Control Strips Research Articles

Reliable urine analysis in the management of hypertensive pregnancies

Objective: To determine the intra- and inter-observer precision of the Clinitek ® 50 automated urinalysis system. Setting: A teaching hospital. Methods: Urine samples were tested by seven operators with Multistix 8SG reagent strips, in duplicate and in replicates of five by the same operators, and by groups of four operators on the same samples. Precision was assessed using true urine samples and Chek-Stix control strips. Results: All 32 samples tested in duplicate and in replicates of five agreed to within one colour block by between 99.4 and 100% for all eight analytes. Seven samples tested by four different operators also gave 100% agreement to within one colour block for nearly all analytes. All operators found the Clinitek 50 system to be quick and easy to use. Conclusion: The Clinitek ® 50 automated urinalysis device demonstrates high intra- and inter-observer precision for duplicate, multiple replicate and multiuser testing with consistent performance. There is potential to improve antenatal care by introducing this technology into hospital or home-based care.

UP-REGULATION OF BRADYKININ RESPONSE IN RAT AND HUMAN BLADDER SMOOTH MUSCLE

Responses to bradykinin were investigated in vitro in isolated control and hypertrophic smooth muscle strips from rat bladder. Bladder hypertrophy was induced by a 10-day period of partial urinary outflow obstruction. In addition, human bladder strips were also investigated. Bradykinin (1 nM. to 1 microM.) caused contractions in all tissues studied. In the freshly isolated rat bladder preparations bradykinin induced contractions were similar and of small amplitude in control and hypertrophic tissues. After a 4-hour equilibratory period contractile responses to bradykinin and the B1 specific bradykinin receptor agonist desArg9 bradykinin were slightly increased in the controls but there was approximately a 6-fold increase in the hypertrophic muscle strips. After 4 hours of equilibration the human bladder strips showed a smaller but still significant increase in contractile response to bradykinin. Indomethacin, a cyclooxygenase inhibitor, almost abolished the increased response, which suggests that prostanoids are involved in the up-regulated response. The protein synthesis inhibitor cycloheximide inhibited up-regulation by approximately 50% in hypertrophic and control muscle strips from rat bladder and normal muscle from human bladder. These results demonstrate that bradykinin receptor responses are present in rat and human detrusor muscle and they can be up-regulated in vitro. Experiments on hypertrophic rat bladder revealed that this process is enhanced in hypertrophy.

Co-incubation of native and oxidized low-density lipoproteins: potentiation of relaxation impairment

The influence of native low-density lipoprotein (LDL) on the inhibition of endothelium-dependent relaxation previously induced by oxidized LDL was investigated with intact rabbit aortic rings. We also tried to assess oxysterol involvement in the native lipoprotein effects. Lipoprotein fractions (1 mg protein/ml) were tested for their ability to inhibit the vasorelaxation induced by acetylcholine in aorta rings previously precontracted by noradrenaline vs. that in control strips in Krebs buffer. Co-incubation of oxidized and native LDL reinforced the oxidized LDL-induced inhibition, compared to the impairment evoked by oxidized LDL alone ( E max=43.3±6.7% and 61.4±5.4%, respectively; P<0.05). Finally, smaller amounts of 7-oxy-cholesterols were recovered in organ baths after co-incubation of native and oxidized LDL than after incubation of oxidized LDL alone. Conversely, more oxy-c1holesterols were found in the strip vessels under the same conditions (% of oxysterol incorporation: 0.05158 vs. 0.10199, r=0.703). Together these results suggest that the strengthening of oxidized LDL-induced inhibition by native LDL is dependent on an oxysterol effect on arterial wall cells. Mechanisms involved in this phenomenon remain to be investigated.

Regeneration of White Spruce Under Aspen Canopies: Seeding, Planting, and Site Preparation

Abstract This study investigated the establishment and survival of naturally and artificially seeded white spruce (Picea glauca), as well as three sizes of planted white spruce stock, on different types of scarification beneath mature aspen (Populus tremuloides) in northern Alberta. White spruce seed rain, natural and hand-seeded regeneration establishment and survival, and understory vegetation response were monitored for 4 yr on scarified strips (light blading, heavy blading, and ridged) and on undisturbed controls. Despite a heavy seed rain in 1993, seedling establishment on control plots was almost nil. On scarified plots, the number of seeds required to produce a live seedling after 3 growing seasons ranged from 15 to 37 and up to 68 in one site. Most mortality occurred during the summer. Germination rate on the hand-seeded plots ranged from 19 to 28% on the scarified strips. There was no significant difference in survival from seed (15%) after three growing seasons among the three scarification treatments. Survival of planted stock was 98% on scarified strips and 96% on control strips. Diameter growth was least on control plots but was not significantly different among the scarified treatments. In contrast, height increment was greatest on the light blading treatment and differed little among the other two treatments and control. These results suggest that spruce regeneration in aspen forests can be promoted by scarification or underplanting. West. J. Appl. For. 15(4):177–182.

Abnormal contractile properties of rectal smooth muscle in chronic ulcerative colitis.

Patients with ulcerative colitis have abnormal rectal motility. To compare the contractile properties of rectal smooth muscle from patients with ulcerative colitis and controls. Rectal smooth muscle strips from patients undergoing resection for ulcerative colitis or cancer (control) were mounted in an organ bath. The effects of carbachol (receptor-mediated) and potassium (causes membrane depolarization) were studied. Acetylcholinesterase histochemistry was performed and nerve counts compared. Ulcerative colitis (n=41) and control (n=34) strips contracted in response to potassium and carbachol. Mean (S.E. M.) maximum response to potassium in the control and ulcerative colitis groups was 1.07 (0.06) g/mg and 1.02 (0.09) g/mg tissue, respectively (P=N.S.). EC50s (concentrations required to give 50% of maximal response) were 75 (1) mM and 73 (1) mM, respectively (P=N.S. ). Although maximum responses to carbachol were similar, 2.12 (0.12) g/mg and 1.95 (0.12) g/mg tissue (P=N.S.), ulcerative colitis strips exhibited an increased sensitivity to carbachol, EC50s: 5.05 x 10-6 (0.55 x 10-6) M vs. 8.36 x 10-6 (0.88 x 10-6) M, P=0.002). There was no significant difference in nerve counts between the tissues, as assessed by staining for acetylcholinesterase. Ulcerative colitis tissue has an increased sensitivity to carbachol and this is not due to denervation; it may result from increased calcium release from intracellular stores since contraction due to membrane depolarization is not altered. Modulation of this pathway could potentially be used to alter rectal motility in patients with ulcerative colitis.

Effect of serum concentration on Candida biofilm formation on acrylic surfaces.

The biofilm formation of the oral fungal pathogen Candida on denture acrylic strips coated with saliva, serum and, saliva-serum pellicle were examined in vitro using Candida albicans (four isolates), Candida glabrata (three isolates) and Candida tropicalis (three isolates). The degree of biofilm activity varied depending upon both the isolate and the pellicle. Significantly increased biofilm activity on the pellicle (particularly serum)-coated strips was observed with three isolates of C. albicans and another of C. glabrata on protein-coated acrylics, with increasing concentration of serum in the pellicle. Similar trends were observed with one isolate of C. albicans and C. glabrata, although the effects of pellicles were not significant. In contrast, with all three isolates of C. tropicalis and a single isolate of C. glabrata, although the biofilm activity on the protein-free control strips was significantly higher than that of saliva-coated strips, the increase in activity of pellicle-admixed biofilm depended upon the serum concentration. Candidal biofilm formation on acrylic surfaces is essentially promoted with increasing concentration of serum in the pellicle. This suggests that inflammation in the oral environment would facilitate fungal colonization on denture acrylic.

S-nitrosoglutathione-induced decrease in calcium sensitivity of airway smooth muscle.

The purpose of this study was to examine whether the nitric oxide donor S-nitrosoglutathione (GSNO) relaxes canine tracheal smooth muscle (CTSM) strips by decreasing Ca(2+) sensitivity [i.e., the amount of force for a given intracellular Ca(2+) concentration ([Ca(2+)](i))]. We further investigated whether GSNO decreases Ca(2+) sensitivity by altering the relationship between regulatory myosin light chain (rMLC) phosphorylation and [Ca(2+)](i) and the relationship between force and rMLC phosphorylation. GSNO (100 microM) relaxed intact CTSM strips contracted with 45 mM KCl by decreasing Ca(2+) sensitivity in comparison to control strips without significantly decreasing [Ca(2+)](i). GSNO reduced the amount of rMLC phosphorylation for a given [Ca(2+)](i) but did not affect the relationship between isometric force and rMLC phosphorylation. These results show that in CTSM strips contracted with KCl, GSNO decreases Ca(2+) sensitivity by affecting the level of rMLC phosphorylation for a given [Ca(2+)](i), suggesting that myosin light chain kinase is inhibited or that smooth muscle protein phosphatases are activated by GSNO.

Efeitos da faixa de controle do capim-braquiária (Brachiaria decumbens) no desenvolvimento inicial de plantas de eucalipto

Esta pesquisa foi conduzida em área reflorestada com Eucalyptus grandis x Eucalyptus urophylla, no município de Três Lagoas, MS, no período de julho de 1996 a agosto de 1997, com o objetivo de avaliar os efeitos da variação da faixa de controle de Brachiaria decumbens ao longo da linha de plantio sobre o desenvolvimento inicial de clones de eucalipto. Os tratamentos experimentais constaram de dois grupos: no primeiro foram mantidas faixas constantes de controle durante os 12 meses iniciais após o transplante das mudas no campo: 0, 25, 50, 100, 125 e 150 cm de cada lado da linha de transplante das mudas de eucalipto; e, no segundo, foram adotadas faixas crescentes de controle ao longo do período experimental. Ao final do período experimental (390 dias após o transplante), constatou-se que as plantas de eucalipto que cresceram nas parcelas com faixas de controle constantes ou crescentes, iguais ou superiores a 100 cm, mostraram-se superiores, em diâmetro, altura e velocidade de crescimento absoluto. Com base nesses resultados, pode-se concluir que a largura mínima da faixa de controle a ser utilizada é de 100 cm de cada lado da linha, a fim de manter as plantas de eucalipto livres da interferência das plantas daninhas.

Selective Alkylation of Rat Urinary Bladder Muscarinic Receptors with 4-DAMP Mustard Reveals a Contractile Function for the M2 Muscarinic Receptor

Our previous data indicate that M3 muscarinic receptors mediate carbachol induced bladder contractions. The data presented here were obtained by selective alkylation of M3 receptors with 4-DAMP mustard and suggest that the M2 receptor subtype may be involved in inhibition of β-adrenergic receptor induced relaxation, therefore, allowing recontraction. Alkylation resulted in 85% of M3 receptors and 65% of M2 receptors unable to bind radioligand as demonstrated by subtype selective immunoprecipitation. Rat bladder strips subjected to our alkylation procedure contracted submaximally, and direct carbachol contractions were inhibited by antagonists with affinities consistent with M3 receptor mediated contraction. In contrast, the affinities of antagonists for inhibition of carbachol induced recontractions following isoproterenol stimulated relaxation in the presence of 90 mM KCI, indicated a contractile function for the M2 receptor that was not observed in control strips. In conclusion, these studies demonstrate a possible role for the M2 subtype in bladder smooth muscle contraction.

Sterically Inhomogenous Viscoelastic Behavior of Human Saccular Cerebral Aneurysms

To clarify the mechanism leading to the development and rupture of intracranial aneurysms, tensile strength and viscoelastic parameters of 22 human saccular aneurysms were investigated. Meridional and circumferential strips from the thin and the thick part of the aneurysm sack and 18 control strips from the basilar artery of 8 patients with pathologies not affecting the cerebral arterial system were studied. The length of the strips was increased in 200-µm steps, while distending force was recorded. Tensile strength and viscoelastic parameters were computed. In both directions, tensile strength of thick strips was significantly lower than that of controls. In the meridional direction, tensile strength of thin strips was significantly larger than that of thick ones (14.5 ± 4.1 × 10⁶ vs. 7.5 ± 2.0 × 10⁶ dyn/cm², p < 0.05). In the circumferential direction, thin strips tore at lower strain values than thick ones (29 ± 4 vs. 55 ± 16%, p < 0.05). Viscoelastic parameters changed in parallel. In circumferential direction, values of thick and thin strips were significantly lower than those of controls. In the meridional direction, values of thin strips were significantly higher than those of the thick ones. These observations show that characteristic mechanical deterioration and steric inhomogeneities accompany the loss of smooth muscle cells and the derangement of connective tissue elements in the wall of intracranial aneurysms, which may explain certain steps in their initiation, enlargement and rupture.

EFFECT OF ADDITIONAL TESTOSTERONE ON PURINERGIC RESPONSES IN ISOLATED RABBIT CORPUS CAVERNOSUM STRIPS

Recently, it has been demonstrated that the endothelium of corpus cavernosum (CC) plays an important role on smooth muscle relaxation, which is crucial to initiate and maintain erection. We investigated the effect of long-lasting additional testosterone propionate (TP) therapy on endothelium-dependent and -independent relaxations of isolated rabbit CC. Isolated CC strips were mounted in organ baths and isometric tension was recorded. Addition of a specific inhibitor of nitric oxide synthesis,NG-nitro-L-arginin methyl ester (l-NAME), into the organ bath had no effect on the relaxation responses to adenosine (ADO), adenosine 5′-triphosphate (ATP) and sodium nitroprusside (SNP) in isolated CC strips precontracted with phenylepherine, but completely inhibited relaxation responses produced by ADP. Adenosine and adenine nucleotides relaxed the phenylepherine-induced contractile response in control strips with the potency order: ADO (62.8±3.2%) > ATP (37.1±5.2%) > ADP (25.8±2.5%). The relaxation responses to ADO, ATP and SNP in isolated rabbit CC strips were not significantly altered by additional TP therapy. The relaxation responses produced by ADP were significantly enhanced following 1 and 2 months TP therapy as compared with controls. However, in the group treated with TP for 2 months followed by a 2 months drug-free period, relaxation responses were significantly reduced compared to 1 and 2 months treatment groups, and approached control values. Increased relaxation responses to ADP following 1 and 2 months additional TP therapy may be a result of increased endothelial purinergic receptor density, or it may be due to stimulation of the synthesis and/or release of endothelial nitric oxide (NO) by TP.

Clostridium difficile toxin B inhibits carbachol‐induced force and myosin light chain phosphorylation in guinea‐pig smooth muscle: role of Rho proteins

1. Clostridium difficile toxin B glucosylates the Ras-related low molecular mass GTPases of the Rho subfamily thereby inactivating them. In the present report, toxin B was applied as a tool to test whether Rho proteins participate in the carbachol-induced increase in the Ca2+ sensitivity of force and myosin light chain (MLC) phosphorylation in intact intestinal smooth muscle. 2. Small strips of the longitudinal muscle of guinea-pig small intestine were incubated in toxin B (40 ng ml-1) overnight. Carbachol-induced force and intracellular [Ca2+], and, in a separate series, force and MLC phosphorylation, were determined. 3. Carbachol induced a biphasic contraction: an initial rapid increase in force (peak 1) followed by a partial relaxation and a second delayed increase in force (peak 2). The peak of the Ca2+ signal measured with fura-2 preceded peak 1 of force and then declined to a lower suprabasal steady-state level. Peak 2 was not associated with a significant increase in [Ca2+]. Toxin B nearly completely inhibited peak 2 while peak 1 was not significantly inhibited. Toxin B had no effect on the Ca2+ transient. 4. In control strips, MLC phosphorylation at peak 2 was 27.7% which was significantly higher than the resting value (18.6%). The inhibition of the second, delayed, rise in force induced by toxin B was associated with complete inhibition of the increase in MLC phosphorylation. The resting MLC phosphorylation was not significantly different from that of the control strips. 5. The initial increase in MLC phosphorylation determined 3 s after exposure to carbachol was 54% in the control strips. Toxin B also inhibited this initial phosphorylation peak despite the fact that the Ca2+ transient and the initial increase in force were not inhibited by toxin B. This suggests that Rho proteins play an important role in setting the balance between MLC phosphorylation and dephosphorylation reactions even at high levels of intracellular Ca2+. 6. These findings are consistent with the hypothesis that the delayed rise in force elicited by carbachol is due to an increase in the Ca2+ sensitivity of MLC phosphorylation mediated by Rho proteins.

Forestry Practices and the Risk of Bird Nest Predation in a Boreal Coniferous Forest

Reconciling tree harvesting with the maintenance of forest bird populations is a major concern of integrated management. Because bird nest predation causes >50% of the nest losses in passerines and is known to vary according to habitat characteristics, we explored some aspects of avian nest predation in relation to forestry practices in a boreal coniferous landscape managed primarily for timber production in Québec. Using artificial tree and ground nests with Common Quail (Coturnix coturnix) and plasticine eggs, we compared the risk of nest predation (1) in experimental riparian forest strips of different widths (20-m, 40-m, and 60-m unthinned strips; >300-m control strips; and 20-m thinned strips) and (2) in clearcuts experimentally subjected to different regeneration practices (plantations with chemical and mechanical weeding, and naturally regenerated clearcuts) between 1992 and 1995. The risk of ground nest predation was lower in naturally regenerated clearcuts (5% daily probability of predation) than in control forest strips (27%). We found no evidence that chemical and mechanical weeding affected the risk of nest predation in clearcuts. In forest strips, the predation risk was higher in forest strips 40–60 m wide than in 20-m and control strips. Birds accounted for 13% of predation signs, whereas red squirrels (Tamiasciurus hudsonicus) were the dominant mammalian predators, accounting for 36% of the total predation (n = 201 nests preyed upon). In our region, the low predation rates (30% for 24 real nests) and the absence of generalist foragers such as crows, raccoons, and skunks could be attributed to the near absence of human occupation in comparison to forest-dominated landscapes in Europe and northeastern United States.

FORESTRY PRACTICES AND THE RISK OF BIRD NEST PREDATION IN A BOREAL CONIFEROUS FOREST

Reconciling tree harvesting with the maintenance of forest bird populations is a major concern of integrated management. Because bird nest predation causes >50% of the nest losses in passerines and is known to vary according to habitat characteristics, we explored some aspects of avian nest predation in relation to forestry practices in a boreal coniferous landscape managed primarily for timber production in Quebec. Using artificial tree and ground nests with Common Quail (Coturnix coturnix) and plasticine eggs, we compared the risk of nest predation (1) in experimental riparian forest strips of different widths (20-m, 40-m, and 60-m unthinned strips; >300-m control strips; and 20-m thinned strips) and (2) in clearcuts experimentally subjected to different regeneration practices (plantations with chemical and mechanical weeding, and naturally regenerated clearcuts) between 1992 and 1995. The risk of ground nest predation was lower in naturally regenerated clearcuts (5% daily probability of predation) than...

The relaxant effect of ketamine on guinea pig airway smooth muscle is epithelium-independent.

Airway epithelial cells and vascular endothelial cells modulate the tone of the underlying smooth muscle by releasing relaxing factors such as prostanoids and nitric oxide (NO). In the present study, we investigated whether the relaxant effect of ketamine depends on any of the epithelium-derived relaxing factors. Tracheae of female guinea pigs were cut spirally into strips (15 x 3 mm) and mounted in water-jacketed organ baths filled with Krebs-bicarbonate buffer aerated with a mixture of 95% O2 and 5% CO2 at 37 degrees C. Changes in the tension of the strips were measured isometrically with a force displacement transducer and recorded with a polygraph. In the first set of experiments, we examined the effect of ketamine on the concentration-response curves for histamine and carbachol in strips in which the epithelium was kept intact and in strips with denuded epithelium. In the second and third set of experiments, we studied the effect of indomethacin, a cyclooxygenase inhibitor, and N-omega-nitro-L-arginine methylester(L-NAME), a NO synthase inhibitor, on the relaxant activity of ketamine on tracheal strips contracted by histamine or carbachol. The following results were obtained: 1. Mechanical denudation of the tracheal epithelium shifted the concentration-response curve for histamine to the left (the 50% effective concentration [EC50] value of histamine decreased from 3.5 +/- 0.02 x 10(-6) M in the intact strips to 0.98 +/- 0.01 x 10(-6) M in denuded strips, P < 0.001). However, removal of the tracheal epithelium did not change the response to carbachol (the EC50 for carbachol was 1.1 +/- 0.02 x 10(-7) M in intact strips versus 0.88 +/- 0.01 x 10(-7) M after epithelial removal, P > 0.05). 2. Ketamine shifted to the right the concentration-response curves for histamine and carbachol in both intact and denuded tracheae. 3. Indomethacin did not alter the relaxant effect of ketamine on the tracheae contracted by either histamine (the concentration that inhibits 50% [IC50] of ketamine = 1.5 +/- 0.01 x 10(-3) M in control strips and 1.3 +/- 0.04 x 10(-3) M in strips pretreated with indomethacin, P > 0.05) or carbachol (the IC50 of ketamine was 2.5 +/- 0.02 x 10(-4) M in control strips and 2.4 +/- 0.01 x 10(-4) M in strips pretreated with indomethacin, P > 0.05). 4. L-NAME did not influence the relaxant effect of ketamine on tracheae contracted by either histamine (the IC50 of ketamine = 1.6 +/- 0.05 x 10(-3) M in control strips and 1.6 +/- 0.05 x 10(-3) M in strips pretreated with L-NAME, P > 0.05) or carbachol (the IC50 of ketamine = 2.6 +/- 0.04 x 10(-4) M in control strips and 2.3 +/- 0.01 x 10(-4) M in trips pretreated with L-NAME, P > 0.05). These results indicate that neither the mechanical removal of the tracheal epithelium nor the blockade of the release of potent mediators from tracheal epithelial cells influence the relaxant effect of ketamine on guinea pig tracheal strips contracted by histamine or carbachol. We conclude that ketamine relaxes the airway smooth muscle by an epithelium-independent mechanism.

Is guanosine-5'-triphosphate involved in calcium-activation of contractile proteins in vascular smooth muscle?

Isometric tension was measured to investigate the effects of guanosine-5'-triphosphate (GTP) on the run-down of myofilament Ca2+ sensitivity in isolated rat mesenteric arteries permeabilized with beta-escin. The Ca2+ sensitivity assessed by the EC50 value for the Ca2+ (0.1-100 microM)-tension relationship progressively runs down in the control strips, while it was well-preserved for 5-successive Ca2+ applications in the presence of GTP (50 microM); no significant difference was found in the Ca2+ sensitivity observed with the 1st Ca2+ application between the control and GTP-treated strips. Guanosine-5'-(2-O-thio) diphosphate (GDP beta S, 100 microM) significantly decreased the Ca2+ sensitivity with the 1st Ca2+ application and eliminated the run-down of Ca2+ sensitivity. GTP (3-150 microM), applied to the strips submaximally precontracted with Ca2+, had a little effect on the Ca2+ contractions in the early stage of experiments, but dramatically enhanced the Ca2+ contractions in their later stage; its latter effect was mimicked by guanosine-5'-(3-O-thio) triphosphate (GTP gamma S) and reversed by GDP beta S (100 microM). The results suggest: 1) loss of endogenous GTP following permeabilization is involved in the run-down of Ca2+ sensitivity; and 2) activation of G-proteins is involved in Ca(2+)-activation of contractile proteins.

Effects of dietary sugars and, saliva and serum on Candida bioflim formation on acrylic surfaces.

The effect of two dietary sugars, glucose and galactose, on biofilm formation of the oral fungal pathogen Candida on denture acrylic strips coated with saliva and serum pellicles was examined in vitro using Candida albicans (3 isolates), C. glabrata (2 isolates) and C. tropicalis (2 isolates). The degree of biofilm activity was affected by both the dietary sugar and the nature of the pellicle (ANOVA, p < 0.01). With most isolates the glucose grown yeasts demonstrated significantly more bioflim activity than the galactose grown fungi, in the presence of pellicles (ANOVA, p < 0.01 or P < 0.01). In contrast, one isolate of galactose-grown yeast elicited significantly higher biofilm activity than glucose-grown yeasts on the control strips (ANOVA, p < 0.01). Taken together, these results imply that a saliva or a serum pellicle, and the carbon source in the environment, act a complex manner modulating Candida bioflim formation.

Fate of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella typhimurium during Preparation and Storage of Beef Jerky

The fate of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella typhimurium during preparation and storage of beef jerky was determined. Control strips and one-half of the inoculated beef loin strips were marinated at 4°C overnight and dried at 60°C (140°F) for 10h. The remaining half of the inoculated samples were heated in marinade to 71.1°C (160°F). Strips were dried at 60°C (140°F) for 10 h. Microbial populations were determined at intervals during drying up to 10 h and also from samples stored at 25°C for 8 weeks at various moisture levels. In general, L. monocytogenes was more resistant to the treatments. After 3 h of drying, populations on the unheated, inoculated samples were reduced by 3.3, 1.8 and 3.1 log units, respectively, and all three were reduced by 5.5 to 6.0 log units after 10h. Reduction of the three populations on strips that were cooked prior to drying was 4.5 to 5.5 log units immediately after cooking. The populations decreased to undetectable levels after 10 h of drying. None of the three pathogens were detected on the controls. After 8 weeks of storage none of the pathogens were detected, indicating that they were unable to recover under the moisture conditions during storage.

A pharmacological and histochemical study of hamster urethra and the role of urothelium.

1. Electrical field stimulation (EFS) of circular strips of hamster proximal urethra caused frequency-dependent relaxations at raised tone. Phentolamine (10(-6) M), propranolol (10(-6) M) and atropine (10(-6) M) were present throughout the experiment. Neurogenic relaxation was attenuated by L-NG-nitroarginine methyl ester (L-NAME) (10(-4) M), was restored by L-arginine (3 x 10(-3) M) but not by D-arginine (3 x 10(-3) M) and completely blocked by tetrodotoxin (10(-6) M). Neurogenic relaxation was also reduced by suramin (10(-4) M) and totally blocked by suramin together with L-NAME. Strips of hamster urethra devoid of urothelium showed little, if any, relaxant response to EFS. 2. An immunohistochemical study showed nitric oxide synthase-immunoreactive nerves in the smooth muscle layers and in the lamina propria, just beneath the urothelium, but no nitric oxide synthase (NOS) staining in the urothelial layer. 3. Noradrenaline elicited a significantly greater contraction in strips without urothelium than in control strips. L-NAME (10(-4) M) did not affect noradrenaline-induced contraction in both control and urothelium-free strips. The contractile response to acetylcholine was not dependent on the presence or absence of urothelium. Nevertheless the response induced by exogenous acetylcholine (10(-3) M) was increased by L-NAME (10(-4) M), both in intact and in urothelium-free strips. 4. Prostaglandin E2 (10(-8)-5 x 10(-6) M) and 2-methyl-thio-ATP (10(-9)-10(-5) M) relaxed proximal urethra. Suramin (10(-4) M) significantly inhibited the relaxation induced by 2-methyl-thio-ATP. The amplitude of these responses was not significantly different between intact and urothelium-free strips and was not blocked by L-NAME (10(-4) M). 5. These results suggest that nitric oxide (NO) is the principal transmitter involved in the non-adrenergic, non-cholinergic (NANC) relaxation of hamster proximal urethra possibly together with another inhibitory transmitter released from nerves. NO can be released from nerves located in the circular smooth muscle layer and in the lamina propria rather than in the urothelium. The reduced neurogenic relaxation in urothelium-free preparations suggests that a NO-dependent inhibitory factor is released from the urothelium. In addition, ATP and prostaglandin E2 may be involved, together with NO, in the urethra during micturition.

Role of altered responsiveness of hypertrophic smooth muscle in manometric abnormalities of the obstructed opossum oesophagus.

The movements of the obstructed oesophagus are abnormal, but whether this relates to the disease causing the obstruction, to the altered load conditions or to abnormal neuromuscular functions in hypertrophic smooth muscle is unclear. In an opossum model of chronic oesophageal obstruction, we compared the mechanical responsiveness of hypertrophic smooth muscle in vitro to in vivo manometric function. Related to their greater thickness, strips of hypertrophic muscle generated greater force in response to electrical stimulation and to stretch than control strips. Hypertrophic muscle often generated repetitive contractions; spread of contractions orad from the stimulus site was common in hypertrophic oesophageal bands. On manometry, the obstructed oesophagus generated abnormally high pressures proximally, and highly variable pressure amplitudes in the middle and distally; pressure waves often occurred simultaneously throughout the oesophagus, were repetitive or multi-peaked and led to a lasting rise of oesophageal pressure. Alterations in the intrinsic neuromuscular functions of hypertrophic smooth muscle including generation of greater force, repetitive or spontaneous contractions, and retrograde spread of contractions explain many, but not all, of the manometric abnormalities seen in the chronically obstructed oesophagus.