Variants in Pharmacokinetic Transporters and Glycemic Response to Metformin: A Metgen Meta-Analysis.

Therapeutic response to metformin, a first-line drug for type 2 diabetes (T2D), is highly variable, in part likely due to genetic factors. To date, metformin pharmacogenetic studies have mainly focused on the impact of variants in metformin transporter genes, with inconsistent results. To clarify the significance of these variants in glycemic response to metformin in T2D, we performed a large-scale meta-analysis across the cohorts of the Metformin Genetics Consortium (MetGen). Nine candidate polymorphisms in five transporter genes (organic cation transporter [OCT]1, OCT2, multidrug and toxin extrusion transporter [MATE]1, MATE2-K, and OCTN1) were analyzed in up to 7,968 individuals. None of the variants showed a significant effect on metformin response in the primary analysis, or in the exploratory secondary analyses, when patients were stratified according to possible confounding genotypes or prescribed a daily dose of metformin. Our results suggest that candidate transporter gene variants have little contribution to variability in glycemic response to metformin in T2D.

Initiation of glucose-lowering treatment decreases international normalized ratio levels among users of vitamin K antagonists: a self-controlled register study.

UNLABELLED: ESSENTIALS: It is not known if initiation of glucose-lowering drugs alters the efficacy of vitamin K antagonists (VKA). We examined if glucose-lowering drugs affected international normalized ratio (INR) in VKA-treated patients. Upon initiating glucose-lowering drugs, 51% of patients had INR values below the therapeutic window. Monitoring of INR levels should be intensified upon initiation of glucose-lowering drugs. BACKGROUND: It is not known whether initiation of antidiabetic treatment affects the effect of vitamin K antagonists (VKAs). It was previously shown that metformin affects the effect of one VKA, phenprocoumon. OBJECTIVES: The aim of this study was to determine if initiation of glucose-lowering treatment affects the international normalized ratio (INR) and dose requirements of the anticoagulant VKAs warfarin and phenprocoumon. PATIENTS/METHODS: We performed a self-controlled retrospective register-based study. A total of 118 patients commencing glucose-lowering treatment while being treated with warfarin or phenprocoumon were included in the study. We compared INR, dose/INR and proportion of patients with at least one sub-therapeutic INR measurement before and after initiation of glucose-lowering treatment. RESULTS: Initiation of glucose-lowering treatment caused mean INR to decrease from 2.5 to 2.2 (decrease of -0.3 [95% CI: -0.1; -0.5]) and led to more than half of the patients having at least one sub-therapeutic INR measurement. Six to 12 weeks later, the VKA dose/INR was increased by 11%, indicating a weakened effect of the VKA. CONCLUSION: Initiation of glucose-lowering treatment reduces the anticoagulant effect of VKAs to an extent that is likely to be clinically relevant. This finding needs confirmation and mechanistic explanation.

Trends in colorectal cancer survival in northern Denmark: 1985-2004.

OBJECTIVE: The prognosis for colorectal cancer (CRC) is less favourable in Denmark than in neighbouring countries. To improve cancer treatment in Denmark, a National Cancer Plan was proposed in 2000. We conducted this population-based study to monitor recent trends in CRC survival and mortality in four Danish counties. METHOD: We used hospital discharge registry data for the period January 1985-March 2004 in the counties of north Jutland, Ringkjøbing, Viborg and Aarhus. We computed crude survival and used Cox proportional hazards regression analysis to compare mortality over time, adjusted for age and gender. A total of 19,515 CRC patients were identified and linked with the Central Office of Civil Registration to ascertain survival through January 2005. RESULTS: From 1985 to 2004, 1-year and 5-year survival improved both for patients with colon and rectal cancer. From 1995-1999 to 2000-2004, overall 1-year survival of 65% for colon cancer did not improve, and some age groups experienced a decreasing 1-year survival probability. For rectal cancer, overall 1-year survival increased from 71% in 1995-1999 to 74% in 2000-2004. Using 1985-1989 as reference period, 30-day mortality did not decrease after implementation of the National Cancer Plan in 2000, neither for patients with colon nor rectal cancer. However, 1-year mortality for patients with rectal cancer did decline after its implementation. CONCLUSION: Survival and mortality from colon and rectal cancer improved before the National Cancer Plan was proposed; after its implementation, however, improvement has been observed for rectal cancer only.

[Marginal fit of dental restorations and the periodontium in Swiss army recruits]. / Restaurationsrandschlusse und Parodont bei Schweizer Rekruten.

The present study analyzed the relationship between periodontal health adjacent to filled and unfilled tooth sites in young men (recruits). The status of oral health of 419 Swiss army recruits, aged 19 to 20 years was assessed by determining Plaque Index (PI), Retention Index (RI) and Gingival Index (GI) as well as Pocket Probing Depth (PPD) and Probing Attachment Loss (PAL). In addition, the level of alveolar bone was measured using digitized bite-wing radiographs with an enlargement of 4.5x. Filling margins were assessed and the distance between the alveolar bone crest and the cemento-enamel junction (CEJ) measured to the nearest one tenth of a millimeter. These data were compared with the clinical parameters. A total of 8'050 sites were examined. 765 or 9.5 of the sites in the posterior area were filled. 119 of them showed filling overhangs larger than 0.2 mm. Thus, 1.5 % of the examined sites had a significant overhanging margin. All clinical parameters had greater values at filled than at unfilled sites. The differences were statistically not significant. Even the sites with margins overhanging more than 0.8 mm (n=14) did not show significantly different parameters compared to unfilled sites. The comparison with a similar study involving recruits 11 years earlier assessed that the recruits of 1996 had less and smaller filling overhangs. This, in turn, means that, in Switzerland restorative dentistry in young males has been markedly improved during the 1980's and 1990's.

Comparison of the clinical effects and gingival abrasion aspects of manual and electric toothbrushes.

AIMS: The clinical effects and gingival abrasion aspects of 2 electrical tooth-brushes (Braun Oral-B Plak Control Ultra and the novel development Braun Oral-B Plak Control 3D) were to be compared with conventional manual tooth-brushing. MATERIAL AND METHODS: In a cross-over study, 26 dental student volunteers participated and were assigned to 1 of 3 groups. Following instruction in the use of the electric as well as manual toothbrushes, the volunteers were timed for 2 min each day to apply one electric or the manual toothbrush, respectively, during 3 experimental phases of 2 weeks. No other methods of tooth cleaning were to be performed except the one specified for the respective test period. When brushing manually, the Bass toothbrushing technique was applied. Between each test period, a recovery period of 1 week was allowed during which no oral hygiene was performed at all. At the start and the end of each of the experimental periods, the extension of plaque deposits from the gingival margin in coronal direction was assessed using the Turesky et al. modification of the Quigley and Hein plaque index. Presence or absence of gingival inflammation was evaluated by bleeding and probing (BOP). The extent and severity of gingival abrasions were assessed by use of a modified method of Breitenmoser et al. and adapted by Danser et al. RESULTS: The plaque-reducing effect was similar in all groups with the same cleaning regime. For that reason, the result of the different experimental phases with the respective cleaning modalities were collapsed. Cleaning with the Braun Oral-B Plak Control Ultra electric toothbrush resulted consistently in the lowest plaque scores when compared to both the Braun Oral-B Plak Control 3D and the manual toothbrush. Although the differences in plaque reduction were statistically significant between cleaning with Braun Oral-B Plak Control Ultra and 3D, they were small and of questionable clinical relevance. No significant differences in plaque reductions were found between manual brushing and any of the 2 electric brushes. Gingival abrasions were least pronounced following brushing with the Braun Oral-B Plak Control 3D electric toothbrush. However, no significant differences in gingival abrasion were encountered following brushing with the Braun Oral-B Plak Control Ultra electric in comparison with the manual toothbrush. CONCLUSIONS: The results of the present study have shown that in a group of dental students trained in manual brushing technique, where efficacy was similar with the 3 toothbrushes tested, there is no evidence of greater gingival abrasion with either Braun Oral-B Plak Control Ultra or 3D when compared with a manual brush.

Histochemical tracing of bismuth in testis from rats exposed intraperitoneally to bismuth subnitrate.

The histochemical silver amplification technique autometallography (AMG), was used to trace bismuth in the testis of Wistar rats injected intraperitoneally with bismuth subnitrate. In the seminiferous tubules, bismuth was located in lysosomes of Sertoli cells closely associated with heads of spermatids in the late stages of the spermatogenesis, i.e. shortly before the release of Step 19 spermatids in Stage XIII. No bismuth-specific AMG silver grains were detected in the spermatogenic cell line. However, tails of free sperm cells located in the tubular lumen showed autometallographic grains in close contact to the nine outer microtubule doublets in the axonema. Leydig cells concentrated huge amounts of AMG-bismuth in their lysosomes. Furthermore, parallel exposure to selenium significantly increased the amount of histochemically traceable bismuth in the rat testis.

Dietary structured triacylglycerols containing docosahexaenoic acid given from birth affect visual and auditory performance and tissue fatty acid profiles of rats.

To examine whether it is possible to enhance the level of 22:6(n-3) in the central nervous system, newborn rats were fed dietary supplements containing oils with either specific or random triacylglycerol structure, but similar concentrations of polyunsaturated fatty acids. In the specific structured oil, 22:6(n-3) was located in the sn-2 position, whereas it was equally distributed among the three positions in the triacylglycerol molecule in the randomized oil. A reference group was fed rat milk before weaning and nonpurified diet after weaning. After 12 wk, the levels of 22:6(n-3) in brain and liver phospholipids were higher in the groups fed the experimental diets than in the reference group. The specific structured oil resulted in the highest level of 22:6(n-3) in the brain, whereas the level of 22:6(n-3) was highest in the liver of the group fed randomized oil, indicating differences in metabolism of fatty acids resulting from their position in the dietary triacylglycerol molecule. The higher levels of 22:6(n-3) were accompanied by significantly lower levels of the long-chain (n-6) polyunsaturated fatty acids compared with the reference group. The fatty acid profiles, including the level of 22:6(n-3), in the retina phospholipids were not affected by the three different diets apart from a lower level of 20:4(n-6) in rats fed the experimental diets, indicating a strong tendency to maintain a high level of 22:6(n-3) in the retina. The changes in the fatty acid profiles did not result in differences in learning ability, but caused changes in visual function, evidenced by higher latency of the b-wave and lower oscillatory potential, and in auditory brainstem response, evidenced by generally greater amplitude of wave Ia in the group fed specific structured oil.

Early dietary intervention with structured triacylglycerols containing docosahexaenoic acid. Effect on brain, liver, and adipose tissue lipids.

Newborn rats were fed liquid diets containing 7 wt% fat in which 3.8% of the total fatty acids were 22:6n-3. The fats were either a specific structured oil with 22:6n-3 mostly located in the sn-2 position or a randomized oil with 22:6n-3 equally distributed in the triacylglycerol (TAG) molecules. The oils were manufactured by interesterification of fish oil TAG with free fatty acids from butterfat. The pups were tube-fed three times a day and stayed with their dams during the night. After 14 d they were fed solid diets containing the same oils for the next 7 d. A reference group stayed with the dams and received ordinary rat chow at weaning. In general no significant differences between the two dietary treatments were observed in the tissues examined except for adipose tissue. The levels of 22:6n-3 were significantly increased in brain phosphatidylcholines (PC) and phosphatidylserines (PS) of both experimental groups compared with the reference group after three weeks, whereas no differences were found in brain phosphatidylethanolamines (PE) and phosphatidylinositols (PI). In all groups and all phospholipids examined, the levels of 20:4n-6 generally decreased from 1 to 3 wk and were significantly lower in the experimental groups compared with the reference group at 3 wk except for PI. In liver, PC and PE 22:6n-3 remained constant in the experimental groups but decreased significantly in the reference group, whereas in liver PS 22:6n-3 increased in all groups, but reached significantly higher levels in the experimental groups than in the reference group. In adipose tissue, 22:6n-3 increased in the experimental groups during the study period, but decreased in the reference group, suggesting that a surplus of dietary 22:6n-3 was stored.

Reproducibility of automated periodontal probing around teeth and osseointegrated oral implants.

Three different probing devices (Audio-Probe, Florida-Probe, Peri-Probe) were tested in order to determine the clinical probing depth (CPD) around clinically stable oral implants and their homologous teeth and to evaluate their reproducibility. In all 37 patients, in the age range of 24-80 years, who had undergone periodontal therapy and placement of 1 or more oral implants (ITI), were selected for the study. The CPD was determined on 75 oral implants in total and at 4 sites of both the implants and the control teeth at 3 visits, each 1 week apart. At the 1st visit, the Florida-Probe and the Audio-Probe were used. At the 2nd visit, the Florida-Probe and the Peri-Probe and, at the 3rd visit, again, the Florida-Probe and the Audio-Probe were used. At each visit bleeding on probing (BOP) was registered. A statistically significant (P < 0.05) difference between the mean scores of implant and tooth sites was found showing slightly higher values for implant sites. A tendency for the deeper pockets to bleed more frequently than the shallow pockets was observed. The comparisons of differences of the readings of the Audio-Probe on 2 different occasions were smaller than for the Florida-Probe. However, comparisons between 2 different probes showed significantly greater measurement errors than when comparing the probes alone. There was a tendency for the Peri-Probe to yield the highest and the Audio-Probe the lowest values in inflamed sites. It was concluded that all 3 probing devices appeared to have adequate reproducibility both around teeth and oral implants. For clinical use in daily practice, the Audio-Probe was found to be the most simple device with the highest reproducibility.

[Tension-free inguinal and femoral hernioplasty using a mesh of plastic material]. / Tensionsfri ingvinal- og femoralhernioplastik med kunststofnet.

This study represents 202 consecutive cases of inguinal- or femoral hernias treated at the Department of Surgery, Silkeborg Central Hospital, Denmark. All were operated with the tension-free mesh-plug technique as described by Rutkow & Robbins (4). The median duration of the operation for the primary hernias was 35 (30-45) minutes, (median(25-75 percentile)), and 45 (30-60) minutes for the recurrent hernias. The operation was performed as a day-in procedure in 62% of the patients, whereas 35% stayed in the ward overnight. All daily activities, including strenuous exercises, were resumed within seven (2-14) days. Seven patients developed haematomas, none of which required treatment. One case of superficial infection required opening of the wound. Three recurrences have been reoperated, all using the mesh-plug technique and all with good results, while another three recurrences are suspected in patients with no or few complaints. It is concluded that inguinal- and femoral mesh-plug hernioplasty is a safe, efficient and low-cost procedure.

Influence of mercuric chloride on resistance to generalized infection with herpes simplex virus type 2 in mice.

The effect of mercuric chloride on resistance to generalized infection with herpes simplex virus type 2 (HSV-2) in mice was studied. The severity of the infection was evaluated by the amount of infectious virus in the liver. Mercury at a single dose of 20 micrograms aggravated the infection, and neither increasing the single dose to 80 micrograms nor giving repeated doses of 20 micrograms further intensified the infection. Examination of the course of infection after mercury exposure revealed an increased virus replication and dissemination during the first days of the infection, indicating that the early, nonspecific defence mechanisms were affected. Virus clearance and elimination, which is mediated by specific immunity, seemed not to be influenced. Examination of cells from the peritoneal cavity and of livers from virus-infected mice showed that mercury detectable by autometallography was exclusively found in mature peritoneal macrophages and in Kupffer cells of the liver. Inflammatory cells, recruited to the peritoneal cavity or infiltrating the infectious foci of the liver, did not show any mercury deposits. Attempts to demonstrate an effect in vivo of mercury on potential antiviral macrophage functions like interferon-alpha/beta (IFN-alpha/beta) and tumour necrosis factor-alpha (TNF-alpha) secretion and oxidative burst capacity were not successful, possibly because recruited, inflammatory cells, which have not been exposed to the high mercury concentrations at the site of injection, take over these functions of intoxicated macrophages.

Histochemical localization of autometallographically detectable mercury in tissues of the immune system from mice exposed to mercuric chloride.

The distribution of mercury in the spleen, liver, lymph nodes, thymus and bone marrow was studied by autometallography in mice exposed to mercuric chloride intraperitoneally. Application of immunofluorescence histochemistry and an autometallographic silver amplification method was employed to the same tissue section. Mercury was not only detected in macrophages marked by the antibody M1/70 but also in macrophage-like cells, which were either autofluorescent or devoid of fluorescent signals. These two cell types were identified as macrophages at the electron microscopical level. Autometallographically stained macrophages were observed in the spleen, lymph nodes, thymus and in Kupffer cells of the liver. Furthermore, mercury was observed in endothelial cells. No obvious pathological disturbances were observed at light and electron microscopical level. At the subcellular level mercury was localized in lysosomes of macrophages and endothelial cells.

Effect of mercuric chloride on macrophage-mediated resistance mechanisms against infection with herpes simplex virus type 2.

Macrophages play an important role in the early, nonspecific resistance to infection with herpes simplex virus. Mercuric chloride (HgCl2) accumulates in macrophages and has in certain concentrations a marked influence on the functional capacity of these cells. Therefore the influence of HgCl2 on resistance to generalized infection with herpes simplex virus type 2 (HSV-2) in mice and its effect on the HSV-2-induced activation of macrophages in vitro was examined. Mice injected intraperitoneally with HgCl2 24 h before infection with HSV-2 had more than 100 times higher virus titres in the liver 4 days after infection than mice not receiving any mercury. HgCl2 exerted a toxic effect on macrophages in vitro, which was especially pronounced during their adherence. Macrophages infected with HSV-2 were activated for an enhanced respiratory burst. This activation was abolished by treatment of the cells for 24 h with relatively low concentrations of HgCl2, resulting in macrophages with a potential to react with a respiratory burst comparable to that of uninfected cells. The HSV-2-induced activation of macrophages is mediated through the production and synergistic interaction of interferon-alpha/beta and tumour necrosis factor-alpha in an autocrine manner. The ability of these cytokines to activate macrophages and to interact synergistically was not affected by mercury. However the production by macrophages of both cytokines during the HSV-2 infection, but especially interferon-alpha/beta, which is essential for the activation, was reduced at low concentrations of HgCl2. Collectively these data indicate that mercury, by interfering with the early macrophage-production of cytokines, disables the early control of virus replication, leading to an enhanced infection.

29-week doxazosin treatment in patients with symptomatic benign prostatic hyperplasia. A double-blind placebo-controlled study.

In a placebo-controlled study, the safety and efficacy of the selective alpha 1-adrenoceptor-blocking agent doxazosin 4 mg once daily in the symptomatic treatment of benign prostatic hyperplasia (BPH) were evaluated. One hundred patients were primarily included in a 9-weeks study, and after this 75 patients accepted to continue in the present 20 weeks extension. Of the patients in the doxazosin-group (DG) 61% reported overall improvement against 53% in the placebo-group (PG)--(p = 0.56). In the DG, 49% of obstructive symptoms were improved compared to 27% in the PG (p < 0.01), and a reduction of 60% of irritative symptoms was found in the DG against 36% in the PG (p < 0.01). Daytime frequency was reduced by median 1.5 in the DG and remained unchanged in the PG (p < 0.01). Nocturia was reduced by median 1 and 0.5 respectively (p = 0.06). Maximum urinary flow rate (MFR) was improved by median 1.5 ml/s in the DG, while it deteriorated by median 0.5 ml/s in the PG (p < 0.05), Considering postvoid residual urine volume, cystometry variables (first sensation and bladder capacity), changes in sexual function and adverse events there was no difference between the two groups. In conclusion, doxazosin 4 mg once daily in long-term treatment of patients with BPH reduces both obstructive and irritative symptoms, daytime voiding frequency and although only slightly, significantly augments MFR without interference with sexual function and without other serious adverse effects.

Methyl mercury, mercuric chloride, and silver lactate decrease superoxide anion formation and chemotaxis in human polymorphonuclear leucocytes.

Cytotoxic effects of mercuric chloride, methyl mercury, and silver lactate on polymorphonuclear leucocytes have been examined by assaying superoxide anion formation capability and chemotaxis of metal-exposed cells. Both superoxide anion formation and chemotaxis were negatively affected by all three metal compounds. Both bacteriotoxic functions were affected in a dose-dependent fashion, the functional deficits were seen at doses not affecting cell viability. Dose-response curves were remarkably similar for all three compounds. The bacteriotoxic capacity of polymorphonuclear leucocytes may be hampered by mercury and silver.

Differentiation of silver-enhanced mercury and gold in tissue sections of rat dorsal root ganglia.

Autometallography was used in conjunction with light and electron microscopy to detect traces of gold and mercury in the dorsal root ganglia of rats treated with sodium aurothiomalate and mercuric chloride. In order to differentiate between gold and mercury in tissue sections, the gold accumulations were removed by potassium cyanide, leaving mercury sulphides/selenides as the only possible catalysts for autometallographic development. With this technique, it is now possible to differentiate between all tissue metals capable of initiating the autometallographic process, i.e. gold, vesicular zinc, and sulphides and selenides of mercury and silver.

Doxazosin treatment in patients with prostatic obstruction. A double-blind placebo-controlled study.

The safety and efficacy of the selective alpha 1-blocking agent doxazosin 4 mg once daily in the symptomatic treatment of benign prostatic hyperplasia were evaluated in a randomized, double-blind and placebo-controlled 9-week study of 100 patients. By patients' overall assessment of voiding difficulties, 79% in the doxazosin group (DG) and 44% in the placebo group (PG) reported improvement (p = 0.001). In the DG, improvement was noted in 63% of obstructive symptoms compared to 32% in the PG (p = 0.015), whereas improvement was noted in 76% and 45%, respectively, of irritative symptoms (p = 0.12). Daytime frequency was reduced by 1.5 in the DG and increased by 0.3 in the PG (p = 0.001), and nocturia was reduced by 1.1 and 1.0, respectively (p = 0.12). Maximum urinary flow rate was improved by 1.5 ml/s in the DG, while it deteriorated by 0.3 ml/s in the PG (p = 0.11). Considering postvoid residual urine volume, cystometry variables (first sensation and bladder capacity) and adverse events there was no difference between the two groups. In conclusion, doxazosin 4 mg once daily is safe and effective in relieving symptoms in patients with BPH.

Comparison of the interaction of methyl mercury and mercuric chloride with murine macrophages.

The toxicity of organic methyl mercury was studied on murine macrophages in cell culture and compared to that of inorganic mercuric chloride. Long-term treatment of macrophage cultures with methyl mercury resulted in decreased cell viability in a concentration-dependent fashion. Experiments showed that 20 microM methyl mercury was highly toxic, causing cell death within a few days, while cultures exposed to lower levels were less severely affected. Comparison of the toxicity of organic and inorganic mercury by cell viability showed no difference between equimolar concentrations of methyl mercury and mercuric chloride. Furthermore, protein synthesis (interferon-alpha/beta) was reduced in a concentration dependent manner and had the same reduced magnitude in cells treated with either methyl mercury or mercuric chloride. However, impairment of random migration and phagocytosis of macrophages appeared at lower concentrations in cells exposed to methyl mercury than in cells exposed to mercuric chloride. Electron microscopy of cells exposed to methyl mercury revealed mercury deposits in lysosomes and dispersed in the cytoplasm and nuclei. The present study shows that methyl mercury and mercuric chloride impair cell viability and protein production in cell cultures at equimolar concentrations, while methyl mercury inhibits macrophage functions such as migration and phagocytosis at lower concentrations than mercuric chloride.

In vitro effects of bladder mucosa and an enkephalinase inhibitor on tachykinin induced contractility of the dog bladder.

Tachykinin-induced contractility of smooth muscle strips from dog bladders was studied in vitro, and the presence of substance P-like immunoreactivity and neurokinin A and neurokinin B-like immunoreactivity was examined in bladder sections. Nerve fibers with substance P-like immunoreactivity were present in the mucosa, submucosa and smooth muscle. Fibers were also found in nerves, intramural ganglia, and around blood vessels. Neurokinin A-like immunoreactivity had similar distribution, and no neurokinin B-like immunoreactivity was observed. Removal of the mucosa significantly enhanced the sensitivity and the maximum responses to the tachykinins. After removing the mucosa, the sensitivity to these tachykinins increased 0.4 to 0.5 log units (p less than 0.02). The responses to carbachol were not altered by mucosa removal. The leftward shifts of the concentration-response curves for neurokinin A were of similar magnitude after removal of the mucosa, and after pretreatment with phosphoramidon (10 microM), an enkephalinase inhibitor, in the presence of mucosa. However, phosphoramidon did not alter the sensitivity of the bladder strips to neurokinin B, and slightly changed the sensitivity to substance P (0.2 log units). Additional shifts of the substance P and neurokinin A curves to the left were observed in the presence of phosphoramidon when the mucosa was removed (0.6 and 0.5 log units, p less than 0.005). The order of potency for the tachykinins (neurokinin A greater than substance P) was not altered by mucosa removal, addition of phosphoramidon, or both. Neurokinin A was degraded by enkephalinase located in the bladder mucosa and addition of phosphoramidon or mucosa removal resulted in an inhibition or loss of enkephalinase activity. It is concluded that the responses to neurokinin A, which acts on NK-2 type of receptors, prevail on the dog bladder.