Haloperidol-associated stealth liposomes: a potent carrier for delivering genes to human breast cancer cells

Sigma receptors are membrane-bound proteins that are overexpressed in certain human malignancies including breast cancer. These receptors show very high affinity for various sigma ligands including neuroleptics like haloperidol. We hypothesized that in associating haloperidol-linked lipid into the cationic lipid-DNA complex, we can specifically target and deliver genes to breast cancer cells that overexpress sigma receptors. In the present study, haloperidol was chemically modified to conjugate at the distal end of the polyethylene glycollinked phospholipid, which was then incorporated into the cationic liposome known to condense and deliver genes inside cells. The resulting haloperidol-conjugated targeted lipoplex showed at least 10-fold higher (p < 0.001) reporter gene expression in MCF-7 cells than control lipoplex. The reporter gene expression of the targeted lipoplex was significantly blocked by haloperidol (p < 0.001) and by another sigma ligand, 1,3-ditolylguanidine (p < 0.001) in the majority of cationic lipid to DNA charge ratios (+/-). Spironolactone-mediated sigma receptor down-regulation enabled MCF-7 to show 10-fold lower transgene expression with targeted lipoplex compared with that obtained in spironolactone-untreated cells. The targeted lipoplex generated nonspecific gene expression in sigma receptor-nonexpressing human cancer cells such as Hela, KB, HepG2, and Chinese hamster ovary cells. Moreover, the transgene expression remained unabated in physiologically relevant serum concentrations. This is the first study to demonstrate that haloperidol-targeted gene delivery systems can mediate efficient targeting of genes to sigma receptor-overexpressing breast cancer cells, thereby becoming a novel class of therapeutics for the treatment of human cancers.     

Effects of ethanol and haloperidol on plasma levels of hepatic enzymes, lipid profile, and apolipoprotein in rats.

This work studied the effects of ethanol in the absence and presence of haloperidol under two experimental conditions. In protocol 1, rats were treated daily with ethanol (4 g/kg, p.o.) for 7 days, and received only haloperidol (1 mg/kg, i.p.) from the 8th day to the 14th day. In protocol 2, animals received ethanol, and the treatment continued with ethanol and haloperidol from the 8th day to the 14th day. Results show increases in alanine transaminase (ALT; 48% and 55%) and aspartate transaminase (AST; 32% and 22%) levels after ethanol or haloperidol (14 days) treatments, as compared with controls. Apolipoprotein A-1 (APO A1) levels were increased by haloperidol, after 7- (148%) but not after 14-day treatments, as compared with controls. Levels of lipoprotein (high-density lipoprotein (HDL-C)) tended to be increased only by ethanol treatment for 14 days. ALT (80%) and AST (43%) levels were increased in the haloperidol plus ethanol group (protocol 2), as compared with controls. However, an increase in APO A1 levels was observed in the haloperidol group pretreated with ethanol (protocol 1), as compared with controls and ethanol 7-day treatments. Triglyceride (TG) levels were increased in the combination of ethanol and haloperidol in protocol 1 (234%) and 2 (106%), as compared with controls. Except for a small decrease in haloperidol groups, with or without ethanol, as related to ethanol alone, no other effect was observed in HDL-C levels. In conclusion, we showed that haloperidol might be effective in moderating lipid alterations caused by chronic alcohol intake.     

Insecurities of Women Regarding Breast Cancer Research: A Qualitative Study

Objectives

Only 1.2%–11% of all potential study participants participate in cancer studies. Low participation rates can result in bias or in a failure to obtain data saturation. Subject-scientific psychology assumes that reasons for acting are based on individual premises. The objective of this study was to render reproducible individual reasons of female breast cancer patients to participate or not participate in breast cancer studies using a qualitative approach.

Methods

Problem-based interviews were conducted with female breast cancer patients. The selection of interview partners continued until theoretical data saturation was achieved.

Results

As main arguments against participation emotional overload and too many medication side-effects were stated. Improvement of health-related values, long-term protection and comprehensive follow-up exams were stated as arguments for participation. Trust in the attending physician was mentioned as influencing both participation and non-participation.

Conclusions

A significant influential factor determining willingness to participate in studies was one''s contentment with patient-physician communication. In order to guarantee an adequate patient decision-making process, keeping existing standards for patient briefings is absolutely mandatory.     

The Effect of Breast‐Feeding with and without Formula Use on the Risk of Obesity at 4 Years of Age

Objective: To determine the minimal duration of breast‐feeding required to protect against later obesity, whether the concurrent use of formula lessened any protective effect of breast‐feeding, and what maternal or child characteristics might modify the association between breast‐feeding and child obesity. Research Methods and Procedures: This was a retrospective cohort study. Participants were 73, 458 white and black low‐income children followed from birth through 4 years of age. Obesity at age 4 years was defined as measured BMI ≥ 95th percentile. Feeding exposure was based on breast‐feeding duration and the age of formula initiation. Covariates were obtained from the children's birth certificates. Results: At age 4 years, the prevalence of obesity was 11.5%. Only 16% of children were breast‐fed 8 weeks or longer. Breast‐feeding was associated with a reduced risk of obesity only in white children whose mothers had not smoked in pregnancy. In this subgroup, the reduction in obesity risk (adjusted odds ratio, 95% confidence interval), compared with those never breast‐fed, occurred only for children who were breast‐fed at least 16 weeks without formula (0.71, 0.56 to 0.92) or at least 26 weeks with concurrent formula (0.70, 0.61 to 0.81). Among whites whose mothers smoked in pregnancy and among blacks, breast‐feeding was not associated with a reduced risk of obesity at age 4 years. Discussion: In a population of low‐income children, breast‐feeding was associated with a reduced risk of obesity at age 4 years only among whites whose mothers did not smoke in pregnancy and only when breast‐feeding continued for at least 16 weeks without formula or at least 26 weeks with formula.     

Targeting Hsp90 prevents escape of breast cancer cells from tyrosine kinase inhibition

Recent studies have identified development of resistance to tyrosine kinase inhibition (TKI) as a significant roadblock to effective treatment. One mechanism of resistance recently appreciated involves 'oncogene switching', or the re-activation of signaling pathways by one or more redundant upstream activators. In breast cancer models, ErbB TKIs such as gefitinib have been shown to lose the ability to modulate ErbB-driven signaling pathways over time, even though ErbB inhibition is maintained. Although incomplete ErB inhibition has been proposed to underlie this phenomenon, our findings suggest that oncogene switching can also re-activate downstream signaling pathways in breast cancer cells, even when ErbB inhibition is complete. We find that ErbB TKI-induced Src activation mediates downstream signaling rebound in SKBR3 cells, and we show that combination of Src and ErbB inhibitors is more effective and longlasting than is either TKI alone. Finally, the Hsp90 inhibitor 17-AAG, by simultaneously and durably inhibiting multiple signaling activators including ErbB and Src kinases, does not permit oncogene switching and results in a more prolonged and robust inhibition of downstream signaling pathways in breast cancer cells than do individual TKIs. These data support the continued clinical evaluation of Hsp90 inhibitors in breast cancer.     

Breast feeding and risk of breast cancer in young women. United Kingdom National Case-Control Study Group.

OBJECTIVE--To investigate whether breast feeding is related to subsequent risk of breast cancer. DESIGN--Population based case-control study designed primarily to investigate the relation between oral contraceptives and risk of breast cancer; data obtained from questionnaires administered by interviewers, general practitioner notes, and family planning clinic records. SETTING--11 health regions in Britain. SUBJECTS--Women diagnosed with breast cancer before age 36 living in the defined study areas. One control per case, matched for age, was selected from the list of the case''s general practitioner. 755 case-control pairs were interviewed. MAIN OUTCOME MEASURES--Duration of breast feeding each liveborn infant; timing of return of menses; hormone use; other risk factors for breast cancer. RESULTS--Risk of breast cancer fell with increasing duration of breast feeding (relative risk = 0.94 per three months'' breast feeding; test for trend p = 0.026) and with number of babies breast fed (relative risk = 0.86; test for trend, p = 0.017). Breast feeding each baby for longer than three months conferred no additional benefit. Breast feeding was more strongly negatively associated with risk of breast cancer than duration of postpartum amenorrhoea (chi 2 test for trend, p = 0.69). Hormonal suppression of lactation was unrelated to risk of breast cancer (relative risk = 0.96 per episode of suppressed lactation; test for trend, p = 0.72). CONCLUSIONS--These results suggest that breast feeding protects against the development of breast cancer in young women.     

Continuous Intragastric Milk Feeds in Infants of Low Birth Weight

In a feeding trial 66 infants of low birth weight received continuous intragastric milk feeds from the fourth hour of life, starting with 60 ml/kg/24 hr and reaching a maximum of 300 ml/kg/24 hr on the ninth day. Each infant received only full-strength milk, which was either expressed human breast milk or SMA-S26 (a proprietary low-protein adapted cows'' milk) or half-cream Regal milk (partly-skimmed evaporated cows'' milk). For various reasons 10 babies had to be withdrawn, and the final assessment was made on the 56 who completed the trial successfully.Persistent vomiting was a problem in only four infants. In two of them the trial was continued after gastric lavage and in the other two vomiting stopped when the volume was reduced. Despite a careful search no evidence was found of aspiration of feeds in any infant. Continuous intragastric milk infusion was shown to be a safe method of feeding infants of low birth weight and SMA-S26 was almost as well tolerated as human milk. Because of the high-protein content of half-cream cows'' milk preparations and the resultant high plasma amino-acid levels when they are given in these large volumes they should be avoided for this type of feeding although they produce better weight gains in the first week of life.     

Effects of haloperidol and clotrimazole on synaptic transmission in and contractile activity of smooth muscles of the guinea-pig intestine

In muscle strips of the guinea-pig large intestine, haloperidol and clotrimazole increased spontaneous electrical and contractile activities and decreased ATP-evoked hyperpolarization of smooth-muscle cells and the amplitude of inhibitory synaptic potentials. The pattern of effects of haloperidol on hyperpolarization induced by intramural stimulation of muscle strips was preserved under conditions of pre-incubation of the preparations in Krebs solutions containing pyridoxal-5′-phosphate, Nω-nitro-L-arginine, or apamin, as well as both apamin and tetraethylammonium. Neirofiziologiya/Neurophysiology, Vol. 39, Nos. 4/5, pp. 412–415, July–October, 2007.     

Current Concepts of Cancer Chemotherapy

The most impressive regressions obtained to date with the use of chemotherapy are in metastatic choriocarcinoma of females. In the management of Hodgkin''s disease, leukemia, and lymphoma, chemotherapy is a useful and accepted form of therapy. In many cases radiation and chemotherapy are interdependent. In disseminated carcinomas arising in the lung, ovary or breast, there is less likelihood of significant improvement with the use of chemotherapy, but if the disease is not amenable to radiotherapy, useful palliation can be obtained at times.While newer chemotherapeutic agents for cancer, notably the pyrimidine analogues, have a broader spectrum of antitumor effects than others investigated earlier, they should be regarded as providing a valuable research stimulus in the continued search for more effective and less toxic agents.     

High-performance liquid chromatographic method for the detection and quantitation of haloperidol and seven of its metabolites in microsomal preparations

An isocratic high-performance liquid chromatographic (HPLC) system was developed to analyze haloperidol and its potential metabolites. These compounds included 4-(4-chlorophenyl)-4-hydroxypiperidine (CPHP), haloperidol N-oxide (HNO), reduced haloperidol (RHAL), the 1,2,3,6-tetrahydropyridine analogue and its N-oxide, and the pyridinium ion from haloperidol (HP⁺). The HPLC system comprised a Hypersil CPS5 column with a mobile phase of acetonitrile (67%) and ammonium acetate (final concentration 10 mM) which was adjusted to pH 5.4 by acetic acid. The solvent was delivered at 1 ml/min. RHAL and CPHP were determined by an ultraviolet detector at 220 nm with a detection limit of 1 nmol/ml. All other compounds were determined at 245 nm and had a detection limit of 0.3 nmol/ml. This system was used to analyze a microsomal metabolic mixture of haloperidol. It was found that all above compounds except HNO were metabolites of haloperidol. In addition, two other metabolites were also well separated in this HPLC system which are proposed to be oxygenated haloperidol and the pyridone analogue of haloperidol. The HPLC system was used to carry out quantitative metabolic studies of haloperidol. It was found that the metabolism of haloperidol exhibits large inter-species differences. The apparent enzyme kinetic parameters were also determined using mice microsomes.     

The interplay between glucose and fatty acids on tube formation and fatty acid uptake in the first trimester trophoblast cells,HTR8/SVneo

Anillin (ANLN), an actin-binding protein, is required for cytokinesis. Recently, ANLN has been identified as a biomarker in diverse human cancers; however, the precise role of ANLN in breast cancer remains unclear. In this study, we firstly detected the expression of ANLN in 71 patients with breast cancer by immunohistochemistry, and found ANLN was highly expressed in breast cancer tissues. To evaluate the function of ANLN in breast cancer cells, we employed lentivirus-mediated RNA interference to knock down ANLN expression in two human breast cancer cell lines, MDA-MB-231, and ZR-75-30. Knockdown of ANLN remarkably inhibited the proliferation rate and colony formation ability of both breast cancer cell lines. Moreover, flow cytometry analysis showed that depletion of ANLN in MDA-MB-231 cells blocked the cell cycle progression, with more cells delayed at G₂/M phase, due to phosphorylation of Cdc2 and suppression of Cyclin D1. Furthermore, knockdown of ANLN strongly suppressed the migration of breast cancer cells, strengthening the evidence that ANLN could be involved in breast cancer progression. Our results may suggest ANLN as a potential target candidate in breast cancer.     

The social origins of infantile colic: questionnaire study covering 76,747 infants.

OBJECTIVE: To describe risk factors for infantile colic. DESIGN: Questionnaire administered by health visitors. SETTING: Sheffield. SUBJECTS: Mothers of 76,747 infants born between 1 August 1975 and 31 May 1988, interviewed when the infant was 1 month old. MAIN OUTCOME MEASURES: Reporting of infantile colic and its duration; weight of infant leeding, state of the home, socioeconomic characteristics of the parents, parents'' age, and mother''s parity. RESULTS: The odds of reporting infantile colic were increased with breast feeding (odds ratio of breast v bottle feeding 1.35 (95% confidence interval 1.28 to 1.43)), increasing parental age, lower parity, increasing parental age at leaving full time education, and more affluent homes and districts of residence. In a logistic regression analysis, mother''s age and parity and socioeconomic factors remained the most important risk factors for the reporting of infantile colic (each P < 0.005), and the effect of breast feeding was attenuated (odds ratio of breast v bottle feeding 1.09 (1.02 to 1.15)). CONCLUSION: At a population level, dietary factors contribute little to mothers'' reporting of infantile colic, and dietary change should not be the primary intervention.     

Epigenetic marks in estrogen receptor alpha CpG island correlate with some reproductive risk factors in breast cancer

Reproductive backgrounds, such as age at menarche and menopause, age of first full-term pregnancy (FFTP), number of full-term deliveries and oral contraceptive use are main hormone-related risk factors of breast cancer. It seems that the mentioned factors may affect the risk of breast cancer by enhancing the duration of exposure to estrogen as a potent carcinogen for breast tissue, but the molecular mechanism which links each risk factor to breast cancer is unclear. Estrogen mainly works via its nuclear receptor (ERα). As epigenetic alterations such as CpG methylation are potential links between endogenous or exogenous exposures and genome, we hypothesized that hormone-related risk factors may correlate with the epigenetic marks of the ERα promoter in breast tumors. In the present study, the CpG methylation status of the ERα gene in 99 samples of breast tumors belonged to women with different reproductive histories was evaluated. The reproductive history data were collected from patients. ERα CpG methylation was investigated by methylation specific PCR in DNA samples were obtained from the breast tumors. We could show that some of the hormone-related risk factors (early FFTP and increased number of pregnancies) were inversely correlated with epigenetic marks in ERα gene in breast tumors. Other hormone-related risk factors such as age of menarche and menopause and oral contraceptive use did not show any association with ERα methylation. It seems that pregnancy-related risk factors in comparison with other hormone-related factors work via different mechanism. As ERα methylation is a poor prognosis marker in breast tumors, its association with some modifiable reproductive risk factors (FFTP age and numbers of pregnancies) reiterates the importance of programming reproductive life style not only for prevention of breast cancer but also in favoring the prognosis of the affected women. The exact molecular mechanisms of the observed correlation need more investigation in the future.     

Haloperidol and Cerebral Metabolism in the Conscious Rat: Relation to Pharmacokinetics

The time course and distribution of alterations in cerebral metabolic activity after haloperidol administration were evaluated in relation to the pharmacokinetics of haloperidol and the topography of the dopaminergic system in the brain. Local cerebral glucose utilization was measured, using the 2-deoxyglucose technique, in awake rats after i.p. administration of the dopamine antagonist haloperidol (0.5 or 1 mg/kg). Haloperidol significantly reduced glucose utilization in 60% of 59 brain regions examined, but produced a large increase in the lateral habenula. The regional distribution of changes in glucose utilization was not closely related to the known anatomy of the brain dopaminergic system. The time course of the effect of haloperidol on cerebral metabolism was different for the two doses studied (0.5 and 1 mg/kg), and was not simply related to estimated brain concentrations of haloperidol. However, a linear relation between the metabolic effect and the time-integrated brain concentration was demonstrated. These results show that haloperidol has an effect on CNS metabolic activity that is more widespread than would be predicted from the topography of the dopaminergic system; this may be due to indirect propagation of the primary effects of haloperidol. The metabolic response to haloperidol depends on brain concentration and duration of exposure to the drug.     

Tourette's syndrome: a treatable tic.

Tourette''s syndrome, or Gilles de la Tourette''s disease, is a disorder characterized by involuntary tic-like muscular movements, compulsive behaviour and involuntary vocalization of sounds, words or profanities. It begins in childhood and may persist for life, with a varied pattern and course. Recent studies indicate an organic basis for the disorder, and an abnormality of dopamine or purine metabolism has been suggested. The treatment of choice is haloperidol administration; most patients do well with low or moderate doses for long periods. Because these patients are often mistakenly regarded as anxious, psychoneurotic or hysterical, correct diagnosis is important if they are to be treated appropriately and regarded properly in the home, school and society.     

女性乳房血供及神经支配的应用解剖进展

乳房作为女性身体的一个重要器官,不仅具有孕育生命的作用,更是女性形体美最显著的标志。然而由于各种原因导致女性乳房的形态千差万别,这不仅严重影响了女性的形体美,而且会对女性带来各种轻重程度不等的身体不适,更严重者甚至会对女性的心理健康造成极大的影响。这就需要整形外科医生通过乳房整形手术重新塑造女性乳房的形态美。由于术后易发生皮瓣及乳头乳晕血供障碍、乳头乳晕区域支配神经的损伤以及乳房形态不佳,使得巨乳缩小整形术具有手术难度大,手术风险高,术后患者满意度较差的特点,成为乳房整形手术中的一大难点。究其原因,我们对于女性乳房的血供和神经支配,尤其是乳房内血管神经的具体走行和分支仍然未能了解得十分清晰。因此,开展乳房血供及神经支配的应用解剖研究,对于改良目前的乳房整形手术特别是巨乳缩小整形术术式,以降低手术相关并发症的发生率,具有及其重要的临床指导意义。本文主要从乳房的血供和神经支配两个方面,对乳房的应用解剖进展进行综述。     

Plasma free MHPG and neuroendocrine responses to challenge doses of clonidine in Tourette's Syndrome: Preliminary report

The metabolic response to clonidine was examined in a pilot study of several patients with Tourette's Syndrome (TS). Preliminary results in a small number of patients suggested hypotheses for further investigation. Plasma free MHPG was reduced in three clinically responsive patients following a challenge dose of clonidine or haloperidol. During maintenance treatment with clonidine, a challenge dose of clonidine continued to elicit a decrease in plasma free MHPG, and CSF free MHPG was reduced. On the other hand, a clinically non-responsive TS patient on maintenance clonidine failed to show this decrease in plasma free MHPG. Prolactin, TSH, cortisol, T₃, and thyroxine indices were not changed by clonidine administration to TS patients. There was a variable GH response to clonidine. Neuroendocrine responses of patients to single oral doses of clonidine may provide a useful method of differentiating subgroups of childhood neuropsychiatric patients.     

What do women know about breast cancer prophylaxis and a healthy style of life?

AimThe aim of the study was to determine the factors influencing women's knowledge concerning breast cancer prophylaxis and find out the sources of the knowledge.BackgroundIn the Greater Poland region, breast cancer has been the most frequently detected tumour for years. The percentage of breast cancer cases has increased by 31% in the last decade.Materials and methodsThe study encompassed 337 women aged 40–59 who participated in the mammographic examinations. An original research tool was used which assessed the level of knowledge concerning breast cancer prophylaxis, the knowledge of health-oriented behaviour in this regard and the influence of the medical personnel on women's education.ResultsAge is a factor diversifying the knowledge of the breast self-examination method. Doctors and nurses were rarely indicated as a source of knowledge concerning breast cancer prophylaxis. The subjects presented a high level of knowledge of the factors increasing the risk of developing cancer.ConclusionsA correlation between the level of education and the knowledge of one's own breast to a degree which enables a woman to detect even a slight change was observed. Vital findings also concern the sources of knowledge concerning breast cancer prophylaxis. The results of the studies indicated little informative support on the part of the medical personnel; therefore, one should call for supplementing training courses for doctors and nurses focusing on the issues of prophylaxis, including the method of breast self-examination.     

Rare BRCA1 haplotypes including 3’UTR SNPs associated with breast cancer risk

Genetic markers identifying women at an increased risk of developing breast cancer exist, yet the majority of inherited risk remains elusive. While numerous BRCA1 coding sequence mutations are associated with breast cancer risk, BRCA1 mutations account for less then 5% of breast cancer risk. Since 3' untranslated region (3'UTR) polymorphisms disrupting microRNA (miRNA) binding can be functional and can act as genetic markers of cancer risk, we tested the hypothesis that such polymorphisms in the 3'UTR of BRCA1 and haplotypes containing these functional polymorphisms may be associated with breast cancer risk. We sequenced the BRCA1 3'UTR from breast cancer patients to identify miRNA disrupting polymorphisms. We further evaluated haplotypes of this region including the identified 3'UTR variants in a large population of controls and breast cancer patients (n=221) with known breast cancer subtypes and ethnicities. We identified three 3'UTR variants in BRCA1 that are polymorphic in breast cancer populations, and haplotype analysis including these variants revealed that breast cancer patients harbor five rare haplotypes not generally found among controls (9.50% for breast cancer chromosomes, 0.11% for control chromosomes, p=0.0001). Three of these rare haplotypes contain the rs8176318 BRCA1 3'UTR functional variant. These haplotypes are not biomarkers for BRCA1 coding mutations, as they are found rarely in BRCA1 mutant breast cancer patients (1/129 patients= 0.78%). These rare BRCA1 haplotypes and 3'UTR SNPs may represent new genetic markers of breast cancer risk.     

Mammographic Breast Density in Chinese Women: Spatial Distribution and Autocorrelation Patterns

Mammographic breast density (MBD) is a strong risk factor for breast cancer. The spatial distribution of MBD in the breast is variable and dependent on physiological, genetic, environmental and pathological factors. This pilot study aims to define the spatial distribution and autocorrelation patterns of MBD in Chinese women aged 40–60. By analyzing their digital mammographic images using a public domain Java image processing program for segmentation and quantification of MBD, we found their left and right breasts were symmetric to each other in regard to their breast size (Total Breast Area), the amount of BMD (overall PD) and Moran''s I values. Their MBD was also spatially autocorrelated together in the anterior part of the breast in those with a smaller breast size, while those with a larger breast size tend to have their MBD clustered near the posterior part of the breast. Finally, we observed that the autocorrelation pattern of MBD was dispersed after a 3-year observation period.