Occupational exposure to pesticides deregulates systemic cortisol levels in women with breast cancer and correlates with poor prognosis features.

Pesticides have been pointed out as hormone disruptors and may significantly affect the prognosis of hormone-dependent diseases such as breast cancer (BC). Here, we investigated the impact of occupational pesticide exposure on systemic cortisol levels in female rural workers diagnosed with BC. Occupational exposure was assessed by interviews with a standardized questionnaire. Plasma samples (112 from pesticide-exposed women and 77 from unexposed women) were collected in the afternoon, outside the physiological cortisol peak, and analyzed by a chemiluminescent paramagnetic immunoassay for the quantitative determination of cortisol levels in serum and plasma. The results from both groups were categorized according to patients' clinicopathological and exposure data. BC pesticide-exposed women presented higher levels of cortisol than the unexposed. Higher cortisol levels were also detected in the exposed group with more aggressive disease (triple-negative BC), with tumors over 2 cm, with lymph node metastases, and with high risk of disease recurrence and death. These findings demonstrated that there is an association between pesticide exposure and BC that affected cortisol levels and correlated to poor disease prognosis.

Occupational exposure to pesticides deregulates systemic cortisol levels in women with breast cancer and correlates with poor prognosis features

Pesticides have been pointed out as hormone disruptors and may significantly affect the prognosis of hormone-dependent diseases such as breast cancer (BC). Here, we investigated the impact of occupational pesticide exposure on systemic cortisol levels in female rural workers diagnosed with BC. Occupational exposure was assessed by interviews with a standardized questionnaire. Plasma samples (112 from pesticide-exposed women and 77 from unexposed women) were collected in the afternoon, outside the physiological cortisol peak, and analyzed by a chemiluminescent paramagnetic immunoassay for the quantitative determination of cortisol levels in serum and plasma. The results from both groups were categorized according to patients' clinicopathological and exposure data. BC pesticide-exposed women presented higher levels of cortisol than the unexposed. Higher cortisol levels were also detected in the exposed group with more aggressive disease (triple-negative BC), with tumors over 2 cm, with lymph node metastases, and with high risk of disease recurrence and death. These findings demonstrated that there is an association between pesticide exposure and BC that affected cortisol levels and correlated to poor disease prognosis.

Mini-Implants for Mandibular Overdentures: Cost-Effectiveness Analysis alongside a Randomized Trial.

Mandibular overdentures retained by 2 conventional implants have been considered the standard of care for complete edentulism, according to the McGill and York consensuses. However, many patients refuse this treatment modality due to the associated costs and postsurgical discomfort. Mini-implants have the chance to overcome these limitations due to their potentially lower costs and a relatively uncomplicated surgical technique. This study compared treatment costs and incremental cost-effectiveness following the insertion of mini-implants (2 or 4) or 2 standard-size implants for the retention of mandibular overdentures, by means of a randomized clinical trial. In total, 120 edentulous participants (mean age 59.5 ± 8.5 y) were randomly allocated into 3 groups according to treatment received: 4 mini-implants (group 1), 2 mini-implants (group 2), or 2 standard implants (group 3). Treatment costs and outcomes (Oral Health Impact Profile for Edentulous [OHIP-EDENT] and satisfaction with the dentures) were evaluated after 6 mo. Incremental cost-effectiveness ratios (ICERs) were calculated for each intervention in terms of cost per 1-point change in patient outcomes. A 1-way sensitivity analysis was performed considering a 95% confidence interval variation in cost and outcome parameters, represented in tornado diagrams. Overall treatment cost was the lowest for group 2 (average cost: US$318.08), followed by group 1 (US$510.75) and group 3 (US$566.13). Groups did not differ in terms of the length of unscheduled appointments and time spent by participants. In summary, our findings indicate that mandibular overdentures retained by 2 or 4 mini-implants are less costly compared to 2-implant overdentures. Despite the lower costs of overdentures retained by 2 mini-implants, those retained by 4 mini-implants showed further improvement in patient-reported outcomes and reduced costs compared to standard implants ( ClinicalTrials.gov NCT01411683). Knowledge Transfer Statement: This report shows that mini-implant retained overdentures are less costly than overdenture treatment on 2 standard-sized implants. Treatment with 2 mini-implants is an effective procedure to substantially save resources, whereas treatment with 4 mini-implants provides better results from a patient perspective combined with slightly reduced costs compared to the treatment with 2 standard implants. Therefore, mini-implant overdentures may be effective and more accessible than overdentures on 2 standard-size implants for those with limited incomes.

Mini vs. Standard Implants for Mandibular Overdentures: A Randomized Trial.

A mandibular implant-retained overdenture is considered a first-choice treatment for edentulism. However, some aspects limit the use of standard implants-for example, the width of edentulous ridges, chronic diseases, fear, or costs. This randomized trial compared mandibular overdentures retained by 2 or 4 mini-implants with standard implants, considering oral health-related quality of life (OHRQoL), patient satisfaction, and complications such as lost implant. In sum, 120 edentulous men and women (mean age, 59.5 ± 8.5 y) randomly received 4 mini-implants, 2 mini-implants, or 2 standard implants. Participants provided data regarding OHRQoL and satisfaction until 12 mo. Clinical parameters, including implant survival rate, were also recorded. Both 2 and 4 mini-implants led to better OHRQoL, compared with 2 standard implants. Treatment with 4 mini-implants was more satisfying than 2 standard implants, with 2 mini-implants presenting intermediate results. Implant survival rate was 89%, 82%, and 99% for 4 mini-implants, 2 mini-implants, or 2 standard implants, respectively. Overdentures retained by 4 or 2 mini-implants can achieve OHRQoL and satisfaction at least comparable with that of 2 standard implants. However, the survival rate of mini implants is not as high as that of standard implants (ClinicalTrials.gov NCT01411683).

Short-term post-operative pain and discomfort following insertion of mini-implants for retaining mandibular overdentures: a randomized controlled trial.

The retention of removable dentures by mini-implants is a relatively recent treatment modality and may lead to minimal post-operative trauma. This study compared post-operative pain and discomfort following the insertion of mini-implants (two or four) or two standard-size implants for the retention of mandibular overdentures. One hundred and twenty edentulous participants (mean age 59.5 ± 8.5 years) were randomly allocated into three groups according to received treatment: (GI) four mini-implants, (GII) two mini-implants or (GIII) two standard implants. Seven days after implant insertion, patients answered questions (100-mm VAS) relating to pain, swelling, and discomfort with chewing, speech and hygiene, considering their experiences during the 1st and 6th day. Groups were compared by two-way anova (α = 0.05). All participants (GI: 38; GII: 42; GIII: 40) were analysed after 7 days. At the 6th day, GI felt significantly higher pain than GII and GIII. GI also reported more difficulty in performing oral hygiene practices than GIII during the 1st day. There was no significant difference between groups for the other questions and periods. No participant suffered unexpected side effects. The use of four mini-implants induces more intense post-operative pain at the 6th day than the insertion of two mini- or conventional fixtures, as well as more difficult oral hygiene on the 1st day. Clinicaltrials.gov, NCT01411683; FAPESP, 2011/00688-7 and 2011/23347-0.

A randomised trial of a simplified method for complete denture fabrication: patient perception and quality.

Complete denture fabrication involves a series of complex technical procedures. Nevertheless, simplified methods may be as effective as conventional ones albeit the lesser use of time and resources, without disadvantage for the patient. This study compared a simplified method for complete denture fabrication to a conventional protocol in terms of oral health-related quality of life (OHRQoL), patient satisfaction and denture quality. Forty-two edentulous patients requesting treatment with complete dentures were randomly allocated into two study groups. Group S received dentures fabricated by a simplified method and Group C received conventionally fabricated dentures. Before interventions and after three and 6 months following insertion, OHRQoL and patient satisfaction were analysed by specific instruments. A prosthodontist assessed denture quality 3 months after delivery. Groups presented no difference for OHRQoL, denture quality and general satisfaction. Differences regarding patient satisfaction with some aspects of the dentures were found after 3 months (S > C), but were insignificant at 6 months. It was concluded that the simplified method is able to produce dentures of a quality comparable to those produced by the conventional method, influencing OHRQoL and patient satisfaction similarly.

A randomised trial on simplified and conventional methods for complete denture fabrication: masticatory performance and ability.

OBJECTIVES: To compare a simplified method to a conventional protocol for complete denture fabrication regarding masticatory performance and ability. METHODS: A sample was formed by edentulous patients requesting treatment with maxillary and mandibular complete dentures. Participants were randomly divided into two groups: Group S, which received dentures fabricated by a simplified method, and Group C (n=21 each), which received conventionally fabricated dentures. After three months following insertion, masticatory performance was evaluated by a colorimetric assay based on chewing two capsules as test food during twenty and forty cycles. Masticatory ability was assessed by a questionnaire with binary answers and a single question answered by means of a 0-10 scale. A third group (DN) formed by seventeen dentate volunteers served as an external comparator. Groups were compared by statistical tests suitable for data distribution (α=0.05). RESULTS: Thirty-nine participants were assessed for three months (twenty from Group C and nineteen from Group S). Groups C and S presented similar masticatory performance which corresponded to approximately 30% of Group DN. Results for masticatory ability showed similarity between S and C, regardless of the assessment method, although an isolate questionnaire item showed more favourable results for the first group. CONCLUSIONS: The simplified method for complete denture fabrication is able to restore masticatory function to a level comparable to a conventional protocol, both physiologically and according to patient's perceptions. CLINICAL SIGNIFICANCE: Although masticatory function is impaired by the loss of natural teeth and dentures can restore only a fraction of such function, patients can benefit from a simplified protocol for complete denture fabrication to the same extent they would by conventional techniques.

Microencapsulated citronella oil for mosquito repellent finishing of cotton textiles.

Microcapsules containing citronella essential oil were prepared by complex coacervation and applied to cotton textiles in order to study the repellent efficacy of the obtained fabrics. Citronella released from treated textiles was indirectly monitored by the extractable content of its main components. Repellent activity was assessed by exposure of a human hand and arm covered with the treated textiles to Aedes aegypti mosquitoes. Fabrics treated with microencapsulated citronella presented a higher and longer lasting protection from insects compared to fabrics sprayed with an ethanol solution of the essential oil, assuring a repellent effect higher than 90% for three weeks. Complex coacervation is a simple, low cost, scalable and reproducible method of obtaining encapsulated essential oils for textile application. Repellent textiles were achieved by padding cotton fabrics with microcapsules slurries using a conventional pad-dry method. This methodology requires no additional investment for textile finishing industries, which is a desirable factor in developing countries.

Exploratory factor analysis of the Brazilian OHIP for edentulous subjects.

The use of seven domains for the Oral Health Impact Profile (OHIP)-EDENT was not supported for its Brazilian version, making data interpretation in clinical settings difficult. Thus, the aim of this study was to assess patients' responses for the translated OHIP-EDENT in a group of edentulous subjects and to develop factor scales for application in future studies. Data from 103 conventional and implant-retained complete denture wearers (36 men, mean age of 69.1 +/- 10.3 years) were assessed using the Brazilian version of the OHIP-EDENT. Oral health-related quality of life domains were identified by factor analysis using principal component analysis as the extraction method, followed by varimax rotation. Factor analysis identified four factors that accounted for 63% of the 19 items total variance, named masticatory discomfort and disability (four items), psychological discomfort and disability (five items), social disability (five items) and oral pain and discomfort (five items). Four factors/domains of the Brazilian OHIP-EDENT version represent patient-important aspects of oral health-related quality of life.

Cholesterol induces fetal rat enterocyte death in culture.

The effect of cholesterol on fetal rat enterocytes and IEC-6 cells (line originated from normal rat small intestine) was examined. Both cells were cultured in the presence of 20 to 80 microM cholesterol for up to 72 h. Apoptosis was determined by flow cytometric analysis and fluorescence microscopy. The expression of HMG-CoA reductase and peroxisome proliferator-activated receptor gamma (PPARgamma) was measured by RT-PCR. The addition of 20 microM cholesterol reduced enterocyte proliferation as early as 6 h of culture. Reduction of enterocyte proliferation by 28 and 41% was observed after 24 h of culture in the presence and absence of 10% fetal calf serum, respectively, with the effect lasting up to 72 h. Treatment of IEC-6 cells with cholesterol for 24 h raised the proportion of cells with fragmented DNA by 9.7% at 40 microM and by 20.8% at 80 microM. When the culture period was extended to 48 h, the effect of cholesterol was still more pronounced, with the percent of cells with fragmented DNA reaching 53.5% for 40 microM and 84.3% for 80 microM. Chromatin condensation of IEC-6 cells was observed after treatment with cholesterol even at 20 microM. Cholesterol did not affect HMG-CoA reductase expression. A dose-dependent increase in PPARgamma expression in fetal rat enterocytes was observed. The expression of PPAR-gamma was raised by 7- and 40-fold, in the presence and absence of fetal calf serum, respectively, with cholesterol at 80 mM. The apoptotic effect of cholesterol on enterocytes was possibly due to an increase in PPARgamma expression.

Cholesterol induces fetal rat enterocyte death in culture

The effect of cholesterol on fetal rat enterocytes and IEC-6 cells (line originated from normal rat small intestine) was examined. Both cells were cultured in the presence of 20 to 80 æM cholesterol for up to 72 h. Apoptosis was determined by flow cytometric analysis and fluorescence microscopy. The expression of HMG-CoA reductase and peroxisome proliferator-activated receptor gamma (PPARgamma) was measured by RT-PCR. The addition of 20 æM cholesterol reduced enterocyte proliferation as early as 6 h of culture. Reduction of enterocyte proliferation by 28 and 41 percent was observed after 24 h of culture in the presence and absence of 10 percent fetal calf serum, respectively, with the effect lasting up to 72 h. Treatment of IEC-6 cells with cholesterol for 24 h raised the proportion of cells with fragmented DNA by 9.7 percent at 40 æM and by 20.8 percent at 80 æM. When the culture period was extended to 48 h, the effect of cholesterol was still more pronounced, with the percent of cells with fragmented DNA reaching 53.5 percent for 40 æM and 84.3 percent for 80 æM. Chromatin condensation of IEC-6 cells was observed after treatment with cholesterol even at 20 æM. Cholesterol did not affect HMG-CoA reductase expression. A dose-dependent increase in PPARgamma expression in fetal rat enterocytes was observed. The expression of PPAR-gamma was raised by 7- and 40-fold, in the presence and absence of fetal calf serum, respectively, with cholesterol at 80 mM. The apoptotic effect of cholesterol on enterocytes was possibly due to an increase in PPARgamma expression.

Cholesterol changes the fatty acid composition of rat enterocytes.

The effect of free cholesterol on the fatty acid composition and growth of rat fetal enterocytes was investigated in the absence and presence of 10% (v/v) fetal calf serum. Cholesterol caused a significant reduction of cell number after 6 and 12 h in culture. The fatty acid composition of enterocytes cultured in the presence of serum was also changed by the presence of 20 microM cholesterol. The fatty acid profile was determined by HPLC using fluorescence detection (325 nm excitation and 395 nm emission). Cholesterol (20 microM) increased the proportion (given in percentage of the total fatty acids) of the following fatty acids in cultured cells: lauric (by 42%), oleic (by 34%), linoleic (by 44%) and gamma-linolenic (by 20%) acids and reduced the proportion of palmitic (by 12%), stearic (by 20%), arachidonic (by 21%) and docosahexaenoic (by 44%) acids. In addition to modifying the content of individual fatty acids, cholesterol increased the polyunsaturated/saturated fatty acid ratio from 0.48 to 0.67 and the unsaturation index from 67.12 to 75.30. This is the first evidence that cholesterol modifies fatty acid composition possibly via de novo fatty acid synthesis and desaturation.

Cholesterol changes the fatty acid composition of rat enterocytes

The effect of free cholesterol on the fatty acid composition and growth of rat fetal enterocytes was investigated in the absence and presence of 10 percent (v/v) fetal calf serum. Cholesterol caused a significant reduction of cell number after 6 and 12 h in culture. The fatty acid composition of enterocytes cultured in the presence of serum was also changed by the presence of 20 æM cholesterol. The fatty acid profile was determined by HPLC using fluorescence detection (325 nm excitation and 395 nm emission). Cholesterol (20 æM) increased the proportion (given in percentage of the total fatty acids) of the following fatty acids in cultured cells: lauric (by 42 percent), oleic (by 34 percent), linoleic (by 44 percent) and gamma-linolenic (by 20 percent) acids and reduced the proportion of palmitic (by 12 percent), stearic (by 20 percent), arachidonic (by 21 percent) and docosahexaenoic (by 44 percent) acids. In addition to modifying the content of individual fatty acids, cholesterol increased the polyunsaturated/saturated fatty acid ratio from 0.48 to 0.67 and the unsaturation index from 67.12 to 75.30. This is the first evidence that cholesterol modifies fatty acid composition possibly via de novo fatty acid synthesis and desaturation

Acetate and propionate potentiate the antiproliferative effect of butyrate on RBL-2H3 growth.

1. The effect of acetate, propionate, and butyrate separately and combined on RBL-2H3 (a rat basophilic leukemic cell type) proliferation during 24, 48, and 72 hr was examined. Also, the effect of a mixture of the three volatile fatty acids on proliferation of HeLa-155 (a human adenocarcinoma), C57 B1/6J (a mouse melanoma), and MCF-7 (human breast tumor) during 8 days was investigated. 2. Acetate and propionate per se did not present any effect on RBL-2H3 growth during 72 hr, however, when acetate and propionate were added together a significant inhibition of this cell growth was found; 18% for 48 and 37% for 72 hr. The addition of butyrate to the culture medium caused a 75% decrease in the rate of this cell growth either after 48 and 72 hr. This effect of butyrate was pronounced by acetate (86% and 90% for 48 and 72 hr, respectively), propionate (87% for 48 and 93% for 72 hr), and acetate and propionate together (76% for 48 and 92% for 72 hr). 3. Daily addition of a mixture of the short-chain fatty acids (10 mM acetate, 2 mM propionate and 1.5 mM butyrate) markedly decreased the number of cells after 8 days: 58% for RBL-2H3, 42% for HeLa-155, 91% for C57 B1/6J and 55% for MCF-7. 4. These results support the proposition that a fiber-rich diet that leads to great production of butyrate but also of propionate and acetate would be more effective to prevent the occurrence of colorectal cancer than the administration of this short-chain fatty acid given alone.

Wheat bran- but not oat bran-enriched diets increase the mucosal height of the cecum and colon of newly weaned and aged rats.

The effect of diets enriched with oat or wheat bran (prepared by the addition of 300 g of each fiber to 1000 g of the regular diet), given for 8 weeks, on the mucosal height of the colon and cecum was investigated. Newly weaned (21 days old) and aged (12 months old) male Wistar rats were used in this study. As compared to controls, diets enriched with wheat bran provoked a significant increase in the mucosal height, whereas oat bran did not cause any effect. In newly weaned rats (21 days old), wheat bran increased the mucosal height (microns) in the cecum by 20% (mean +/- SEM for 8 rats; 169.1 +/- 5.2 and 202.9 +/- 8.0 for control and wheat bran, respectively) and in the colon (218.8 +/- 7.2 and 264.5 +/- 18.8 for control and wheat bran, respectively). A similar effect was observed in aged rats (12 months old), with an increase of 15% in the mucosal height (microns) of the cecum (mean +/- SEM of 8 rats; 193.2 +/- 8.6 and 223.7 +/- 8.3 for control and wheat bran, respectively) and of 17% in the colon (300.4 +/- 9.2 and 352.2 +/- 15.9 for control and wheat bran, respectively).

Wheat bran -but not oat bran- enriched diets increase the mucosal height of the cecum and colon of newly weaned and aged rats

The effect of diets enriched with oat or wheat bran (prepared by the addition of 300 g of each fiber to 1000 g of the regular diet), given for 8 weeks, on the mucosal height of the colon and cecum was investigated. Newly weaned (21 days old) and aged (12 months old) male Wistar rats were used in this study. As compared to controls, diets enriched with wheat bran provoked a significant increase in the mucosal height, whereas oat bran did not cause any effect. In newly weaned rats (21 days old), wheat bran increased the mucosal height (mum) in the cecum by 20 per cent (mean + SEM for 8 rats; 169.1 + 5.2 and 202.9 + 8.0 for control and wheat bran, respectively) and in the colon (218.8 + 7.2 and 264.5 + 18.8 for control and wheat bran, respectively). A similar effect was observed in aged rats (12 months old), with an increase of 15 per cent in the mucosal height (mum) of the cecum (mean + SEM of 8 rats; 193.2 + 8.6 and 223.7 + 8.3 for control and wheat bran, respectively) and of 17 per cent in the colon (300.4 + 9.2 and 352.2 + 15.9 for control and wheat bran, respectively).

Fatty acid composition of lymphocytes and macrophages from rats fed fiber-rich diets: a comparison between oat bran- and wheat bran-enriched diets.

The effect of oat bran- (OBD) and wheat bran-enriched diets (WBD) on fatty acid composition of neutral lipids and phospholipids of rat lymphocytes and macrophages was investigated. In neutral lipids of lymphocytes, OBD reduced the proportion of palmitoleic acid (48%), whereas WBD reduced by 43% palmitoleic acid and raised oleic (18%), linoleic (52%), and arachidonic (2.5-fold) acids. In neutral lipids of macrophages, OBD increased palmitic (16%) and linoleic (29%) acids and slightly decreased oleic acid (15%). The effect of WBD, however, was more pronounced: It reduced myristic (60%), stearic (24%) and arachidonic (63%) acids, and it raised palmitic (30%) and linoleic (2.3-fold) acids. Neither OBD nor WBD modified the composition of fatty acids in phospholipids of lymphocytes. In contrast, both diets had a marked effect on composition of fatty acids in macrophage phospholipids. OBD raised the proportion of myristic (42%) and linoleic (2.4-fold) acids and decreased that of lauric (31%), palmitoleic (43%), and arachidonic (29%) acids. WBD increased palmitic (18%) and stearic (23%) acids and lowered palmitoleic (35%) and arachidonic (78%) acids. Of both cells, macrophages were more responsive to the effect of the fiber-rich diets on fatty acid composition of phospholipids. The high turnover of fatty acids in macrophage membranes may explain the differences between both cells. The modifications observed due to the effects of both diets were similar in few cases: an increase in palmitic and linoleic acids of total neutral lipids occurred and a decrease in palmitoleic and arachidonic acids of phospholipid. Therefore, the mechanism involved in the effect of both diets might be different.

Atrial natriuretic peptide modulates the effect of angiotensin II on the concentration of free calcium in the cytosol of Mandin-Darby canine kidney cells.

The effect of angiotensin II (ANG II) and atrial natriuretic peptide (ANP) on intracellular free calcium concentration [Ca2+]i was investigated in Mandin-Darby canine kidney (MDCK) cells in culture. Changes in [Ca2+]i were monitored fluorometrically with the Ca(2+)-sensitive probe fura-2/AM at 37 degrees C using a Perkin-Elmer LS-5 spectrofluorimeter (excitation 340/380 nm, slit 3 nm; emission 520 nm, slit 10 nm). MDCK cells exhibited a mean baseline [Ca2+]i of 98 +/- 10 nM. The addition of increasing concentrations of ANG II (1 pM to 1 microM) to the cell suspension led to a progressive increase in [Ca2+]i to 2-3 times basal levels. In contrast, addition of 1 microM ANP to the cell suspension led to a very rapid 60% decrease in [Ca2+]i. The addition of 1 pM to 1 microM ANG II immediately after 1 microM ANP caused an increase in [Ca2+]i which never exceeded the basal level in the absence of ANP. The data indicate that ANG II increases cell [Ca2+]i, as expected, and provide the new observation that ANP reduces [Ca2+]i in these cells. Furthermore, ANP reduces the increase in [Ca2+]i elicited by ANG II, thus modulating the effect of ANG II on [Ca2+]i.

Atrial natriuretic peptide modulates the effect of angiotensin II on the concentration of free calcium in the cytosol of Mandin-Darby canine kidney cells

The effect of angiotensin II (ANG II) and atrial natriuretic peptide (ANP) on intracellular free calcium concentration [ Ca²+]i was investigated in Mandin-Darby canine kidney (MDCK) cells in culture. Changes in [Ca²+]i were monitored fluorometrically with the Ca²+ -sensitive probel fura -2/AM at 37ºC using Perkin-Elmer LS-5 spectrofluorimeter (excitation 340/380 nm,slite 3 nm; emission 520 nm, slit 10 nm). MDCK cells exhibited a mean baseline [Ca²+]i of 98 ñ 10 nM. the addition of increasing concentrations of SNG II (1 pM to 1 µM) to the cell suspension led to a progressive increase in [Ca²+]i to 2-3 times basal levels. In contrast, addition of 1 µM ANP to the cell suspension led to a very rapid 60 percent decrease in [Ca²+]i. The addition of 1 pM to 1 µM ANG II immediately after 1 µM ANP caused an increase in [Ca²+]i which never exceded the basal level in the absence of ANP. The data indicate that ANG II increases cell [Ca²+]i as expected, and provide the new observation that ANP reduces [Ca²+]i in these cells. Further more, ANP reduces the increase in [Ca²+]i elicited by ANG II, thus modulating the effect of ANG II on [Ca²+]i