Protective effect of Allium sativum (garlic) aqueous extract against lead-induced oxidative stress in the rat brain, liver, and kidney.

The present investigation was undertaken to evaluate the ameliorative activity of Allium sativum against lead-induced oxidative stress in the brain, liver, and kidney of male rats. Four groups of male Wistar strain rats (100-120 g) were taken: group 1 received 1000 mg/L sodium acetate and group 2 was given 1000 mg/L lead acetate through drinking water for 2 weeks. Group 3 and 4 were treated with 250 mg/kg body weight/day of A. sativum and 500 mg/kg body weight/day of A. sativum, respectively, by oral intubation for a period of 2 weeks along with lead acetate. The rats were sacrificed after treatment and the brain, liver, and kidney were isolated on ice. In the brain, four important regions namely the hippocampus, cerebellum, cerebral cortex, and brain stem were separated and used for the present investigation. Blood was also drawn by cardiac puncture and preserved in heparinized vials at 4 °C for estimation of delta-aminolevulinic acid dehydratase (ALAD) activity. The results showed a significant (p < 0.05) increase in reactive oxygen species (ROS), lipid peroxidation products (LPP), total protein carbonyl content (TPCC), and lead in the selected brain regions, liver, and kidney of lead-exposed group compared with their respective controls. Blood delta-ALAD activity showed a significant (p < 0.05) decrease in the lead-exposed rats. However, the concomitant administration of A. sativum resulted in tissue-specific recovery of oxidative stress parameters namely ROS, LPP, and TPCC. A. sativum treatment also restored the blood delta-ALAD activity back to control. Overall, our results indicate that A. sativum administration could be an effective antioxidant treatment strategy for lead-induced oxidative insult.

Erectile function recovery in patients after non-nerve sparing radical prostatectomy.

Few studies have looked at erectile function recovery (EFR) rates in men undergoing non-nerve sparing resection during radical prostatectomy (RP). Existing studies show great variation in EFR rates owing to multiple factors that minimize their utility in counselling RP patients. We investigated the EFR rate and its predictors in unilateral cavernous nerve resection and bilateral cavernous nerve resection patients 24 months after RP. We conducted a population-based, prospective cohort study of 966 patients who underwent RP at a tertiary cancer centre from 2008 to 2012. Cavernous nerve condition was evaluated on a 4-point nerve sparing score and assigned to one of three groups: bilateral sparing, unilateral resection (UNR) and bilateral nerve resection (BNR). EF was assessed pre-RP and 24-30 months post-op using a validated 5-point patient-reported scale (1 = fully rigid; 5 = no tumescence). EFR was defined as a post-op EF grade of 1-2. Statistical analysis included descriptive statistics, anova, chi-square, Fisher's exact test and logistic regression. Mean baseline EF was 1.84 ± 1.3 and 2.74 ± 1.5 for UNR and BNR patients respectively. Thirty-three percent of UNR patients and 13% of BNR patients exhibited EFR. Age, baseline EF were predictors of EFR. Multivariable analysis showed baseline EF was a significant predictor of EFR at 24 months for UNR. For BNR patients, pre-RP EF was the only factor predictive of EFR. Patients undergoing nerve resection still have a significant chance of achieving true EFR, with UNR surgery patients showing more potential for improvement than patients undergoing BNR surgery. Age and baseline EFR characterize recovery prospects in these two groups. Physicians should thus measure and account for baseline EF in addition to age and the degree of nerve resection when advising patients about expectations for successful EF following RP.

Involvement of substance P and central opioid receptors in morphine modulation of the CHS response.

Morphine administration prior to challenge with the antigen 2,4-dinitro-fluorobenzene increases the contact hypersensitivity (CHS) response in rats. The present study extended these findings by showing that central, but not systemic, administration of N-methylnaltrexone antagonized the morphine-induced enhancement of the CHS response. The importance of the neuroimmune mediator substance P was shown via the attenuation of the morphine-induced enhancement following both systemic and topical administration of the NK-1 antagonist WIN51,708. Taken together, the findings of the present study provide new data showing that central opioid receptors and peripheral substance P are involved in the morphine-induced enhancement of the CHS response.

Morphine modulation of the contact hypersensitivity response: characterization of immunological changes.

Previous investigations in our laboratory showed that systemic morphine administration 1 h prior to elicitation of the in vivo contact hypersensitivity (CHS) response produced a robust increase in inflammation at the site of antigen reexposure. The present study extended those findings by characterizing the effect of morphine on immunological processes important in the development of the CHS response. To induce contact hypersensitivity, the antigen 2,4-dinitrofluorobenzene was applied to the pinnae of previously sensitized rats. Morphine administration produced an increase in inducible nitric oxide synthase mRNA and the proinflammatory cytokine interleukin-6, at the site of antigen reexposure. In contrast, morphine did not alter expression of the anti-inflammatory cytokine interleukin-10. Morphine also produced an increase in the proliferation of lymphocytes from the peripheral (i.e., cervical) lymph nodes when assessed 72 h following challenge. These studies show that the morphine-induced increase in the in vivo CHS response involves immunologically specific alterations.

Naltrexone administration attenuates surgery-induced immune alterations in rats.

Surgery is a commonly performed procedure which produces substantial alterations in immune function in both humans and animals. To better understand the mechanism of surgery-induced immunomodulation, the present study investigated the effect of the opioid antagonist naltrexone on surgery-induced immune alterations in rats. Based on previous investigations in our laboratory, rats underwent a 6-cm laparotomy with no internal manipulation and immunological assessments were completed 24 h following the surgical procedure. Naltrexone was administered at the time of surgery and every 4 h thereafter until immune assessment. Results showed that naltrexone attenuated the surgery-induced decrease in natural killer cell cytotoxicity, B-cell proliferation, T-cell proliferation, and production of the cytokine IFN-gamma. These results are among the first to show that pharmacological antagonism of opioid receptors can prevent deleterious immune changes in the postoperative state, suggesting a detrimental role of the endogenous opioids in surgical procedures.

pH dependence of formation of a partially unfolded state of a Lys 73 --> His variant of iso-1-cytochrome c: implications for the alkaline conformational transition of cytochrome c.

The alkaline conformational transition of a lysine 73 --> histidine variant of iso-1-cytochrome c has been studied. The transition has been monitored at 695 nm, a band sensitive to the presence of the heme-methionine 80 bond, at the heme Soret band which is sensitive to the nature of the heme ligand, and by NMR methods. The guanidine hydrochloride dependence of the alkaline conformational transition has also been monitored. The histidine 73 protein has an unusual biphasic alkaline conformational transition at both 695 nm and the heme Soret band, consistent with a three-state process. The conformational transition is fully reversible. An equilibrium model has been developed to account for this behavior. With this model, it has been possible to obtain the acid constant for the trigger group, pK(H), of the low-pH phase from the equilibrium data. A pK(H) value of 6.6 +/- 0.1 in H(2)O was obtained, consistent with a histidine acting as the trigger group. The NMR data for the low-pH phase of the alkaline conformational transition are consistent with an imidazole ligand replacing Met 80. For the high-pH phase of the biphasic alkaline transition, the NMR data are consistent with lysine 79 being the heme ligand. Guanidine hydrochloride m values of 1.67 +/- 0.08 and 1.1 +/- 0.2 kcal mol(-1) M(-1) were obtained for the low- and high-pH phases of the biphasic alkaline transition of the histidine 73 protein, respectively, consistent with a greater structural disruption for the low-pH phase of the transition.

Cloning and functional analysis of sucrose:sucrose 1-fructosyltransferase from tall fescue.

Enzymes of grasses involved in fructan synthesis are of interest since they play a major role in assimilate partitioning and allocation, for instance in the leaf growth zone. Several fructosyltransferases from tall fescue (Festuca arundinacea) have previously been purified (Lüscher and Nelson, 1995). It is surprising that all of these enzyme preparations appeared to act both as sucrose (Suc):Suc 1-fructosyl transferases (1-SST) and as fructan:fructan 6(G)-fructosyl transferases. Here we report the cloning of a cDNA corresponding to the predominant protein in one of the fructosyl transferase preparations, its transient expression in tobacco protoplasts, and its functional analysis in the methylotrophic yeast, Pichia pastoris. When the cDNA was transiently expressed in tobacco protoplasts, the corresponding enzyme preparations produced 1-kestose from Suc, showing that the cDNA encodes a 1-SST. When the cDNA was expressed in P. pastoris, the recombinant protein had all the properties of known 1-SSTs, namely 1-kestose production, moderate nystose production, lack of 6-kestose production, and fructan exohydrolase activity with 1-kestose as the substrate. The physical properties were similar to those of the previously purified enzyme, except for its apparent lack of fructan:fructan 6(G)-fructosyl transferase activity. The expression pattern of the corresponding mRNA was studied in different zones of the growing leaves, and it was shown that transcript levels matched the 1-SST activity and fructan content.

Endomorphin-1 induces antinociception without immunomodulatory effects in the rat.

RATIONALE: Although there is evidence that central opioid receptors are involved in immunomodulation, it has been only recently that an endogenous agonist, designated endomorphin-1, possessing high selectivity and affinity for the mu opioid receptor has been identified. OBJECTIVE: The present study assesses the immunomodulatory effects of endomorphin- in the rat and provides further evaluation of the antinociceptive effects of endomorphin-1. METHODS: Rats were surgically implanted with cannulae directed at the lateral cerebral ventricle. Animals received vehicle or endomorphin-1 at doses of 31.63 or 56.23 microg (ICV) and were tested for antinociception in two different assays, the warm water tail withdrawal procedure and the hotplate assay. Additional studies assessed the effect of naltrexone on the antinociception produced by endomorphin-1 in both antinociceptive assessments. Assessments of immune status following endomorphin-1 treatment included measurements of splenic natural killer cell activity, production of interferon-y, and lymphocyte proliferative responses to mitogenic stimulation by Con-A, LPS, and the microbial superantigen, TSST-1. RESULTS: Endomorphin-1 induced significant and naltrexone reversible antinociception 30 and 60 min following drug administration, as measured by the hotplate assay and warm water tail withdrawal procedure. In marked contrast, endomorphin-1 did not produce immunomodulatory effects up to 120 min following ICV administration. CONCLUSIONS: Endomorphin-1 produces antinociception but does not induce immunomodulatory effects in the rat. These findings suggest that it is possible to develop therapeutic strategies for separating antinociception and immunomodulatory properties through the mu opioid receptor.

Involvement of central mu- but not delta- or kappa-opioid receptors in immunomodulation.

Studies completed in both humans and animals have shown that opioids have significant effects on the immune system via pharmacological interactions with the opioid receptor. However, the type of opioid receptor at which morphine binding produces changes in immune status has not been well characterized. To determine the type of opioid receptor involved in opioid-induced immune alterations, the present study assessed the effects of agonists selective for the mu-, delta-, and kappa-opioid receptors. The site of action (i.e., peripheral vs central) at which opioids produce immune changes was investigated by injecting the agonists directly into the left lateral ventricle of the brain. Specifically, Lewis rats received an intracerebroventricular administration of [d-Ala(2),N-Me-Phe(4), Gly-ol(5)]enkephalin (DAMGO), a mu-receptor selective agonist, [D-Pen(2,5)]enkephalin (DPDPE), a delta-opioid receptor agonist, or U69,593, a kappa-receptor agonist. Immune assessments completed 1 h following drug administration showed that the mu-receptor selective agonist DAMGO produced a dose-dependent decrease in natural killer cell activity and T-lymphocyte proliferation to the mitogen concanavalin A (Con A); no immunological changes were found following DPDPE or U69,593 treatment. Calculation of the number of white blood cells per sample showed no differences between rats treated with saline and rats treated with any of the selective agonists. Administration of the opioid antagonist N-methylnaltrexone prior to DAMGO treatment attenuated the DAMGO-induced changes in immune status. Results from the present study indicate that the immunomodulatory effects of opioids can be attributed to interactions with the mu-opioid receptor.

The schedule of attitudes toward hastened death: Measuring desire for death in terminally ill cancer patients.

BACKGROUND: The authors examined the reliability and validity of the Schedule of Attitudes toward Hastened Death (SAHD), a self-report measure of desire for death previously validated in a population of individuals with the acquired immunodeficiency syndrome (AIDS), among terminally ill patients with cancer. METHODS: The authors interviewed 92 terminally ill cancer patients, all with a life expectancy of < 6 months, after admission to a palliative care hospital. Patients were administered the SAHD, a clinician-rated measure of desire for death (the Desire for Death Rating Scale [DDRS]), and several measures of physical and psychosocial well-being. RESULTS: The average number of SAHD items endorsed was 4. 76 (standard deviation, 4.3); 15 patients (16.3%) endorsed > or = 10 items, indicating a high desire for death. Internal consistency was strong (coefficient alpha = 0.88, median item-total correlation = 0. 49), as were indices of convergent validity. Total SAHD scores were correlated significantly (correlation coefficient [r] = 0.67) with the DDRS, and somewhat less so with measures of depression (r = 0. 49) and hopelessness (r = 0.55). Lower, but substantial, correlations were observed between the SAHD and measures of spiritual well-being (r = -0.42), quality of life (r = -0.36), physical symptoms (r = 0.38), and symptom distress (r = 0.38). No significant correlation was observed between SAHD scores and social support (r = -0.06) or pain intensity (r = 0.16); however, pain-related functional interference and overall physical functioning were correlated significantly with SAHD scores (r = 0.31 and r = -0.23, respectively). CONCLUSIONS: The SAHD appears to be a reliable and valid measure of desire for death among terminally ill cancer patients. Coupled with previous research in patients with AIDS, these results support the utility of the SAHD for research addressing interest in hastened death in patients with a life-threatening medical illness.

Topics of conflict between parents and young adolescents.

ISSUES AND PURPOSE: Parents often examine and question interactions with their young teen and may ask the advice of healthcare professionals. Topics, frequency, and intensity of conflicts between young adolescents and parents were therefore examined. DESIGN AND METHODS: A descriptive survey using the 44-item Issues Checklist (Robin, 1975) with 163 parent and young adolescent (ages 11-14) dyads. RESULTS: Parents and teens were congruent about their reports of the topics, frequency, and intensity of conflict. Discussion of the topics generally was not angry. Mothers reported the greatest quantity of issues. Potentially sensitive topics such as substance use, dating, and sex were rarely approached by either parent or young adolescent. Sociodemographic characteristics did not distinguish or were not associated with IC scores. PRACTICE IMPLICATIONS: Conflict is a common component of the parent-young adolescent relationship. Families with children entering adolescence can expect conflict about issues that recur but usually are not that "hot". Anticipating topics may put conflict in perspective. Nurses help families resolve conflicts associated with day-to-day conflicts as a first step toward opening up larger, potentially sensitive topics.

Phenotypic and functional assessments of immune status in the rat spleen following acute heroin treatment.

Heroin use is associated with an increased incidence of several types of infections, including HIV. Yet few studies have assessed whether heroin produces pharmacological alterations of immune status that might contribute to the increased rate of infections amongst heroin users. The present study investigated whether a single administration of heroin to rats produces dose-dependent alterations in functional measures of immune status and in the distribution of leukocyte subsets in the spleen. The results showed that heroin produces a dose-dependent, naltrexone-reversible suppression of the concanavalin A-stimulated proliferation of T cells, lipopolysaccharide-stimulated proliferation of B cells, production of interferon-gamma and cytotoxicity of natural killer (NK) cells in the spleen. Heroin's suppressive effect on NK cell activity results in part from a heroin-induced decrease in the relative number of NKR-P1A(hi) CD3- NK cells in the spleen. Heroin also decreases the percent of a splenic granulocyte subset, the CD11b/c+ HIS48(hi) cells, whose function currently is unknown. In contrast, heroin does not alter relative numbers of CD4+ CD3+ T cells, CD8+ CD3+ T cells, CD45+ B cells, NKR-P1A(lo) CD3+ T cells, CD11b/c+ ED1+ (or CD11b/c+ HIS48-) monocytes/macrophages or CD11b/c+ ED1- (or CD11b/c+ HIS48+) total granulocytes in the spleen. Collectively, these findings demonstrate that heroin produces pharmacological effects on functional and phenotypic measures of immune status.

Depression, hopelessness, and desire for hastened death in terminally ill patients with cancer.

CONTEXT: Understanding why some terminally ill patients desire a hastened death has become an important issue in palliative care and the debate regarding legalization of assisted suicide. OBJECTIVES: To assess the prevalence of desire for hastened death among terminally ill cancer patients and to identify factors corresponding to desire for hastened death. Design Prospective survey conducted in a 200-bed palliative care hospital in New York, NY. PATIENTS: Ninety-two terminally ill cancer patients (60% female; 70% white; mean age, 65.9 years) admitted between June 1998 and January 1999 for end-of-life care who passed a cognitive screening test and provided sufficient data to permit analysis. MAIN OUTCOME MEASURE: Scores on the Schedule of Attitudes Toward Hastened Death (SAHD), a self-report measure assessing desire for hastened death among individuals with life-threatening medical illness. RESULTS: Sixteen patients (17%) were classified as having a high desire for hastened death based on the SAHD and 15 (16%) of 89 patients met criteria for a current major depressive episode. Desire for hastened death was significantly associated with a clinical diagnosis of depression (P=.001) as well as with measures of depressive symptom severity (P<.001) and hopelessness (P<.001). In multivariate analyses, depression (P=.003) and hopelessness (P<.001) provided independent and unique contributions to the prediction of desire for hastened death, while social support (P=.05) and physical functioning (P=.02) added significant but smaller contributions. CONCLUSIONS: Desire for hastened death among terminally ill cancer patients is not uncommon. Depression and hopelessness are the strongest predictors of desire for hastened death in this population and provide independent and unique contributions. Interventions addressing depression, hopelessness, and social support appear to be important aspects of adequate palliative care, particularly as it relates to desire for hastened death.

Enhancement of the contact hypersensitivity reaction by acute morphine administration at the elicitation phase.

The present study investigated the effects of morphine on the irritant contact sensitivity (ICS) and contact hypersensitivity (CHS) reaction. ICS was induced by croton oil application on the pinnae of naïve rats. Morphine injected prior to croton oil application did not affect the ICS response when assessed by measurements of pinnae thickness. CHS was induced by applying the antigen 2,4-dinitro-1-fluorobenzene (DNFB) to the pinnae of rats sensitized to DNFB. Rats received an injection of morphine prior to either initial antigen exposure (sensitization) or antigen reexposure (challenge). Morphine prior to challenge, but not sensitization, resulted in a pronounced enhancement of the CHS response as measured by pinna thickness. Quantitative PCR also showed increased IFN-gamma mRNA levels in the inflamed tissue of morphine-treated rats. Naltrexone blocked the morphine-induced enhancement of the CHS response. The differential effects of morphine suggest that opioids have a more pronounced effect on in vivo immune responses that involve immunological memory.

Managing nutrients across regions of the United States.

Nutrient balance in the ecosystem involves profitability of the agricultural enterprise and commitments to resource management to maintain quality of air, water, and land resources. Phosphorus and N are the two nutrients of major concern, and they behave differently in soils. Most P adheres strongly to soil particles and moves laterally with the soil during erosion processes, but with high concentrations more P remains in soluble forms and moves in the water fraction. Most N is soluble and moves laterally or downward with soil water. Soil scientists and agronomists have researched soil processes, plant nutrition, cropping systems, and water quality issues mainly on a field and farm level, but now the movement is to management and regulation of nonpoint problems on a watershed basis as proposed in the Clean Water Action Plan. The plan recognizes the vast diversity of soil parent materials and climates among geographic areas, even among and within watersheds, that determine crop adaptation and cropping systems, the role of states in regulatory processes, and the need for local citizens to have operational involvement. This process insures that nutrient management guidelines will be more site-specific and solutions can be focused on the direct problem. Directed efforts will be needed to educate local citizens, landowners, and caretakers of agricultural enterprises, and regulatory agencies. Several factors, including economic and social incentives for implementation must be considered along with the technologies available. The solutions are multidisciplinary, will require long-term research to accommodate climate variation, and should be associated with a strong commitment to education. Public funding will be needed to support the effort.

Dose-dependent effects of multiple acute cocaine injections on maternal behavior and aggression in Sprague-Dawley rats.

Rat dams, which had no prior drug treatment, were either nontreated controls or were injected subcutaneously 4 times during a 10-day period with a single dose of 30, 15 or 7.5 mg/kg of cocaine hydrochloride HCl, or normal saline. Injections were given immediately postpartum following delivery of their final pup (PPD 1), and again on postpartum day 3 (PPD 3), postpartum day 6 (PPD 6) and postpartum day 10 (PPD 10). Dams were observed 30 min following injections for maternal behavior (MB) towards 8 surrogate male pups on PPD 1 and PPD 3 and for aggression towards a male or female intruder in the presence of their litter on PPD 6 and PPD 10. Compared to saline and untreated controls, cocaine-treated dams exhibited more disruptions in MB on both PPD 1 and PPD 3 and were less aggressive towards an intruder, regardless of intruder sex, on PPD 6 and PPD 10. In most cases MB was altered in a dose-dependent manner with the higher doses of cocaine resulting in a greater disruption of behavior.

A dose-response study of chronic cocaine on maternal behavior in rats.

To determine if there was a dose-response relationship with regard to cocaine treatment and maternal behavior exhibited by lactating rats at doses that had not been previously investigated, we examined the effects of three doses of chronic cocaine administration throughout gestation on both onset and established maternal behavior. Dams were injected (SC) with 6.3, 13, or 25 mg/kg cocaine HCl or an equivalent volume of saline throughout gestation; maternal behavior was tested on postpartum days 1 and 3. At the doses employed, cocaine disrupted the onset of only one pup-directed component of maternal behavior significantly in a dose-response manner, although there were several statistically nonsignificant dose-dependent trends of behavioral disruptions. No pup-directed behaviors were disrupted during testing for established maternal behavior. These results indicate that gestational cocaine treatment at doses of 25 mg/kg and less have only minimal effects on the onset and no effect on the maintenance of maternal behavior using our paradigm. The relationship of the present findings to previous work is discussed.