Antitumor effect of a copper coordination compound with superoxide dismutase-like activity.

Growth of Ehrlich carcinomas in inbred CBA mice was retarded by im administration of Cu(II)(3,5-diisopropylsalicylate)2 (CuDIPS). CuDIPS is a low molecular weight (mol wt = 503) copper coordination compound that exhibits superoxide dismutase (SOD)-like activity. It has been used as an anti-inflammatory agent and is lipid-soluble. This property enables the compound to penetrate membranes, thus becoming an intracellular O2- scavenger. In the tumor system studied, the amounts of both copper- and zinc-containing SOD (CuZnSOD) and manganese-containing SOD are reduced. Injection of Orgotein (CuZnSOD from bovine liver) had no significant effect on tumor growth and host survival. When CuDIPS was administered at various doses, reduction in tumor size, delay of metastasis, and a significant increase in survival of the hosts were observed.

Possible role of glutathione in the antitumor effect of a copper-containing synthetic superoxide dismutase in mice.

The effect of glutathione and a glutathione reductase inhibitor on the antitumor effect of Cu(II)(3,5-diisopropylsalicylate)2 (CuDIPS) was studied. CuDIPS is a low-molecular-weight copper coordination compound that exhibits superoxide dismutase-like activity. CuDIPS had antitumor activity against intraperitoneal Ehrlich ascites carcinoma in Swiss mice. A single ip injection of glutathione partially eliminated the antitumor effect of CuDIPS, whereas a single ip injection of 1,3-bis(2-chloroethyl)-1-nitrosourea enhanced the antitumor effect of CuDIPS. These results are consistent with the hypothesis that CuDIPS exerts part of its antitumor effect by producing H2O2.

Postirradiation treatment with copper(II)2(3,5-diisopropylsalicylate)4 enhances radiation recovery and hemopoietic regeneration.

We have previously reported that copper(II)2(3,5-diisopropylsalicylate)4 (Cu-DIPS), administered 3 h before exposure to lethal irradiation, significantly increased the survival rate of mice. Agents that can improve recovery from irradiation are of particular importance for accidental radiation exposure if they are effective when given after exposure. In the present study, we showed that Cu-DIPS had radiation recovery activity when administered subsequent to radiation exposure. Mice were exposed to 800 cGy irradiation and 3 h later injected with vehicle or 20, 40, or 60 mumol/kg Cu-DIPS. The 30-day survival rate was significantly increased at all doses of Cu-DIPS tested. Survival increased from 47% for vehicle-treated mice to 78% (p less than 0.001) for mice treated with 40 mumol/kg. The recovery of hemopoietic activity was assessed in similarly treated mice 14 and 24 days after irradiation. The postirradiation Cu-DIPS treatment significantly increased spleen weights, bone marrow cellularity, and hemopoietic activity in the spleen and bone marrow compared to vehicle-treated controls. Enhanced recovery of hemopoietic activity included both committed progenitor granulocyte-macrophage colony-forming units (GM-CFU) and more primitive stem cells (endogenous spleen colony-forming units, CFU-Se). The number of CFU-Se at 14 days, the number of bone marrow GM-CFU at 24 days, and bone marrow cellularity at 24 days appear to be better predictors of survival rates than other parameters.

Copper(II)2(3,5-diisopropylsalicylate)4 stimulates hemopoiesis in normal and irradiated mice.

We have previously reported that copper(II)2(3,5-diisopropylsalicylate)4 (Cu-DIPS) significantly increased the survival rate of mice exposed to lethal irradiation. To examine whether Cu-DIPS affected hemopoietic activity, groups of mice were treated with Cu-DIPS or vehicle and assayed for in vitro interleukin 3 (IL-3)-dependent colony-forming units (CFU-C) and for committed progenitor granulocyte-macrophage CFU (GM-CFU). Cu-DIPS increased the number of splenic IL-3 CFU-C by five- to sixfold 7 days after treatment and splenic GM-CFU by 12-fold on day 24. These increases were accompanied by a 50% increase in spleen weight. Bone marrow IL-3 CFU-C and GM-CFU were not affected at 7 or 14 days after treatment, but were somewhat depressed at 24 days. In irradiated (8.0 Gy) mice treated with Cu-DIPS or vehicle, splenic IL-3 CFU-C and GM-CFU were undetectable 7 days after irradiation, but recovered more rapidly in Cu-DIPS-treated mice. By 24 days splenic IL-3 CFU-C in Cu-DIPS-treated mice recovered to 150% of normal (unirradiated) values and GM-CFU recovered to 270% of normal, whereas irradiated control values remained at 25% and 7%, respectively. The recovery of bone marrow hemopoiesis was slower than spleen, but 42 days after irradiation Cu-DIPS-treated mice had higher levels of bone marrow IL-3 CFU-C (eightfold) and GM-CFU (4.6-fold) than vehicle-treated mice. Cu-DIPS stimulated sixfold increases in renewable, pluripotent stem cells as measured by the in vivo assay of endogenous colony-forming units (CFU-Se).

Lack of effects of copper gluconate supplementation.

A double-blind study was done giving 10 mg of copper/day as copper gluconate or placebo capsules for 12 wk. The seven subjects receiving copper gluconate had no change in the level of copper in the serum, urine, or hair. There was also no change in the levels of zinc or magnesium. There was also no significant change in levels of hematocrit, triglyceride, SGOT, GGT, LDH, cholesterol, or alkaline phosphatase. The side effects of nausea, diarrhea, and heartburn were the same in the subjects receiving copper gluconate and subjects receiving placebo capsules.

Bis(3,5-diisopropylsalicylato)copper(II), a potent radioprotectant with superoxide dismutase mimetic activity.

Superoxide disproportionation may partially account for the noteworthy radioprotectant activity of bis(3,5-diisopropylsalicylato)copper(II) [CuII(3,5-dips)2]. Groups of mice treated with CuII(3,5-dips)2 3 or 24 h before exposure to a lethal dose of gamma-radiation had survival rates of 33% and 58%, respectively. These results suggest that copper complexes might be developed for protection of normal tissues in association with cancer radiotherapy and protection against occupational exposures to hazardous radiation.

Commercial discharge packs and breast-feeding counseling: effects on infant-feeding practices in a randomized trial.

A randomized controlled trial was conducted to evaluate two interventions for prolonging the duration of breast-feeding in a multiethnic sample of 343 low-income urban women. One intervention compared research breast-feeding bedside counseling by a trained counselor, who also made eight telephone calls during the first 3 months of the infant's life, with the routine breast-feeding counseling provided in the hospital by nurses. The other intervention compared commercial discharge packs provided by formula companies with research discharge packs designed to be consistent with the WHO Code of Marketing of Breastmilk Substitutes. When infants were 4 months old, a telephone interviewer unaware of treatment status contacted 95% (324/343) of the women to determine the infants' feeding and health histories. Compared with routine counseling, research counseling delayed the first introduction of solid foods to the infant's diet (P = .03, one-tailed) but did not exert a statistically significant effect on breast-feeding by 4 months' postpartum. Women who received the research discharge pack, compared with those who received the commercial pack, were more likely to prolong exclusive breast-feeding (P = .004, one-tailed), to be partially breast-feeding at 4 months postpartum (P = .04, one-tailed), and to delay the daily use of solid foods in the infant's diet (P = .017, one-tailed). Among the women who received research counseling, the research discharge pack was associated with lower rates of rehospitalization of infants than was the commercial pack (1% v 14%; P = .014, two-tailed). We conclude that in high-risk maternity populations, commercial discharge materials for breast-feeding women should be replaced by materials consistent with the WHO Code.

Pediatrician involvement in breast-feeding promotion: a national study of residents and practitioners.

OBJECTIVE: Physician support for breast-feeding mothers has been shown to improve breast-feeding rates, but no evaluation of the adequacy of physicians' breast-feeding-related training has been conducted. This study was designed to assess pediatricians' knowledge, attitudes, training, and activities related to breast-feeding promotion. METHODS: Surveys were mailed to a national random sample of pediatric residents (n = 999) and practitioners (n = 610) who were board certified within the previous 3 to 5 years. RESULTS: Response rates were 74% for residents and 69% for practitioners. Although more than 90% of respondents agreed that pediatricians should be involved in breast-feeding promotion, their clinical knowledge and experience did not suggest a high degree of competency. For example, practitioners were only slightly more aware of breast-feeding's protective effect against otitis media (71% vs 60%), and more than one quarter of both groups did not agree that exclusive breast-feeding is the most beneficial form of infant nutrition. Clinical advice often included inappropriate recommendations for breast-feeding termination or formula supplementation; only 64% of practitioners and 52% of residents knew that supplementing during the first few weeks of life may cause breast-feeding failure. For both groups, prior personal breast-feeding experience (ie, respondent or spouse had breast-fed an infant for 2 or more weeks) was a major determinant of improved clinical knowledge, more frequent activity, and greater self-confidence and perceived effectiveness in the area of breast-feeding promotion. Residents reported that the breast-feeding instruction provided during training was primarily in lecture format, with limited clinical opportunities to practice skills needed to assist breast-feeding mothers. Reflecting on their own training, more than 70% of practitioners recommended that more time be devoted to direct patient interaction and practice of counseling and problem-solving skills. CONCLUSIONS: These results indicate that residency training does not adequately prepare pediatricians for their role in breast-feeding promotion. Improvements in residency training and innovative continuing education programs should be implemented to help pediatricians meet the needs of their breast-feeding patients.

Parental response to repeat testing of infants with 'false-positive' results in a newborn screening program.

Forty-five percent of the parents of 60 infants being retested in a newborn screening program for metabolic disorders understood that their infant was undergoing retesting because the first test result was abnormal. Fifty-five percent had incorrect or incomplete information, believing that retesting was routine, or that mistakes had been made in the original testing procedure, or they report being told nothing specific about the testing. Parents who were aware that the initial test was abnormal were no more anxious or depressed while waiting for the repeat test results than other parents. At a second interview after learning the normal results of the repeat test, both those parents informed of the initial abnormal result as well as those not informed were less anxious and depressed. However, 36% of the parents of these normal infants reported concern about the health of their infant because of the repeat testing. This concern was not related to a parent's knowledge that the initial test result was abnormal, but was greater in parents reporting that they had not received sufficient information about the screening/testing process and its significance for the health of their infant.

Stable superoxide dismutase (SOD)-mimetic ternary human serum albumin-Cu(II)(3,5-diisopropylsalicylate)2/Cu(II)2(3,5-diisopropylsalicylate)4 complexes in tissue distribution of the binary complex.

Copper(II)2(3,5-diisopropylsalicylate)4 [Cu(II)2(3,5-DIPS)4] has been found to have antiinflammatory, antiulcer, anticancer, anticonvulsant, antimutagenic, antidiabetic, analgesic, and radiation protection and recovery activities. It has also been found to reduce ischemia-reperfusion injury. Because of these activities it was of interest to understand how this compound is transported in the body to affected tissues. Evidence supporting the suggested formation of ternary human serum albumin (HSA)-Cu(II)(3,5-DIPS)2 or Cu(II)2(3,5-DIPS)4 complexes was obtained using ultraviolet spectrophotometry, dialysis, and atomic absorption spectrophotometry or atomic emission spectroscopy. Superoxide dismutase (SOD)-mimetic activity was also determined using the xanthine/xanthine oxidase/cytochrome c system. Ultraviolet spectra of aqueous solution mixtures of Cu(II)2(3,5-DIPS)4 in equilibrium with 2Cu(II)(3,5-DIPS)2 and HSA as well as aqueous solutions of solid Cu(II)2(3,5-DIPS)4 obtained by stirring the solid with an aqueous solution of HSA showed no obvious change in absorbance to indicate ternary complex formation. However, comparison of ultraviolet spectra taken before and after dialysis supports the suggested bonding of Cu(II)(3,5-DIPS)2 or Cu(II)2(3,5-DIPS)4 to HSA. Comparison of copper concentrations before and after dialysis also supports the suggested bonding of Cu(II)(3,5-DIPS)2 or Cu(II)2(3,5-DIPS)4 to HSA. Based upon these data it is plausible that Cu(II)(3,5-DIPS)2 or Cu(II)2(3,5-DIPS)4 form stable ternary complexes with HSA. These stable ternary complexes were also found to have SOD-mimetic activity.

Non-steroidal anti-inflammatory drug-copper complex modulation of polymorphonuclear leukocyte migration.

These studies were intended to compare the effects of aspirin, 3,5-diisopropysalicylic acid (3,5-DIPS), and indomethacin with those of their copper complexes: Cu(II)2(aspirinate)4, Cu(II)2(3,5-DIPS)4, and Cu(II)2(indomethacinate)4 as well as Cu(II)2(acetate)4 on polymorphonuclear leukocyte (PMNL) random and directional migration, in addition to their anti-inflammatory activities. Experiments were performed both in vivo and in vitro. In vitro modifications of PMNL migration were measured with the Boyden chamber using N-formyl-methionyl-leucyl-phenylalanine (fMLP) as the chemoattractant and in the agarose assay using fMLP and serum chemotactic derivatives of complement as chemoattractants. In vivo anti-inflammatory activities of these compounds were determined after induction of a serum-induced pleurisy in the rat, and measurement of exudate volume and number of exudative cells 4 hr later. Copper complexes of non-steroidal anti-inflammatory drugs (NSAIDs) were found to be more effective in decreasing random migration and chemotaxis of PMNLs than their parent drugs or Cu(II)2(acetate)4 in in vitro studies. Only chemotaxis was found to be reduced significantly for PMNLs obtained from pleuritic rats after in vivo treatment and the order of copper complex effectiveness was: Cu(II)2(indomethacinate)4 greater than Cu(II)2(3,5-DIPS)4 greater than Cu(II)2(aspirinate)4. All doses of Cu(II)2(acetate)4 administered in vivo failed to affect chemotactic activity. Copper complexes of NSAIDs were also more effective than their parent drugs as anti-inflammatory agents, and Cu(II)2(acetate)4 had no anti-inflammatory activity in this model of inflammation. The order of anti-inflammatory activity was: Cu(II)2(indomethacinate)4 greater than Cu(II)2(3,5-DIPS)4 greater than Cu(II)2(aspirinate)4.

Aluminum in the environment and human health.

The review of over 800 references on aluminum (Al) published since the mid-fifties covers the occurrence of Al in soil, air, water, plants and food products, as well as air and water pollution problems. In addition, the existing quality criteria, the biology and toxicology of Al, and the therapeutic and medical uses are presented. It is concluded that absorption and retention or accumulation of Al in humans occurs at lower levels of intake than had been assumed formerly. However, levels of 5 to 50 times the normal daily intake do not appear to interfere with other metabolic processes. The adverse effects of Al reported in the more recent years resulted from the inhalation or ingestion of Al in concentrations many times greater than the amounts present under normal circumstances. As in the past, there is still no need for concern by the public or producers of Al or its products concerning hazards to human health derived from well established and extensively used products.

Couple agreement before and after genetic counseling.

Before receiving genetic counseling, 699 couples completed questionnaires that were identical for both spouses. Of 385 couples where both spouses indicated a major reason for seeking counseling, 45% identified the same major reason while 55% identified different reasons. In 74% of 542 fertile couples, both spouses had identical short-term reproductive plans: 60% desired the same ideal number of children; and 44% perceived the same level of risk of having an affected child. Agreement on the seriousness of eleven potential problems occasioned by an affected child ranged from 55% to 67%. Couples were asked to return questionnaires within 7 to 10 days after counseling. At this time 76% of spouses agreed about short-term reproductive plans; 66% agreed about longer-term reproductive plans; and 60% agreed about ideal number of children. There were statistically significant increases in agreement about risk interpretation and about six of eleven potential problems in raising an affected child. The data suggest that a substantial number of couples come to genetic counseling with varying concerns and reproductive plans. There is, in general, as much disagreement on these issues between spouses after as before counseling. Implications for counseling are discussed.

Genetic counseling and reproductive uncertainty.

Of 836 fertile women seeking genetic counseling, 544 (65%) reported that their major reason for doing so was to obtain information to help in deciding if they should have a child. Thirty-four percent of these 836 women entered counseling uncertain about undertaking a pregnancy in the next 2 years. After counseling, 28% of the 836 had uncertain pregnancy intentions. These included 66% of those who were uncertain and 11% of those who were reproductively certain before counseling. Stepwise logistic regression identified the following as independently and significantly associated with reproductive uncertainty after counseling: 1) uncertainty before counseling; 2) uncertainty about ideal family size; 3) concern about the effects of an affected child on the client's social life; 4) perceived serious problems caring for a child with a birth defect now living at home; and 5) new concerns raised in counseling. Reproductive uncertainty after counseling was not related to characteristics of the risked birth defect, level of risk, treatment potential, or client learning of medical and genetic facts. These data suggest that genetic counseling will not eliminate reproductive uncertainty for many clients, because this uncertainty is related to factors mainly outside the usual scope of counseling.

Reproductive plans of genetic counseling clients not eligible for prenatal diagnosis.

A prospective study of the reproductive plans of 185 genetic counseling clients at risk for birth defects not diagnosable prenatally found, 6 months after counseling, 1) a small increase in the number of pregnancies initiated and planned, compared to pregnancies planned before counseling; 2) an increase in initiated and planned pregnancies among clients at both high as well as low risk; and 3) reproductive plans after counseling more closely correlated with clients' perceptions of the social, familial, and economic burdens of an affected child than with medically defined risk and specific clinical characteristics of the birth defects. In discussing the burden of a birth defect with clients, counselors are encouraged to discuss not only the medical burden, but the social, familial, and financial burdens as well.

Acceptance of home and clinic-based cystic fibrosis carrier education and testing by first, second, and third degree relatives of cystic fibrosis patients.

We contacted and offered free cystic fibrosis (CF) carrier education and testing to the first, second, and third degree relatives of individuals with CF followed at a large Southeastern US CF Clinic. Relatives were offered CF carrier education and testing either in their homes or in a genetic counseling clinic. Overall, of 514 relatives offered free CF carrier education and testing, 299 (58%) accepted. Significantly more (67%) of those offered education and testing in their homes accepted than those offered education and testing in a genetic counseling clinic (45%). Regression analyses identified several factors, including education, income, gender, perceived chance of being a carrier, and perceived chance of having a child who is a CF carrier, as predictors of acceptance of education and testing in both home and clinic sites. A smaller set of factors was identified that predicted acceptance of education and testing unique to each site. Within the limits of this study and its design, even when CF carrier testing is offered free of charge, including education and testing in the home, acceptance of education and testing, while higher than in general population samples, is not universal among at-risk relatives. Several factors which may have contributed to the observations reported in this study are discussed.

Proband and parent assistance in identifying relatives for cystic fibrosis carrier testing.

To identify, contact, and offer free cystic fibrosis (CF) carrier education, testing, and genetic counseling to the first, second, and third degree relatives of individuals with CF, study personnel contacted probands or the parents of minor probands requesting assistance in identifying relatives. We requested family pedigrees, including names, addresses, and phone numbers and if necessary a saliva sample for determination of the specific CF mutations in the family. Two hundred three families of 220 probands being followed at a large CF clinic in the Southeastern United States were eligible for inclusion in the study. Of the 203 families 109 (53.7%) assisted by providing contact information on relatives and, when necessary, a saliva sample for mutation analysis. An additional 33 (16.4%) agreed to assist but did not provide either or both contact information or saliva samples. Sixty-one (30.1%) declined to provide assistance. Thirteen percent of the probands/parents wanted to talk with relatives before providing contact information. A logistic regression model predicting proband/parent assistance is provided. This study suggests that the active outreach method used here to identify at risk relatives to offer them CF carrier testing resulted in somewhat lower proband or parent assistance than reported by other similar approaches. The strengths and weaknesses of this approach, including comments by probands and parents on the method, are discussed.

Essential metalloelement metabolism and radiation protection and recovery.

Understanding the metabolism of essential metalloelements and their role in tissue maintenance and function as well as the roles of essential metalloelement-dependent enzymes in responding to injury offers a new approach to preventing and/or treating radiation injury. This review presents the roles of some essential metalloelement-dependent enzymes in the maintenance and function of tissues and their responses to radiation injury and gives an account of the observed effects of nontoxic doses of essential metalloelement compounds on protection against radiation damage and its recovery. The radiolysis of chemical bonds and free radicals derived from oxygen accounts for the acute and chronic aspects of radiation injury. The recognized biochemical roles of essential metalloelements and their observed pharmacological effects predict the therapeutic usefulness of essential metalloelement complexes in the prevention and/or treatment of radiation injury. Copper complexes have radiation protection and radiation recovery activities and cause rapid recovery of immunocompetence and radiation-induced damage to cells and tissues. Recently, iron, manganese, and zinc complexes have also been found to prevent death in lethally irradiated mice. These pharmacological effects of essential metalloelement complexes can be understood to be due to facilitation of de novo synthesis of essential metalloelement-dependent enzymes which have roles in preventing the accumulation of pathological concentrations of oxygen radicals or repairing damage caused by radiation-induced bond homolysis. Essential metalloelement complexes offer a physiological approach to prevention and/or treatment of radiation injury.

Radiorecovery activity of manganese(III)2(II)(mu 3-O)-(mu-3,5-diisopropylsalicylate)6.

Manganese(III)2(II)(mu 3-O)(mu-3,5-diisopropylsalicylate)6 [Mn3(O)(3,5-DIPS)6] was used to treat female C57BL/6 mice irradiated with LD50/30 doses of gamma rays and examine the possibility that treatment after irradiation increases survival. Female C57BL/6 mice were treated with 0, 10, 20, or 40 mumol Mn3(O)(3,5-DIPS)6/kg of body mass 1 or 3 h after irradiation. Treatment with 40 mumol/kg 1 or 3 h after irradiation produced survivals of 72 or 92%, respectively, increases of 29 or 130% in comparison with 56 or 40% survivals in the respective vehicle-treated groups. These data support the hypothesis that Mn3(O)(3,5-DIPS)6 is an effective radiorecovery agent.

Evaluation of the anticancer activities of Tweens 20, 40 and 60 in SJL/J mice.

Tween 20 (polyoxyethylenesorbitan monolaurate), Tween 40 (polyoxyethylenesorbitan monopalmitate), and Tween 60 (polyoxyethylenesorbitan monostearate) were found to have little activity against development of reticulum cell sarcoma in SJL/J mice. These findings resulted in studies to explain the previous observation that Tween 80 (polyoxyethylenesorbitan monooleate) has anticancer activity in the SJL/J tumor system. Evidence is presented to suggest that the apparent difference may be related to the fatty acid component (saturated or unsaturated) of the Tween.