Impact of an auditory stimulus on baseline cortisol concentrations in clinically normal dogs.

Baseline cortisol concentrations are routinely used to screen dogs for hypoadrenocorticism (HOC); this diagnosis must then be confirmed with an ACTH stimulation test. A baseline cortisol concentration less than 55 nmol/L (2 µg/dL) is highly sensitive for HOC but lacks specificity, with a false positive rate >20%. Many dogs with nonadrenal disease are therefore subjected to unnecessary additional testing. It was hypothesized that exposure to an unpleasant auditory stimulus before sample collection would improve the specificity of baseline cortisol measurements in dogs with nonadrenal disease by triggering cortisol production. Twenty-eight healthy client-owned dogs were included in the study, with a median age of 4 yr (range 2-9 yr) and a median weight of 20 kg (range 10-27 kg). Dogs were ineligible for inclusion if they had received short- or long-acting glucocorticoids within the previous 30 and 90 d, respectively. Dogs were randomly assigned to group 1 (control; no noise; n = 7), group 2 (brief noise: n = 10), or group 3 (long noise: n = 11). Each dog and owner were directed to a secluded area for approximately 15 min. Group 1 sat in relative quiet, exposed only to the background sounds of a veterinary hospital. Group 2 were exposed to the sound of a wet-dry vacuum in an adjacent hallway during the first 3 min of this period. Group 3 were exposed to random bursts of wet-dry vacuum noise during this period. At the end of the test interval, each dog was escorted to an adjacent examination room for blood collection. Samples were processed within 15 min; serum was frozen at -80°C before measurement of cortisol concentrations. Median serum cortisol concentrations and the proportion of dogs with results <55 nmol/L were similar for the 3 groups. The study hypothesis that exposure to the noise of a wet-dry vacuum cleaner would consistently drive baseline serum cortisol concentrations above 55 nmol/L in dogs with apparently normal adrenal function was therefore rejected.

Opportunistic Invasive Cutaneous Fungal Infections Associated with Administration of Cyclosporine to Dogs with Immune-mediated Disease.

BACKGROUND: Opportunistic invasive fungal infections (OIFIs) occur in dogs administered immunosuppressive medications. However, the epidemiology of OIFIs among dogs undergoing immunosuppressive treatment is poorly understood. The aims of this study were to (1) estimate the incidence of OIFIs among dogs diagnosed with certain immune-mediated diseases and treated with immunosuppressive drugs, and (2) determine if administration of particular drug(s) was a risk factor for OIFIs. HYPOTHESIS: Dogs receiving cyclosporine treatment (alone or as part of a multidrug protocol) are at higher risk of developing OIFIs. ANIMALS: One hundred and thirteen client-owned dogs diagnosed with select immune-mediated diseases: 42 with IMHA, 29 with ITP, 34 with IMPA, and 8 with Evans syndrome. METHODS: Retrospective cohort study. Medical records of dogs presenting to the Texas A&M University, Veterinary Medical Teaching Hospital between January 2008 and December 2015, and treated for 1 or more of IMHA, IMPA, ITP, or Evans syndrome were retrospectively reviewed. Dogs that did not develop an OIFI were excluded if they died, were euthanized, or were lost to follow-up within 120 days of initiation of immunosuppressive treatment. RESULTS: Fifteen dogs of 113 (13%) were diagnosed with an OIFI based on 1 or more of cytology, culture, or histopathology. The odds of developing an OIFI were greater among dogs that were treated with cyclosporine (OR = 7.1, P = 0.017; 95% CI, 1.5-34.4) and among male dogs (OR = 5.1, P = 0.018; 95% CI, 1.4-17.9). CONCLUSIONS AND CLINICAL IMPORTANCE: OIFIs were significantly more likely in male dogs and those receiving cyclosporine. It is important to consider OIFIs as a potential complication of immunosuppressive treatment, particularly cyclosporine.

Acute toll-like receptor 4 activation impairs rat renal microvascular autoregulatory behaviour.

AIM: Little is known about how toll-like receptor 4 (TLR4) influences the renal microvasculature. We hypothesized that acute TLR4 stimulation with lipopolysaccharide (LPS) impairs afferent arteriole autoregulatory behaviour, partially through reactive oxygen species (ROS). METHODS: We assessed afferent arteriole autoregulatory behaviour after LPS treatment (1 mg kg-1 ; i.p.) using the in vitro blood-perfused juxtamedullary nephron preparation. Autoregulatory behaviour was assessed by measuring diameter responses to stepwise changes in renal perfusion pressure. TLR4 expression was assessed by immunofluorescence, immunohistochemistry and Western blot analysis in the renal cortex and vasculature. RESULTS: Baseline arteriole diameter at 100 mmHg averaged 15.2 ± 1.2 µm and 12.2 ± 1.0 µm for control and LPS groups (P < 0.05) respectively. When perfusion pressure was increased in 15 mmHg increments from 65 to 170 mmHg, arteriole diameter in control kidneys decreased significantly to 69 ± 6% of baseline diameter. In the LPS-treated group, arteriole diameter remained essentially unchanged (103 ± 9% of baseline), indicating impaired autoregulatory behaviour. Pre-treatment with anti-TLR4 antibody or the TLR4 antagonist, LPS-RS, preserved autoregulatory behaviour during LPS treatment. P2 receptor reactivity was normal in control and LPS-treated rats. Pre-treatment with Losartan (angiotensin type 1 receptor blocker; (AT1 ) 2 mg kg-1 ; i.p.) increased baseline afferent arteriole diameter but did not preserve autoregulatory behaviour in LPS-treated rats. Acute exposure to Tempol (10-3 mol L-1 ), a superoxide dismutase mimetic, restored pressure-mediated vasoconstriction in kidneys from LPS-treated rats. CONCLUSION: These data demonstrate that TLR4 activation impairs afferent arteriole autoregulatory behaviour, partially through ROS, but independently of P2 and AT1 receptor activation.

[Bilateral renal sclerotherapy as a treatment option in a mixed breed male dog with idiopathic renal haematuria]. / Bilaterale Sklerotherapie zur Behandlung der idiopathischen renalen Hämaturie bei einem Mischlingsrüden.

A 2-year-old male castrated German Shepherd dog mix was presented with chronic macroscopic haematuria. Further diagnostics included abdominal ultrasound and urethrocystoscopy and led to a diagnosis of severe bilateral idiopathic renal haematuria (IRH). Medical treatment with Yunnan Baiyao was unsuccessful. Bilateral renal-sparing sclerotherapy was performed and, despite distal migration of both ureteral stents within 12 days, permanently resolved the macroscopic haematuria.

Evaluation of a catalase-based urine test for the detection of urinary tract infection in dogs and cats.

BACKGROUND: Bacterial infection of the urinary tract is a common disorder in dogs and cats. Although microscopic examination of urine sediment is routinely used to screen for infection, this test can lack sensitivity or require expertise. A reliable in-clinic screening test would be a useful adjunct for the identification of dogs and cats with bacterial urinary tract infection (UTI). HYPOTHESIS: That a catalase-based urine test (Accutest Uriscreen™) is a more sensitive screening test for UTI in dogs and cats than urine microscopic sediment examination. ANIMALS: One hundred and sixty client-owned dogs and cats. METHODS: Surplus urine from animals presented to a veterinary teaching hospital was used in this prospective observational study. A routine urinalysis, aerobic bacterial culture, and the Uriscreen test were performed on cystocentesis samples. Sensitivity and specificity with 95% confidence intervals and positive and negative likelihood ratios were calculated for Uriscreen and microscopic sediment examination using culture results as the gold standard. RESULTS: Bacterial culture was positive in 27/165 (16.4%) samples. The sensitivity, specificity, and positive and negative likelihood ratios for the Uriscreen were 89%, 71%, 3.0, and 0.15, respectively. Sensitivity, specificity, and positive and negative likelihood ratios for urine sediment microscopic examination were 78%, 90%, 7.8, and 0.24, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: The Uriscreen is a more sensitive screening test for UTI in dogs and cats than sediment examination; however, the urine sediment examination was more specific. A negative Uriscreen result helps exclude UTI; however, urine bacterial culture is still necessary to exclude or confirm UTI in all cases.

Sensitivity and specificity of a blood and urine galactomannan antigen assay for diagnosis of systemic aspergillosis in dogs.

BACKGROUND: Diagnosis of canine systemic aspergillosis requires fungal culture from a sterile site, or confirmatory histopathology from a nonsterile site. Invasive specimen collection techniques may be necessary. OBJECTIVE: To evaluate the sensitivity and specificity of a serum and urine Aspergillus galactomannan antigen (GMA) ELISA assay for diagnosis of systemic aspergillosis in dogs. DESIGN: Multicenter study. ANIMALS: Thirteen dogs with systemic aspergillosis and 89 dogs with other diseases. Thirty-seven of the 89 dogs had signs that resembled those of systemic aspergillosis and 52 dogs were not suspected to have aspergillosis. PROCEDURE: The GMA ELISA was performed on serum specimens from all dogs and urine specimens from 67 dogs. Galactomannan indices (GMI) ≥ 0.5 were considered positive. Results for dogs in each group were compared. RESULTS AND CONCLUSIONS: The sensitivity and specificity of the assay for serum were 92 and 86%, respectively, and for urine were 88 and 92%, respectively. False negatives were seen only in dogs with localized pulmonary aspergillosis. Use of a cutoff GMI of 1.5 increased specificity to 93% for both serum and urine without loss of sensitivity for diagnosis of disseminated infection. High-level false positives (> 1.5) occurred in dogs with other systemic mycoses and those treated with Plasmalyte. CLINICAL RELEVANCE: Serum and urine Aspergillus GMA ELISA is a noninvasive, sensitive, and specific test for the diagnosis of disseminated aspergillosis in dogs when a cutoff GMI of ≥ 1.5 is used.

The prevalence of hypocobalaminaemia in cats with spontaneous hyperthyroidism.

OBJECTIVES: To determine the prevalence of hypocobalaminaemia in cats with moderate to severe hyperthyroidism and to investigate the relationship between cobalamin status and selected haematologic parameters. METHODS: Serum cobalamin concentrations were measured in 76 spontaneously hyperthyroid cats [serum thyroxine (T(4) ) concentration ≥100 nmol/L] and 100 geriatric euthyroid cats. Erythrocyte and neutrophil counts in hyperthyroid cats with hypocobalaminaemia were compared with those in hyperthyroid cats with adequate serum cobalamin concentrations (≥290 ng/L). RESULTS: The median cobalamin concentration in hyperthyroid cats was lower than the control group (409 versus 672 ng/L; P=0·0040). In addition, 40·8% of hyperthyroid cats had subnormal serum cobalamin concentrations compared with 25% of controls (P=0·0336). Weak negative correlation (coefficient: -0·3281) was demonstrated between serum cobalamin and T(4) concentrations in the hyperthyroid population, and the median cobalamin concentration was lower in cats with T(4) above the median of 153 nmol/L compared with cats with T(4) below this value (P=0·0281). Hypocobalaminaemia was not associated with neutropenia or anaemia in hyperthyroid cats. CLINICAL SIGNIFICANCE: This study indicates that a substantial proportion of cats with T(4) ≥100 nmol/L are hypocobalaminaemic and suggests that hyperthyroidism directly or indirectly affects cobalamin uptake, excretion or utilisation in this species.

Renal autoregulation in P2X1 knockout mice.

Autoregulation of renal blood flow is an established physiological phenomenon, however the signalling mechanisms involved remain elusive. Autoregulatory adjustments in preglomerular resistance involve myogenic and tubuloglomerular feedback (TGF) influences. While there is general agreement on the participation of these two regulatory pathways, the signalling molecules and effector mechanisms have not been identified. Currently, there are two major hypotheses being considered to explain the mechanism by which TGF signals are transmitted from the macula densa to the afferent arteriole. The adenosine hypothesis proposes that the released adenosine triphosphate (ATP) is hydrolysed to adenosine and this product stimulates preglomerular vasoconstriction by activation of A(1) receptors on the afferent arteriole. Alternatively, the P2 receptor hypothesis postulates that ATP released from the macula densa directly stimulates afferent arteriolar vasoconstriction by activation of ATP-sensitive P2X(1) receptors. This hypothesis has emerged from the realization that P2X(1) receptors are heavily expressed along the preglomerular vasculature. Inactivation of P2X(1) receptors impairs autoregulatory responses while afferent arteriolar responses to A(1) adenosine receptor activation are retained. Autoregulatory behaviour is markedly attenuated in mice lacking P2X(1) receptors but responses to adenosine A(1) receptor activation remain intact. More recent experiments suggest that P2X(1) receptors play an essential role in TGF-dependent vasoconstriction of the afferent arteriole. Interruption of TGF-dependent influences on afferent arteriolar diameter, by papillectomy or furosemide treatment, significantly attenuated pressure-mediated afferent arteriolar vasoconstriction in wild-type mice but had no effect on the response in P2X(1) knockout mice. Collectively, these observations support an essential role for P2X(1) receptors in TGF-mediated afferent arteriolar vasoconstriction.

Dynamic interaction between myogenic and TGF mechanisms in afferent arteriolar blood flow autoregulation.

The dynamic activity of afferent arteriolar diameter (AAD) and blood flow (AABF) responses to a rapid step increase in renal arterial pressure (100-148 mmHg) was examined in the kidneys of normal Sprague-Dawley rats (n = 11) before [tubuloglomerular feedback (TGF)-intact] and after interruption of distal tubular flow (TGF-independent). Utilizing the in vitro blood-perfused juxtamedullary nephron preparation, fluctuations in AAD and erythrocyte velocity were sampled by using analog-to-digital computerized conversion, video microscopy, image shearing, and fast-frame, slow-frame techniques. These assessments enabled dynamic characterization of the autonomous actions and collective interactions between the myogenic and TGF mechanisms at the level of the afferent arteriole. The TGF-intact and TGF-independent systems exhibited common initial (0-24 vs. 0-13 s, respectively) response slope kinetics (-0.53 vs. -0.47% DeltaAAD/s; respectively) yet different maximum vasoconstrictive magnitude (-11.28 +/- 0.1 vs. -7. 02 +/- 0.9% DeltaAAD; P < 0.05, respectively). The initial AABF responses similarly exhibited similar kinetics but differing magnitudes. In contrast, during the sustained pressure input (13-97 s), the maximum vasoconstrictor magnitude (-7.02 +/- 0.9% DeltaAAD) and kinetics (-0.01% DeltaAAD/s) of the TGF-independent system were markedly blunted whereas the TGF-intact system exhibited continued vasoconstriction with slower kinetics (-0.20% DeltaAAD/s) until a steady-state plateau was reached (-25.9 +/- 0.4% DeltaAAD). Thus the TGF mechanism plays a role in both direct mediation of vasoconstriction and in modulation of the myogenic response.

Coupling of M(2) muscarinic receptors to ERK MAP kinases and caldesmon phosphorylation in colonic smooth muscle.

Coupling of M(2) and M(3) muscarinic receptors to activation of mitogen-activated protein (MAP) kinases and phosphorylation of caldesmon was studied in canine colonic smooth muscle strips in which M(3) receptors were selectively inactivated by N, N-dimethyl-4-piperidinyl diphenylacetate (4-DAMP) mustard (40 nM). ACh elicited activation of extracellular signal-regulated kinase (ERK) 1, ERK2, and p38 MAP kinases in control muscles and increased phosphorylation of caldesmon (Ser(789)), a putative downstream target of MAP kinases. Alkylation of M(3) receptors with 4-DAMP had only a modest inhibitory effect on ERK activation, p38 MAP kinase activation, and caldesmon phosphorylation. Subsequent treatment with 1 microM AF-DX 116 completely prevented activation of ERK and p38 MAP kinase and prevented caldesmon phosphorylation. Caldesmon phosphorylation was blocked by the MAP kinase/ERK kinase inhibitor PD-98509 but not by the p38 MAP kinase inhibitor SB-203580. These results indicate that colonic smooth muscle M(2) receptors are coupled to ERK and p38 MAP kinases. Activation of ERK, but not p38 MAP kinases, results in phosphorylation of caldesmon in vivo, which is a novel function for M(2) receptor activation in smooth muscle.

Postglomerular vasoconstriction to angiotensin II and norepinephrine depends on intracellular calcium release.

The current study was performed to determine the effect of calcium store depletion with cyclopiazonic acid (CPA) on the pre- and postglomerular vasoconstrictor responses to angiotensin II (ANG II) and norepinephrine (NE). CPA treatment significantly attenuated the afferent arteriolar response to 10 nM ANG II by 51% and to 1000 nM NE by 19%. Efferent arteriolar responses to ANG II and NE were also greatly attenuated in the presence of CPA. These data demonstrate that afferent and efferent arteriolar responses to ANG II and NE depend on release of calcium from CPA-sensitive intracellular stores. Furthermore, the postglomerular response to these agents exhibits a greater dependency on calcium release from intracellular stores.

Assessing the pain of people with cognitive impairment.

This review presents evidence of the undertreatment of pain for people with cognitive impairment and explores reasons for this, emphasizing inadequate detection due to lack of suitable pain assessment protocols. Implications for practice and suggestions for further research are made.

Effects of glutamine supplementation of an amino acid-based purified diet on intestinal mucosal integrity in cats with methotrexate-induced enteritis.

OBJECTIVE: To determine effects of glutamine-supplemented and glutamine-free amino acid-based purified diets, compared with a dry expanded diet, on intestinal structure and function in a model that used cats with methotrexate-induced enteritis. ANIMALS: 18 adult specific-pathogen-free cats. PROCEDURE: 12 cats were given intragastric feedings of an amino acid-based purified diet supplemented with glutamine (7% [wt:wt]) or an isonitrogenous amount of glycine and alanine; 6 cats consumed a dry expanded diet. After 21 days, cats received methotrexate (MTX; 11 mg/kg of body weight, IV). Intestinal permeability testing was performed immediately before and 66 hours after MTX administration. Celiotomy was performed 72 hours after MTX administration for aseptic removal of mesenteric lymph nodes, collection of full-thickness intestinal biopsy specimens, determination of intestinal cellular proliferation, and collection of aortic and portal venous blood samples for determination of arteriovenous amino acid concentrations across the intestine. RESULTS: Administration of MTX was associated with severe enterotoxicosis manifested as diarrhea (8/12 cats), vomiting (12/12), and positive results for bacterial culture of mesenteric lymph nodes (12/12) in cats receiving the purified diets, independent of glutamine supplementation. Diet did not affect villus tip length and villus surface area in the small intestine or cellular proliferation. Administration of MTX was associated with significantly increased intestinal permeability, which was not attenuated by glutamine supplementation. CONCLUSIONS: Feeding of a glutamine-supplemented amino acid-based purified diet was unable to preserve intestinal function in cats with MTX-induced enteritis. CLINICAL RELEVANCE: Intestinal morphologic alterations correlate poorly with intestinal function as measured by means of bacterial translocation and intestinal permeability.

Candesartan cilexetil protects against loss of autoregulatory efficiency in angiotensin II-infused rats.

Renal autoregulatory efficiency is compromised in angiotensin-II (AngII)-dependent Goldblatt hypertension. The current studies were performed to assess renal autoregulatory capability in AngII-infused hypertensive rats and to determine the effect of chronic candesartan cilexetil treatment on autoregulatory behavior. Rats received chronic infusion of AngII (60 ng/min) or vehicle via an osmotic minipump implanted subcutaneously in the dorsum of the neck. Selected rats received the novel AT1 receptor blocker candesartan cilexetil (1.0 mg/kg per d) in the drinking water. Systolic BP averaged 118+/-1 mmHg (n=34) before pump implantation. Chronic AngII infusion for 6 d increased arterial pressure to 151+/-4 mmHg. Candesartan cilexetil administration prevented the AngII-dependent increase in systolic BP. Microvascular autoregulation experiments were performed in vitro using the blood-perfused juxtamedullary nephron technique combined with videomicroscopy. Renal perfusion pressure was set at 100 mmHg during the control period before being decreased to 65 mmHg. Afferent arteriolar diameter was measured continuously as the perfusion pressure was increased from 65 mmHg to 170 mmHg in 15-mmHg increments. Afferent arteriolar diameter in sham-treated rats was 120% of control at a perfusion pressure of 65 mmHg and decreased to 76% of the control diameter at 170 mmHg (n=6). This behavior is consistent with normal autoregulatory behavior. Arterioles from rats receiving chronic infusion of AngII exhibited compromised renal microvascular autoregulatory efficiency. Afferent arteriolar diameter in AngII-treated kidneys varied from 103 to 100% (n=6) of the control diameter over the same pressure range of 65 to 170 mmHg. This blunting of autoregulatory behavior was prevented by AT1 receptor blockade. In animals receiving AngII + candesartan cilexetil, stepwise changes in perfusion pressure elicited changes in afferent arteriolar diameter between 120 and 84% after 6 d of treatment (n=6). These data suggest that chronic elevations in circulating AngII and/or the associated increase in arterial pressure impairs renal autoregulatory capability. Furthermore, inhibition of AT1 receptors with candesartan cilexetil provides protection against AngII-mediated increases in arterial pressure and prevents the associated deterioration of renal autoregulatory responsiveness.

Direct assessment of renal microvascular responses to P2-purinoceptor agonists.

Studies were performed to determine the responsiveness of rat juxtamedullary afferent arterioles to receptor-selective P2-purinoceptor agonists. Experiments were performed in vitro using the blood perfused juxtamedullary nephron technique, combined with videomicroscopy. Renal perfusion pressure was set at 110 mmHg and held constant. Basal afferent arteriolar diameter averaged 22.0 +/- 0.6 microns (n = 69). Stimulation with 0.1, 1.0, 10, and 100 microM ATP (n = 10) elicited a concentration-dependent vasoconstriction averaging 8 +/- 2, 17 +/- 2, 21 +/- 4, and 23 +/- 5%, respectively. A nearly identical afferent arteriolar vasoconstriction was observed in response to the P2X-selective agonist beta,gamma-methylene ATP (n = 10); however, another P2X agonist, alpha,beta-methylene ATP, evoked marked receptor desensitization (n = 10). Vessel diameter decreased by approximately 7 +/- 2, 16 +/- 2, 23 +/- 3, and 22 +/- 3%, respectively, over the same concentration range. The P2Y-selective agonist, 2-methylthio-ATP, evoked only a modest vasoconstriction, whereas UTP and adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S) reduced afferent diameter markedly at concentrations > 1.0 microM. Afferent arteriolar diameter decreased by 5 +/- 4, 31 +/- 8, and 72 +/- 8% during UTP administration (n = 7) at concentrations of 1.0, 10, and 100 microM, respectively. Similarly, ATP gamma S (n = 6) decreased afferent diameter by 16 +/- 2, 58 +/- 8, and 98 +/- 3%, respectively, over the same concentration range. Nitric oxide synthesis inhibition with N omega-nitro-L-arginine did not significantly alter the afferent arteriolar response to ATP but did potentiate ATP-mediated arcuate artery vasoconstriction. The following data suggest the presence of multiple P2 receptors on juxtamedullary afferent arterioles and are consistent with classification of those receptors as members of the P2X- and P2Y2 (P2U)-receptor subtypes.

Calcium mobilization contributes to pressure-mediated afferent arteriolar vasoconstriction.

Preglomerular responses to vasoactive agonists utilize calcium released from intracellular stores and activation of calcium influx pathways to elicit vasoconstriction. The current study was performed to determine the role of calcium release from intracellular stores on the afferent arteriolar response to increases in perfusion pressure. Experiments were performed, in vitro, using the blood perfused, juxtamedullary nephron technique combined with videomicroscopy. The response of afferent arterioles to 30 mm Hg increases in perfusion pressure was determined before and after depletion of intracellular calcium pools with a 10-minute preincubation with 1 micromol/L thapsigargin or 100 micromol/L cyclopiazonic acid. Afferent arteriolar diameter averaged 20.2+/-1.0 microm (n=19) at a control perfusion pressure of 100 mm Hg. Increasing perfusion pressure to 130 and 160 mm Hg reduced afferent caliber by 10.7+/-1.0% (P<.05 versus con) and by 24.7+/-1.6% (P<.05 versus diameter at 130 mm Hg); respectively. Thapsigargin significantly increased afferent diameter by 21+/-2% (n=6) at 100 mm Hg and prevented pressure-induced autoregulatory responses. Afferent diameter averaged 24.3+/-1.7, 24.5+/-1.8 and 24.3+/-1.8 microm at perfusion pressures of 100, 130 and 160 mm Hg; respectively. Cyclopiazonic acid treatment also inhibited autoregulatory behavior but did not alter resting vessel diameter. Afferent arteriolar diameter (n=6) averaged 21.4+/-1.9 microm at 100 mm Hg and 20.9+/-2.1 and 20.5+/-2.2 microm at 130 and 160 mm Hg; respectively. Additional studies were performed to assess the role of phospholipase C activity in pressure-mediated autoregulatory behavior of afferent arterioles. Step increases in perfusion pressure decreased afferent diameter by 10.7+/-3.8 and 21.7+/-4.1%; respectively. Administration of the phospholipase C inhibitor, U-73122, (5 micromoles/L) did not significantly alter baseline diameter but did attenuate the pressure-mediated vasoconstrictor response. Increasing perfusion pressure to 130 and 160 mm Hg reduced afferent diameter by only 6.5+/-1.5 and 10.0+/-2.0%; respectively. These data demonstrate that interruption of calcium mobilization with thapsigargin, cyclopiazonic acid, or phospholipase C inhibition markedly attenuates pressure-mediated afferent arteriolar vasoconstriction and suggests that autoregulatory adjustments in afferent arteriolar diameter involve calcium release from inositoltrisphosphate(IP3)-sensitive intracellular stores.

Dietary modulation of methotrexate-induced enteritis in cats.

OBJECTIVE: To determine effects of purified and dry expanded (complex) diets on intestinal structure and function in healthy cats and in a feline model of methotrexate-induced enteritis. ANIMALS: 19 adult specific-pathogen-free cats. PROCEDURE: Cats were randomized in groups to receive a purified diet intragastrically or a complex diet orally to meet their daily metabolizable energy requirements. After 21 days, cats received either methotrexate (MTX; 10 mg/kg of body weight, i.v., n = 12) or saline solution i.v. (n = 7), and were anesthetized 72 hours later. Celiotomy was performed for aseptic removal of mesenteric lymph nodes, full-thickness biopsy of the gastrointestinal tract, and collection of aortic and portal venous blood samples for determination of arteriovenous amino acid concentrations across the intestine. RESULTS: MTX was associated with severe enterotoxicosis in cats receiving the purified diet, as manifested by diarrhea (4 of 6 cats) and vomiting (2 of 6 cats). One cat receiving the complex diet developed mild diarrhea, and none of these cats vomited. The purified diet was associated with marked villus blunting in the proximal and distal portions of the duodenum and increased bacterial translocation (3 of 6 cats), whereas none of the cats in the complex diet group developed bacterial translocation after MTX administration. For the cats given saline solution, bacterial translocation occurred in 1 of 4 cats receiving the complex diet versus 2 of 3 cats receiving the purified diet. CONCLUSIONS: Feeding of a complex diet containing intact protein as the nitrogen source abrogated the proximal small intestinal atrophy and bacterial translocation associated with feeding an amino acid-based purified diet. CLINICAL RELEVANCE: Use of purified diets containing free amino acids as the only nitrogen source cannot be endorsed in human and animal cancer patients receiving systemic chemotherapy.

Afferent and efferent arteriolar vasoconstriction to angiotensin II and norepinephrine involves release of Ca2+ from intracellular stores.

Renal vascular responses to angiotensin II (Ang II) and norepinephrine (NE) are reported to involve both mobilization of calcium from intracellular stores and activation of calcium influx pathways. The present study was conducted to determine the contribution of calcium release from intracellular stores to afferent and efferent arteriolar responses to Ang II and NE. Experiments were performed in vitro using the blood-perfused, juxtamedullary nephron technique combined with videomicroscopy. The responses of afferent and efferent arterioles to Ang II and NE were determined before and after depletion of intracellular calcium pools with 1 mumol/L thapsigargin. Positive control responses were obtained with 55 mmol/L KCI. Ang II concentrations of 0.1, 1.0, and 10 nmol/L decreased afferent arteriolar diameter by 10 +/- 4%, 17 +/- 4%, and 29 +/- 6%, respectively (P < .05; n = 8). NE also decreased afferent diameter by 5 +/- 1%, 13 +/- 1%, and 57 +/- 9% at concentrations of 10, 100, and 1000 nmol/L, respectively (P < .05; n = 6). Thapsigargin treatment shifted the afferent arteriolar concentration-response curves for both Ang II and NE significantly to the right. Nevertheless, KCI evoked a pronounced vasoconstriction and decreased afferent diameter by 56 +/- 7% (P < .05; n = 6). Postglomerular responses to Ang II and NE were abolished by thapsigargin. During the control period, efferent diameter decreased by 3 +/- 1%, 7 +/- 2%, and 14 +/- 4% for the three Ang II concentrations and 3 +/- 1%, 5 +/- 1%, and 15 +/- 4% in response to the three NE concentrations, respectively. These responses were completely eliminated in the presence of thapsigargin, whereas KCI evoked an efferent arteriolar vasoconstriction of 57 +/- 9% (P < .05). These data demonstrate that agonist-induced calcium release from intracellular stores represents an essential component in the afferent and efferent arteriolar response to Ang II and NE. Furthermore, they suggest that efferent arteriolar responses to these agents may rely more heavily on calcium release from this store, whereas afferent responses may include activation of other pathways.