTS: HeLa cells pretreated with indomethacin showed a decrease in the temperature-time combination necessary to inhibit IR-induction of NF-kappaB DNA binding. In addition, clonogenic cell survival assays using identical conditions showed an indomethacin dose-dependent enhancement of hyperthermic radiosensitization. Thus, similar concentrations of indomethacin both lowered the threshold thermal exposure to inhibit activation of NF-kappaB DNA-binding and increased the sensitivity of tumour cells to hyperthermic radiosensitization-induced cytotoxicity. In HeLa cells treated with N-alpha-tosylphenylalanyl-chloromethyl ketone (TPCK), a serine protease inhibitor that blocks activation of NF-kappaB, an increase in radiosensitivity was observed. Interestingly, no additional cell killing was observed when heat shock was added to cells treated with TPCK before IR, suggesting a possible common cytotoxic pathway. CONCLUSIONS: The results demonstrate that indomethacin lowers the temperature-time conbination necessary to induce several physiological processes associated with the heat-shock response. Furthermore, NSAID may be potential adjuvants in improving the clinical effectiveness of hyperthermia in radiation therapy.

N.Y. Hospital tests productivity model.

An operating room project with a goal of $1.5 million in savings was undertaken by New York Hospital, an academic medical center in New York City. The ORs were chosen because of their logistic and operational problems, but also because they serve as a microcosm of the hospital. An OR model could be integrated into overall productivity efforts.

Mineral balance and blood pressure in the young spontaneously hypertensive rat.

To evaluate factors that might be related to the pathogenesis of hypertension in spontaneously hypertensive rats (SHR), mineral balance studies were conducted in SHR and control Wistar-Kyoto (WKY) rats at the ages of 9, 13 and 16 wk. During the first balance period, before onset of hypertension, Na, Ca, Mg and P balances were all significantly more positive in SHR than in WKY rats. Percentage of absorbed Na excreted in urine was correlated with the rise in blood pressure of SHR but not of WKY rats during the first 4 wk of the study. At 13 wk balances of all four nutrients still tended to be higher in SHR. During the 16th wk of life, after onset of hypertension, Na, Ca and P balances were similar in SHR and WKY rats, but Mg balance was significantly lower in SHR. The percent urinary excretion of absorbed Na was lower in SHR at 9 wk of age, but after onset of hypertension it was similar to that in WKY rats. Food intake was always greater in SHR, whereas growth during the study was less, and absorption of Ca, Mg and P declined more rapidly with age in SHR than in WKY rats. The data suggest that nutrient malabsorption in SHR, if it occurs at all, is most likely an effect of, rather than a cause of, hypertension.

Probiotic spectra of lactic acid bacteria (LAB).

Lactic acid bacteria (LAB) and their probio-active cellular substances exert many beneficial effects in the gastrointestinal tract. LAB prevent adherence, establishment, and replication of several enteric mucosal pathogens through several antimicrobial mechanisms. LAB also release various enzymes into the intestinal lumen and exert potential synergistic effects on digestion and alleviate symptoms of intestinal malabsoption. Consumption of LAB fermented dairy products with LAB may elicit antitumor effects. These effects are attributed to the inhibition of mutagenic activity; decrease in several enzymes implicated in the generation of carcinogens, mutagens, or tumor-promoting agents; suppression of tumors; and the epidemiology correlating dietary regimes and cancer. Specific cellular components in LAB strains seem to induce strong adjuvant effects including modulation of cell-mediated immune responses, activation of reticuloendothelial system, augmentation of cytokine pathways and regulation of interleukins, and tumor necrosis factors. Oral administration of LAB is well tolerated and proven to be safe in 143 human clinical trials and no adverse effects were reported in any of the total 7,526 subjects studied during 1961-1998. In an effort to decrease the reliance on synthetic antimicrobials and control the emerging immunocompromised host population, the time has come to carefully explore the prophylactic and therapeutic applications of probiotic LAB.

Metabolic effects of histidine-deficient diets fed to growing rats by gastric tube.

Effects of histidine deficiency on muscle carnosine and anserine levels and on activities of enzymes associated with histidine catabolism and protoporphyrin synthesis were investigated. Male Sprague-Dawley (150 g) rats were tube-fed isonitrogenous, isocaloric, defined diets containing 0%, low (0.013%) or adequate (0.45%) histidine for 8-13 days. While histidine-deficient animals maintained body weight, muscle and plasma histidine and carnosine concentrations decreased rapidly and remained low following a 3-day histidine repletion period. Hepatic histidine ammonia-lyase and histidine-pyruvate transaminase activities were decreased in histidine-deficient animals, whereas formiminotransferase activity was unchanged. Hematocrit levels and hemoglobin concentrations declined progressively during histidine depletion and the activity of erythrocyte and hepatic delta-aminolevulinic acid dehydratase also decreased relative to controls. Evidence is presented indicating that decreased histidine catabolism combined with carnosine and hemoglobin degradation can provide sufficient histidine to explain the slow onset of negative nitrogen balance associated with histidine deficiency and that impaired protoporphyrin synthesis may partially explain the anemia observed in the absence of dietary histidine.

Hypotensive effect of a high calcium diet in the Wistar rat.

In order to study the effects of Ca supplementation upon systolic blood pressure, male weanling Wistar rats were fed rat chow or chow mixed with either CaCO3 (to 2.5% total Ca) or CaHPO4 (to 3.4% total Ca). In the first study rats receiving CaCO3 supplementation had lower systolic pressures after 6 weeks on the diet. Urinary P was reduced, fecal P was increased and urinary Ca was increased with this diet. Ionized Ca in blood was unaltered by Ca supplementation, but total Ca was decreased and plasma P was increased. In the second study with CaCO3, only a transient effect on blood pressure was found. The effects of the diet upon Ca and P metabolism, however, were similar to those found in the first study. In the third study, with CaHPO4 as the vehicle for Ca supplementation, blood pressure was consistently reduced in rats after 8 weeks on the high Ca diet. Plasma P, total Ca and whole blood ionized Ca were unaltered by this diet, but ionized Ca was reduced in plasma. The dietary intake of nutrients other than the Ca salt did not differ between control and and experimental groups in any of the experiments. Urinary Na and K were similar in control and experimental animals in all three experiments. These data suggest that neither increased ionized Ca, P depletion nor increased Na excretion are necessary conditions for expression of the hypotensive effect of supplementary Ca.

Comparison of calcium and magnesium absorption: in vivo and in vitro studies.

Transport characteristics of Ca and Mg were compared at three different levels of the absorptive process in male (120-170 g) Wistar rats. Balance studies in intact rats revealed that fractional Ca absorption decreased with increased Ca intake so that net Ca absorption remained constant. Fractional Mg absorption decreased modestly with increased Mg intake so that net Mg absorption increased proportionately with increased dietary Mg. Everted duodenal sacs demonstrated the presence of active Ca absorption with serosal-to-mucosal (S/M) ratio of 2.65 +/- 0.20 (n = 6), which was greater than unity (P less than 0.001). In contrast, the S/M for Mg did not exceed unity. Mucosal duodenal Ca uptake exhibited a large saturable (Michaelis constant of 4.80 +/- 0.34 mM, maximal velocity of 4.71 +/- 0.13 nmol.min-1.mg-1) and a small nonsaturable component (0.12 +/- 0.01 nmol.min-1.mg-1) in 5-wk-old rats (120 g). In 72-wk-old rats (600 g) the diffusional component of Ca uptake became predominant, and the slope increased significantly to 0.32 +/- 0.01 (P less than 0.05). Duodenal Mg uptake was completely concentration dependent and exhibited no age-related changes.

Biochemical markers for assessment of niacin status in young men: urinary and blood levels of niacin metabolites.

Biochemical markers of niacin status were studied in healthy young men fed 6.1 to 32 niacin equivalents (NE) per day over an 11-wk period while residing in a metabolic unit. Methylated metabolites of niacin, N1-methylnicotinamide (NMN) and N1-methyl-2-pyridone-5-carboxamide (2-pyr), in urine and plasma were determined during periods of low (6.1 or 10.1 NE per day), adequate (19 NE per day = 1 RDA) and high (25 or 32 NE per day) niacin intakes and after small test doses of nicotinamide. Urine excretion of less than 1.2 mg/d of either NMN or 2-pyr was a reliable indicator of subjects receiving the lowest intake of 6.1 NE/d, but the NMN metabolite was a better marker of subjects ingesting 10.1 NE/d. The ratio of 2-pyr/NMN in urine was not as good a measure of the 6.1 NE/d intake as the individual metabolite excretions and was not responsive to the 10.1 NE/d intake. Plasma niacin metabolites were generally not as reliable as urinary metabolites for identifying subjects receiving low niacin intakes, however, values for plasma 2-pyr dropped quickly and were eventually nondetectable. After a 1 RDA oral dose of nicotinamide, increases in urine and plasma 2-pyr levels above pre-dose baseline values were significantly decreased in subjects receiving low, as compared to adequate, niacin intake. A leucine supplement had no effect on the rate of repletion of niacin-deficient subjects nor on the level of methylated niacin metabolites in urine or plasma.

Heat shock inhibits radiation-induced activation of NF-kappaB via inhibition of I-kappaB kinase.

Radiation stimulates signaling cascades that result in the activation of several transcription factors that are believed to play a central role in protective response(s) to ionizing radiation (IR). It is also well established that heat shock alters the regulation of signaling cascades and transcription factors and is a potent radiosensitizing agent. To explore the hypothesis that heat disrupts or alters the regulation of signaling factors activated by IR, the effect of heat shock on IR-induced activation of NF-kappaB was determined. Irradiated HeLa cells demonstrated transient increases in NF-kappaB DNA binding activity and NF-kappaB protein nuclear localization. In addition, irradiated cells demonstrated increased I-kappaB phosphorylation and decreased I-kappaBalpha cytoplasmic protein levels, corresponding temporally with the increase of NF-kappaB DNA binding. Heat shock prior to IR inhibited the increase in NF-kappaB DNA binding activity, nuclear localization of NF-kappaB, and the phosphorylation and subsequent degradation of I-kappaB. I-kappaB kinase (IKK) immunoprecipitation assays demonstrated an increase in IKK catalytic activity in response to IR that was inhibited by pretreatment with heat. Kinetic experiments determined that heat-induced inhibition of NF-kappaB activation in response to IR decayed within 5 h after heating. Furthermore, pretreatment with cycloheximide, to block de novo protein synthesis, did not alter heat shock inhibition of IR induction of NF-kappaB. These experiments demonstrate that heat shock transiently inhibits IR induction of NF-kappaB DNA binding activity by preventing IKK activation and suggests a mechanism independent of protein synthesis.