Outcome of bilateral equid laparoscopic ovariectomies.

OBJECTIVE: To assess outcomes and behavior changes associated with bilateral laparoscopic ovariectomies. STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Fifty-one equids. METHODS: Medical records were evaluated from equid bilateral laparoscopic ovariectomies from January 2012 to October 2018 with a potential of 6 months follow-up. Follow-up information obtained by telephone interviews included behavior before and after surgery. Likelihood ratio chi-square tests and odds ratios (OR) with 95% CI were calculated where applicable, with statistical significance at p < .05. RESULTS: Bilateral ovariectomy was performed in 51 cases, with elective (no pathologic ovaries) ovariectomies performed in 41/51 cases. Occasional estrus-like behavior was observed postoperatively in 14/51 (27%) mares, but the behavior was mild and manageable in all cases. There was no age effect on outcome in all bilateral (p = .56) or elective only (p = .36) cases. In 37/41 (90%) elective cases, improvement was observed in the reason for presentation. Some response to altrenogest administration for behavior modification was observed preoperatively in 12/18 (67%) elective cases. Response to altrenogest was not associated with (p = .31) or able to predict a beneficial response to surgery (OR = 5.5; 95% CI = 0.38-78.57; p = .21). CONCLUSION: Response to altrenogest in elective cases may not predict behavioral outcome with ovariectomy. Occasional estrus-like behavior in mares postoperatively was not problematic for any owners. Bilateral ovariectomy is a viable treatment option for owners seeking to alleviate undesirable behavior in mares. CLINICAL SIGNIFICANCE: This study should aid veterinarians and horse owners in case selection for bilateral ovariectomy.

Measurement of cations, anions, and acetate in serum, urine, cerebrospinal fluid, and tissue by ion chromatography.

Accurate quantification of cations and anions remains a major diagnostic tool in understanding diseased states. The current technologies used for these analyses are either unable to quantify all ions due to sample size/volume, instrument setup/method, or are only able to measure ion concentrations from one physiological sample (liquid or solid). Herein, we adapted a common analytical chemistry technique, ion chromatography and applied it to measure the concentration of cations; sodium, potassium, calcium, and magnesium (Na+ , K+ , Ca2+ , and Mg2+ ) and anions; chloride, and acetate (Cl- , - OAc) from physiological samples. Specifically, cations and anions were measured in liquid samples: serum, urine, and cerebrospinal fluid, as well as tissue samples: liver, cortex, hypothalamus, and amygdala. Serum concentrations of Na+ , K+ , Ca2+ , Mg2+ , Cl- , and - OAc (mmol/L): 138.8 ± 4.56, 4.05 ± 0.21, 4.07 ± 0.26, 0.98 ± 0.05, 97.7 ± 3.42, and 0.23 ± 0.04, respectively. Cerebrospinal fluid concentrations of Na+ , K+ , Ca2+ , Mg2+ , Cl- , and - OAc (mmol/L): 145.1 ± 2.81, 2.41 ± 0.26, 2.18 ± 0.38, 1.04 ± 0.11, 120.2 ± 3.75, 0.21 ± 0.05, respectively. Tissue Na+ , K+ , Ca2+ , Mg2+ , Cl- , and - OAc were also measured. Validation of the ion chromatography method was established by comparing chloride concentration between ion chromatography with a known method using an ion selective chloride electrode. These results indicate that ion chromatography is a suitable method for the measurement of cations and anions, including acetate from various physiological samples.

Expression of Proinflammatory Cytokines Is Upregulated in the Hypothalamic Paraventricular Nucleus of Dahl Salt-Sensitive Hypertensive Rats.

Accumulating evidence indicates that inflammation is implicated in hypertension. However, the role of brain proinflammatory cytokines (PICs) in salt sensitive hypertension remains to be determined. Thus, the objective of this study was to test the hypothesis that high salt (HS) diet increases PICs expression in the paraventricular nucleus (PVN) and leads to PVN neuronal activation. Eight-week-old male Dahl salt sensitive (Dahl S) rats, and age and sex matched normal Sprague Dawley (SD) rats were divided into two groups and fed with either a HS (4% NaCl) or normal salt (NS, 0.4% NaCl) diet for 5 consecutive weeks. HS diet induced hypertension and significantly increased cerebrospinal fluid (CSF) sodium concentration ([Na+]) in Dahl S rats, but not in normal SD rats. In addition, HS diet intake triggered increases in mRNA levels and immunoreactivities of PVN PICs including TNF-α, IL-6, and IL-1ß, as well as Fra1, a chronic marker of neuronal activation, in Dahl S rats, but not in SD rats. Next, we investigated whether this increase in the expression of PVN PICs and Fra1 was induced by increased CSF [Na+]. Adult male SD rats were intracerebroventricular (ICV) infused with 8 µl of either hypertonic salt (4 µmol NaCl), mannitol (8 µmol, as osmolarity control), or isotonic salt (0.9% NaCl as vehicle control). Three hours following the ICV infusion, rats were euthanized and their PVN PICs expression was measured. The results showed that central administration of hypertonic saline in SD rats significantly increased the expression of PICs including TNF-α, IL-6, and IL-1ß, as well as neuronal activation marker Fra1, compared to isotonic NaCl controls and osmolarity controls. Finally, we tested whether the increase in PICs expression occurred in neurons. Incubation of hypothalamic neurons with 10 mM NaCl in a culture medium for 6 h elicited significant increases in TNF-α, IL-6, and Fra1 mRNA levels. These observations, coupled with the important role of PICs in modulating neuronal activity and stimulating vasopressin release, suggest that HS intake induces an inflammatory state in the PVN, which, may in turn, augments sympathetic nerve activity and vasopressin secretion, contributing to the development of salt sensitive hypertension.

The Orexin System and Hypertension.

In this review, we focus on the role of orexin signaling in blood pressure control and its potential link to hypertension by summarizing evidence from several experimental animal models of hypertension. Studies using the spontaneously hypertensive rat (SHR) animal model of human essential hypertension show that pharmacological blockade of orexin receptors reduces blood pressure in SHRs but not in Wistar-Kyoto rats. In addition, increased activity of the orexin system contributes to elevated blood pressure and sympathetic nerve activity (SNA) in dark-active period Schlager hypertensive (BPH/2J) mice, another genetic model of neurogenic hypertension. Similar to these two models, Sprague-Dawley rats with stress-induced hypertension display an overactive central orexin system. Furthermore, upregulation of the orexin receptor 1 increases firing of hypothalamic paraventricular nucleus neurons, augments SNA, and contributes to hypertension in the obese Zucker rat, an animal model of obesity-related hypertension. Finally, we propose a hypothesis for the implication of the orexin system in salt-sensitive hypertension. All of this evidence, coupled with the important role of elevated SNA in increasing blood pressure, strongly suggests that hyperactivity of the orexin system contributes to hypertension.

Increased activity of the orexin system in the paraventricular nucleus contributes to salt-sensitive hypertension.

The orexin system is involved in arginine vasopressin (AVP) regulation, and its overactivation has been implicated in hypertension. However, its role in salt-sensitive hypertension (SSHTN) is unknown. Here, we tested the hypothesis that hyperactivity of the orexin system in the paraventricular nucleus (PVN) contributes to SSHTN via enhancing AVP signaling. Eight-week-old male Dahl salt-sensitive (Dahl S) and age- and sex-matched Sprague-Dawley (SD) rats were placed on a high-salt (HS; 8% NaCl) or normal-salt (NS; 0.4% NaCl) diet for 4 wk. HS intake did not alter mean arterial pressure (MAP), PVN mRNA levels of orexin receptor 1 (OX1R), or OX2R but slightly increased PVN AVP mRNA expression in SD rats. HS diet induced significant increases in MAP and PVN mRNA levels of OX1R, OX2R, and AVP in Dahl S rats. Intracerebroventricular infusion of orexin A (0.2 nmol) dramatically increased AVP mRNA levels and immunoreactivity in the PVN of SD rats. Incubation of cultured hypothalamus neurons from newborn SD rats with orexin A increased AVP mRNA expression, which was attenuated by OX1R blockade. In addition, increased cerebrospinal fluid Na+ concentration through intracerebroventricular infusion of NaCl solution (4 µmol) increased PVN OX1R and AVP mRNA levels and immunoreactivity in SD rats. Furthermore, bilateral PVN microinjection of the OX1R antagonist SB-408124 resulted in a greater reduction in MAP in HS intake (-16 ± 5 mmHg) compared with NS-fed (-4 ± 4 mmHg) anesthetized Dahl S rats. These results suggest that elevated PVN OX1R activation may contribute to SSHTN by enhancing AVP signaling.NEW & NOTEWORTHY To our best knowledge, this study is the first to investigate the involvement of the orexin system in salt-sensitive hypertension. Our results suggest that the orexin system may contribute to the Dahl model of salt-sensitive hypertension by enhancing vasopressin signaling in the hypothalamic paraventricular nucleus.

High Salt Intake Augments Excitability of PVN Neurons in Rats: Role of the Endoplasmic Reticulum Ca2+ Store.

High salt (HS) intake sensitizes central autonomic circuitry leading to sympathoexcitation. However, its underlying mechanisms are not fully understood. We hypothesized that inhibition of PVN endoplasmic reticulum (ER) Ca2+ store function would augment PVN neuronal excitability and sympathetic nerve activity (SNA). We further hypothesized that a 2% (NaCl) HS diet for 5 weeks would reduce ER Ca2+ store function and increase excitability of PVN neurons with axon projections to the rostral ventrolateral medulla (PVN-RVLM) identified by retrograde label. PVN microinjection of the ER Ca2+ ATPase inhibitor thapsigargin (TG) increased SNA and mean arterial pressure (MAP) in a dose-dependent manner in rats with a normal salt (NS) diet (0.4%NaCl). In contrast, sympathoexcitatory responses to PVN TG were significantly (p < 0.05) blunted in HS treated rats compared to NS treatment. In whole cell current-clamp recordings from PVN-RVLM neurons, graded current injections evoked graded increases in spike frequency. Maximum discharge was significantly augmented (p < 0.05) by HS diet compared to NS group. Bath application of TG (0.5 µM) increased excitability of PVN-RVLM neurons in NS (p < 0.05), yet had no significant effect in HS rats. Our data indicate that HS intake augments excitability of PVN-RVLM neurons. Inhibition of the ER Ca2+-ATPase and depletion of Ca2+ store likely plays a role in increasing PVN neuronal excitability, which may underlie the mechanisms of sympathoexcitation in rats with chronic HS intake.

Activation of the (pro)renin receptor in the paraventricular nucleus increases sympathetic outflow in anesthetized rats.

Previous studies have indicated that hyperactivity of brain prorenin receptors (PRR) is implicated in neurogenic hypertension. However, the role of brain PRR in regulating arterial blood pressure (ABP) is not well understood. Here, we test the hypothesis that PRR activation in the hypothalamic paraventricular nucleus (PVN) contributes to increased sympathetic nerve activity (SNA). In anaesthetized adult Sprague-Dawley (SD) rats, bilateral PVN microinjection of human prorenin (2 pmol/side) significantly increased splanchnic SNA (SSNA; 71 ± 15%, n = 7). Preinjection of either prorenin handle region peptide, the PRR binding blocker (PRRB), or tiron (2 nmol/side), the scavenger of reactive oxygen species (ROS), significantly attenuated the increase in SSNA (PRRB: 32 ± 5% vs. control, n = 6; tiron: 8 ± 10% vs. control, n = 5; P < 0.05) evoked by prorenin injection. We further investigated the effects of PRR activation on ROS production as well as downstream gene expression using cultured hypothalamus neurons from newborn SD rats. Incubation of brain neurons with human prorenin (100 nM) dramatically enhanced ROS production and induced a time-dependent increase in mRNA levels of inducible nitric oxide synthase (iNOS), NAPDH oxidase 2 subunit cybb, and FOS-like antigen 1 (fosl1), a marker for neuronal activation and a component of transcription factor activator protein-1 (AP-1). The maximum mRNA increase in these genes occurred 6 h following incubation (iNOS: 201-fold; cybb: 2 -fold; Ffosl1: 11-fold). The increases in iNOS and cybb mRNA were not attenuated by the AT1 receptor antagonist losartan but abolished by the AP-1 blocker curcumin. Our results suggest that PVN PRR activation induces sympathoexcitation possibly through stimulation of an ANG II-independent, ROS-AP-1-iNOS signaling pathway.

Sympathoexcitation in ANG II-salt hypertension involves reduced SK channel function in the hypothalamic paraventricular nucleus.

Hypertension (HTN) resulting from subcutaneous infusion of ANG II and dietary high salt (HS) intake involves sympathoexcitation. Recently, we reported reduced small-conductance Ca(2+)-activated K(+) (SK) current and increased excitability of presympathetic neurons in the paraventricular nucleus (PVN) in ANG II-salt HTN. Here, we hypothesized that ANG II-salt HTN would be accompanied by altered PVN SK channel activity, which may contribute to sympathoexcitation in vivo. In anesthetized rats with normal salt (NS) intake, bilateral PVN microinjection of apamin (12.5 pmol/50 nl each), the SK channel blocker, remarkably elevated splanchnic sympathetic nerve activity (SSNA), renal sympathetic nerve activity (RSNA), and mean arterial pressure (MAP). In contrast, rats with ANG II-salt HTN demonstrated significantly attenuated SSNA, RSNA, and MAP (P < 0.05) responses to PVN-injected apamin compared with NS control rats. Next, we sought to examine the individual contributions of HS and subcutaneous infusion of ANG II on PVN SK channel function. SSNA, RSNA, and MAP responses to PVN-injected apamin in rats with HS alone were significantly attenuated compared with NS-fed rats. In contrast, sympathetic nerve activity responses to PVN-injected apamin in ANG II-treated rats were slightly attenuated with SSNA, demonstrating no statistical difference compared with NS-fed rats, whereas MAP responses to PVN-injected apamin were similar to NS-fed rats. Finally, Western blot analysis showed no statistical difference in SK1-SK3 expression in the PVN between NS and ANG II-salt HTN. We conclude that reduced SK channel function in the PVN is involved in the sympathoexcitation associated with ANG II-salt HTN. Dietary HS may play a dominant role in reducing SK channel function, thus contributing to sympathoexcitation in ANG II-salt HTN.

Carpal valgus in llamas and alpacas: Retrospective evaluation of patient characteristics, radiographic features and outcomes following surgical treatment.

This study evaluated outcomes of surgical treatment for carpal valgus in New World camelids and correlated successful outcome (absence of carpal valgus determined by a veterinarian) with patient characteristics and radiographic features. Univariable and multivariable analyses of retrospective case data in 19 camelids (33 limbs) treated for carpal valgus between 1987 and 2010 revealed that procedures incorporating a distal radial transphyseal bridge were more likely (P = 0.03) to result in success after a single surgical procedure. A greater degree of angulation (> 19°, P = 0.02) and younger age at surgery (< 4 months, P = 0.03) were associated with unsuccessful outcome. Overall, 74% of limbs straightened, 15% overcorrected, and 11% had persistent valgus following surgical intervention. To straighten, 22% of limbs required multiple procedures, not including implant removal. According to owners, valgus returned following implant removal in 4 limbs that had straightened after surgery.

Valgus carpien chez les lamas et les alpagas : évaluation rétrospective des caractéristiques des patients, des radiographies et des résultats après un traitement chirurgical. Cette étude a évalué les résultats du traitement chirurgical pour le valgus carpien chez des camélidés du nouveau monde et a établi une corrélation entre les résultats fructueux (absence de valgus carpien déterminée par un vétérinaire) et les caractéristiques du patient et des radiographies. Des analyses à variable unique et à variables multiples des données de cas rétrospectifs chez 19 camélidés (33 jambes) traités pour le valgus carpien entre 1987 et 2010 ont révélé que les procédures intégrant un pont radial distal transphyséal présentaient une plus grande probabilité de succès (P = 0,03) après une seule intervention chirurgicale. Une déviation supérieure (> 19°, P = 0,02) et un jeune âge à la chirurgie (< 4 mois, P = 0,03) étaient associés à des échecs. Dans l'ensemble, 74 % des jambes ont été redressés, 15 % ont été corrigées et 11 % présentaient un valgus persistant après l'intervention chirurgicale. Pour le redressement, 22 % des jambes ont nécessité des procédures multiples, ce qui n'incluait pas l'enlèvement de l'implant. Selon les propriétaires, le valgus est réapparu après l'enlèvement de l'implant dans 4 jambes qui avaient été redressées après la chirurgie.(Traduit par Isabelle Vallières).

Sympathoexcitation and pressor responses induced by ethanol in the central nucleus of amygdala involves activation of NMDA receptors in rats.

The central nervous system plays an important role in regulating sympathetic outflow and arterial pressure in response to ethanol exposure. However, the underlying neural mechanisms have not been fully understood. In the present study, we tested the hypothesis that injection of ethanol in the central nucleus of the amygdala (CeA) increases sympathetic outflow, which may require the activation of local ionotropic excitatory amino acid receptors. In anesthetized rats, CeA injection of ethanol (0, 0.17, and 1.7 µmol) increased splanchnic sympathetic nerve activity (SSNA), lumbar sympathetic nerve activity (LSNA), and mean arterial pressure (MAP) in a dose-dependent manner. A cocktail containing ethanol (1.7 µmol) and kynurenate (KYN), an ionotropic excitatory amino acid receptor blocker, showed significantly blunted sympathoexcitatory and pressor responses compared with those elicited by CeA-injected ethanol alone (P < 0.01). A cocktail containing ethanol and d-2-amino-5-phosphonovalerate, an N-methyl-d-aspartate (NMDA) receptor antagonist, elicited attenuated sympathoexcitatory and pressor responses that were significantly less than ethanol alone (P < 0.01). In addition, CeA injection of acetate (0.20 µmol, n = 7), an ethanol metabolite, consistently elicited sympathoexcitatory and pressor responses, which were effectively blocked by d-2-amino-5-phosphonovalerate (n = 9, P < 0.05). Inhibition of neuronal activity of the rostral ventrolateral medulla (RVLM) with KYN significantly (P < 0.01) attenuated sympathoexcitatory responses elicited by CeA-injected ethanol. Double labeling of immune fluorescence showed NMDA NR1 receptor expression in CeA neurons projecting to the RVLM. We conclude that ethanol and acetate increase sympathetic outflow and arterial pressure, which may involve the activation of NMDA receptors in CeA neurons projecting to the RVLM.

Laparoscopic-assisted urinary bladder marsupialization in a goat that developed recurrent urethral obstruction following perineal urethrostomy.

CASE DESCRIPTION: A 5-year-old castrated male Nigerian Dwarf goat with recurrent urethral obstruction after perineal urethrostomy. CLINICAL FINDINGS: The wether goat was referred for evaluation of a perineal urethrostomy site; perineal urethrostomy had been performed 1.5 years earlier. The site was patent, but a moderate stricture was found just cranial to the perineal urethrostomy site. The goat had obstructed urine flow at the stricture site repeatedly in the previous few months. TREATMENT AND OUTCOME: Laparoscopically assisted urinary bladder marsupialization was recommended. The procedure was performed with the patient under general anesthesia in dorsal recumbency; food had been withheld for 36 hours prior to anesthesia. Ventral midline and left caudal paramedian portals were made for the laparoscopic camera and instruments. A larger incision in the right caudal paramedian portion of the abdomen was used to create the marsupialization stoma. Laparoscopic observation of anatomic structures within the abdomen was excellent. The goat recovered well from surgery and was discharged the day following surgery. Follow-up examinations were done every few months after surgery, and dermatitis of the abdominal skin secondary to urine scald was the only postoperative complication. At 9 months of follow-up, the goat continued to pass urine through the marsupialization site comfortably. CLINICAL RELEVANCE: In the goat of this report, laparoscopic urinary bladder marsupialization was a minimally invasive procedure with minimal signs of postoperative pain. The procedure was simple to perform and should be considered as an alternative to marsupialization via laparotomy.

Complications after orthopedic surgery in alpacas and llamas: 24 cases (2000-2006).

OBJECTIVE: To report complications associated with orthopedic surgery in alpacas and llamas. STUDY DESIGN: Retrospective study. ANIMALS: Alpacas (n=18) and llamas (n=6) that orthopedic surgery using internal or external fixation. METHODS: Medical records (January 2000-December 2006) and radiographs were reviewed and owners contacted for follow-up information for alpacas and llamas that had orthopedic surgery involving internal or external fixation. Fourteen camelids had internal fixation, 7 had external fixation, and 3 had a combination of internal and external fixation. RESULTS: Twenty-two animals (92%) were discharged after surgery (mean hospitalization, 15 days). Of 20 animals with >or=1 year follow-up information, 18 were alive (82%). Postoperative complications related to fracture healing, infection, soft tissue structures, or joints occurred in 21 camelids (87%). Thirteen animals returned to their intended use, 4 animals returned to breeding but not their intended use, 4 were euthanatized, and 3 were only able to be used as pets. CONCLUSIONS: Fixation type (internal, external) did not have any significant effect on complications involving fracture healing, infection, soft tissue structures, or chronic lameness. Camelids with open fractures were more likely to have complications associated with fracture healing, repair, and infection than closed fractures. CLINICAL RELEVANCE: Complications after orthopedic surgery in alpacas and llamas are more common than previously reported and may result in chronic lameness or prevent return to their intended use.

Tibial plateau leveling osteotomy in a llama with a ruptured cranial cruciate ligament.

A 3-year-old 155-kg (342-lb) castrated male llama was examined because of left hind limb lameness of acute onset. A diagnosis of cranial cruciate ligament and medial collateral ligament rupture was made, and tibial plateau leveling osteotomy was recommended. The tibial plateau leveling osteotomy procedure was performed as described for dogs, except that 2 orthopedic plates were used to stabilize the osteotomy because of the size of the llama. The medial collateral ligament was sutured and reinforced with 2 strands of size-2 polypropylene placed in a figure-8 fashion between cancellous bone screws in the femur and tibia. Four days after surgery, failure of the medial collateral ligament repair was evident. Approximately 3.5 years after surgery, the llama was reexamined. The owners reported that the llama had full use of its left hind limb, and only mild lameness (grade 1 of 5) was evident. Results suggest that tibial plateau leveling osteotomy may be applicable in camelids with rupture of the cranial cruciate ligament. However, additional study is needed before tibial plateau leveling osteotomy can be routinely recommended. In particular, additional information is needed on the tibial plateau slope in healthy camelids, the role of the fibula in tibial plateau leveling osteotomy procedures, and the prevalence of cranial cruciate ligament rupture in camelids.