Can peri-surgical electroacupuncture relieve immunity suppression? A pilot study in dogs.

General anesthesia and surgical stress can suppress the immunological response by acting both directly on the immune system and indirectly on the hypothalamic-pituitary-adrenal axis and the sympathetic nervous system. Disturbance of the immune system during the perioperative period can lead to complications such as wound-healing disorders and infections up to sepsis. Effectiveness of acupuncture in regulating the immune function by increasing leukocyte numbers and inhibiting inflammatory response has been proven. This study aimed to explore the impact of electroacupuncture (EAP) on the dynamic balance of the immune system and immune cell populations in dogs undergoing surgery. Twelve healthy bitches scheduled for elective ovariectomy were divided into two groups according to whether (EAP, n=6) or not (CTR, n=6) a peri-operative electroacupuncture treatment was performed. Levels of leukocytes (neutrophils, monocytes, T- and B-cells) and immunoglobulins M (IgM) and A (IgA) were measured in blood samples collected before (T0), 1 h (T1) and 2.5 h (T2) after anesthesia induction. Leukocytes count decreased from T0 to T1 in both groups and restored within 1.5 h in EAP group whereas remained significantly lower in CTR group (P<0.02). In particular, neutrophils and monocytes increased in dogs receiving EAP (P<0.01) while T-cells decreased in CTR group (P<0.04) at T2. B-cells and cytotoxic T-cells decreased in EAP dogs (P<0.04) at T2. No differences in helper T-cells, IgM and IgA levels were recorded between groups and over time. Our results suggest a modulatory effect of EAP on the immune system which is early expressed on neutrophils, monocytes and T-cells.

Solitary and multiple simultaneous malignant epithelial mammary tumours in dogs: An explorative retrospective study.

Canine mammary tumours represent a hard-prognostic task for veterinary clinicians. TNM staging and grading systems refer to a single tumour. Significant limits come to light when these systems are applied to multiple mammary tumours due to the arbitrary criterion in determining which single tumour is representative of the patient's prognosis. This study explored some clinical features of 50 dogs affected by at least one malignant mammary tumour. Clinical features and staging, together with histological classification and grading, have been related to disease-free survival (DFS) with the purpose to evaluate their impact on prognosis. The prognosis was worse in 10-11-year-old dogs (P < 0.05), in dogs affected by complex carcinoma (P < 0.05), and in patients assigned to Peña grade I (P < 0.05). The bodyweight was not linearly related to DFS (P < 0.01), and patients with a low number of neoformations (n ≤ 2) showed a better prognosis than dogs with 3-5 tumours (P < 0.05). Both the average and the total size of malignant tumours were related to DFS (P < 0.05). Dogs assigned with stage I had the best DFS (P < 0.05). In conclusion, the Peña grade I alone would not seem to guarantee a favourable prognosis when applied to mammary tumours in dogs affected by multiple simultaneous presentations. Different characteristics, besides tumour grading, such as tumour immunophenotype and expression of hormonal receptors, could in the future, contribute to elucidate the clinical behaviour of multiple canine mammary tumours.

Ovarian Embryonal Carcinoma in a Dog.

A 17-month-old female doberman pinscher was referred for an abdominal mass and ascites. Exploratory laparotomy revealed the presence of a large neoplastic mass replacing the right ovary and associated with multiple mesovarian, mesometrial and peritoneal nodules. An ovariohysterectomy was performed. Grossly, the tumour was soft and multilocular with large areas of haemorrhage and necrosis. Microscopically, it was infiltrative and composed of round and polygonal cells arranged respectively in solid sheets or forming distorted tubular structures separated by thick fibrovascular septae. Tubules contained necrotic debris, proteinaceous fluid or small endoluminal papillary structures. Marked cellular atypia, multiple neoplastic emboli and high mitotic count were observed. Immunohistochemically, the round cells uniformly expressed placental alkaline phosphatase, while the polygonal cells arranged in tubules and papillae expressed cytokeratin (CK) AE1/AE3 and CK7. A final diagnosis of metastasizing ovarian embryonal carcinoma (EC), a primitive germ cell tumour characterized by rudimentary epithelial differentiation was made. Canine ovarian EC should be considered as a differential diagnosis for undifferentiated aggressive ovarian tumours in young dogs.

Morphology and Aquaporin Immunohistochemistry of the Uterine Tube of Saanen Goats (Capra hircus): Comparison Throughout the Reproductive Cycle.

The expression of six different aquaporins (AQP1, 2, 3, 4, 5 and 9), integral membrane water channels that facilitate bi-directional passive movement of water, was investigated by immunohistochemistry in the uterine tube of pre-pubertal and adult Saanen goats (Capra hircus), comparing the different phases of the oestrous cycle. Regional morphology and secretory processes were markedly different during the goat oestrous cycle. The tested AQP molecules showed different expression patterns in comparison with already studied species. AQP1-immunoreactivity was evidenced at the endothelium of blood vessels and in nerve fibres, regardless of the tubal tract and cycle period. AQP4-immunoreactivity was shown on the lateral plasmalemma in the basal third of the epithelial cells at infundibulum and ampulla level in the cycling goats, more evidently during follicular than during luteal phase. No AQP4-immunoreactivity was noticed at the level of the isthmus region, regardless of the cycle phase. AQP5-immunoreactivity, localized at the apical surface of epithelial cells, increased from pre-puberty to adulthood. Thereafter, AQP5-immunoreactivity was prominent during the follicular phase, when it strongly decorated the apical plasmalemma of all epithelial cells at ampullary level. During luteal phase, immunoreactivity was discontinuous, being weak to strong at the apex of the secretory cells protruding into the lumen. In the isthmus region, the strongest AQP5-immunoreactivity was seen during follicular phase, with a clear localization in the apical plasmalemma of all the epithelial cells and also on the lateral plasmalemma. AQP2, 3 and 9 were undetectable all along the goat uterine tube. Likely, a collaboration of different AQP molecules sustains the fluid production in the goat uterine tube. AQP1-mediated transudation from the blood capillaries, together with permeation of the epithelium by AQP4 in the basal rim of the epithelial cells and final intervening of apical AQP5, could be involved in fluid production as well as in secretory processes.

The role of birth weight on litter size and mortality within 24h of life in purebred dogs: What aspects are involved?

In humans, scientific evidence emphasizes the role of birth weight on neonatal welfare, morbidity and mortality. In canine species, defining normal ranges of birth weight is a harder issue due to a great morphological variability in size, body weight and breed. The aim of this study was to correlate birth weight with litter size and mortality within 24h of life in 789 pups from 140 litters of purebred dogs and to investigate the aspects that might affect these factors. Birth weight was influenced by maternal size, weight and age (P<0.001). The lightest pups were from toy sized or weighing up to 10 kg bitches. Conversely, bitches aged 2-8 years whelped heavier pups than younger and older mothers. Birth weight was also related both to litter size, with heavier pups in smaller rather than in larger litters from medium sized bitches, and breed (P<0.05). Unexpectedly, birth weight did not differ between live born and stillborn pups. However, birth weight was lower in pups dying within 24h of life (P<0.05). High mortality of pups was related both to short pregnancies (P<0.05), also showing lighter litters (P<0.001), and to dystocic parturitions (P<0.001). Litter size was associated with parity, type and number of mating, and length of pregnancy (P<0.001). Low birth weight appears to predispose to early neonatal mortality suggesting a predominant role of the breed rather than size and weight in determining birth weight in pups.

Prognostic potential of amniotic fluid analysis at birth on canine neonatal outcomes.

Glucose, lactate and cortisol concentrations in amniotic fluid were measured at birth in 95 pups and related to neonatal viability based on Apgar scoring and to neonatal mortality. Neither amniotic parameters nor neonatal mortality were associated with the Apgar score. Stillborn pups showed high lactate (P < 0.001) and cortisol (P < 0.05) but low glucose amniotic concentrations (P < 0.001). No amniotic fluid differences were observed between normal and malformed pups. Amniotic glucose (P < 0.001), lactate (P < 0.05) and cortisol (P < 0.05) concentrations were higher in pups delivered by vaginal parturition than by Caesarean section. Birth weight was higher in live pups than in pups dying within 48 h (P < 0.05). Although these are preliminary results, the analysis of amniotic fluid collected at birth could be a valuable predictor of neonatal outcomes in dogs.

Periovulatory time in the bitch: what's new to know?: Comparison between ovarian histology and clinical features.

The ability to recognize specific events happening in the ovaries during periovulatory time allows optimal management of canine reproduction. The objective of this study was to assess the efficacy of vaginal cytology and blood progesterone (P4) assay to identify accurately the changes occurring at the ovarian structures, mainly during the fertile period. Tertiary follicles, corpora hemorrhagica (CHs) and corpora lutea (CLs) from forty healthy bitches undergoing ovariohysterectomy were evaluated by histo-morphometry based on their aspect, number and size. The tertiary follicles distribution (small, medium and large) was statistically different (P<0.002) among all the stages of the reproductive cycle, except for small follicles (<2mm), which were always observed from proestrus to anestrus. Very large follicles (>4mm) were predominant (P=0.008) around ovulation when P4 mean level was 6.1±1.7ng/mL. The early postovulatory estrous period was characterized by CHs (P<0.002) and P4 level of 16.7±5.9ng/mL. The end of the fertile period - start of diestrus - coincided with the development of CLs (P=0.001) associated with a P4 mean level of 73.9±9.9ng/mL. The small (P<0.001) and medium (P<0.05) follicle diameters were positively correlated with the bitch size. The number of follicles larger than 4mm was significantly lower in bitches younger than 4 years (P<0.02). This study provides insight into some critical steps in the canine reproductive processes in the periovulatory phase and the end of the fertile period, essential to plan breeding programs.

Breed-specific fetal biometry and factors affecting the prediction of whelping date in the German shepherd dog.

To date many studies have been published about predicting parturition by ultrasonographic fetal measurements in the bitch. Given that accuracy in such prediction is a key point for clinicians and breeders, formulas to calculate the whelping date were mainly obtained from small and medium sized dogs, which means poor accuracy when applied to large or giant breeds. Based on the evidence that ethnicity significantly affects fetal biometry in humans, this study aimed at developing a breed-specific linear regression model for estimating parturition date in the German shepherd dog. For this purpose, serial ultrasonographic measurements of the inner chorionic cavity diameter (ICC) and the fetal biparietal diameter (BP) were collected in 40 pregnant German shepherd bitches. The quality of the regression models for estimating parturition date was further verified in 22 other pregnant German shepherd bitches. Accuracy related to the prediction of parturition date was higher than previously reported: 94.5% and 91.7% within ±2 days interval based on ICC and BP measurements, respectively. Additional investigation was performed on the effects of maternal weight, age and litter size in relation to fetal biometry and to accuracy of parturition estimation. Moreover, the study included a comparison between hormonal and fetal ultrasound (ICC and BP) measurements connected to the estimation of whelping date. We suggest that specific equations from a single breed are likely to offer excellent accuracy, comparable to that of periovulatory progesteronemia, in parturition prediction and to avoid morphological variables present in dogs of different breeds even with the same size/weight.

Doppler ultrasonographic evaluation of ductus venosus blood flow in 55 canine fetuses.

PURPOSE: The ductus venosus (DV) blood flow has been studied in fetal lambs and in humans. This study aims to describe the velocities, the Doppler indices and the morphological patterns of the venous blood flow in the DV of canine fetuses during physiological pregnancy. METHODS: The DV of 55 canine fetuses has been evaluated and the waveforms described using B-mode, color and pulsed-wave Doppler ultrasound technology. RESULTS: We found 48 diphasic waves and 7 threephasic waves. No monophasic waveform was found. Six of seven threephasic waveforms belonged to litters in which perinatal mortality occurred. The peak velocity during ventricular systole S (cm/s), the peak velocity during the ventricular diastole D (cm/s), the velocity during atrial contraction aV (cm/s), the S/D index, the pulsatility index (PI) and the resistance index were measured. CONCLUSIONS: All Doppler indices and velocities were significantly correlated with each other (p < 0.05). The number of newborn puppies and the age of bitches were not related to DV vascular indices or flow rate (p > 0.05). Gestational age was proportional to the PI (p < 0.02). Doppler ultrasonography allows the assessment of DV blood flow in canine fetuses during pregnancy.

Expression of aquaporin water channels in canine fetal adnexa in respect to the regulation of amniotic fluid production and absorption.

Amniotic fluid (AF) is created by the flow of fluid from the fetal lung and bladder and reabsorbed in part by fetal swallowing and partly by the transfer across the amnion to the fetal circulation. Placental water flux is an important factor in determining AF volume and fetal hydration. In addition the fetal membranes might be involved in the regulation of fluid composition. To understand the mechanisms responsible for maintaining a correct balance of AF volume we evaluated the expression of aquaporins (AQPs) in canine fetal adnexa. AQPs are a family of integral membrane proteins permitting passive but physiologically rapid transcellular water movement. The presence of AQP1, 3, 5, 8 and -9 was immunohistochemically assessed in canine fetal adnexa, collected in early, middle and late-gestation during ovario-hysterectomies performed with fully informed owners' consent. Changes in AF volume and biochemical composition were also evaluated throughout pregnancy. Our results show distinct aquaporin expression patterns in maternal and extraembryonic tissues in relation to pregnancy period. AQP1 was localized in placental endothelia, allantochorion, amnion, allantois and yolk sac. AQP3 was present in the placental labyrinth, amnion, allantois and yolk sac. AQP8 was especially evident on the epithelia lining the glandular chambers, the amniotic and allantois sacs. AQP9, a channel highly permeable to water and urea, was observed in epithelia of amnion, allantois and yolk sac. In summary, AQP1, 3, 5, 8 and -9 have distinct expression patterns in canine fetal membranes and placenta in relation to pregnancy period, suggesting an involvement in mediating the AF changes during gestation.

Vaginal bacterial flora and cytology in proestrous bitches: role on fertility.

The study of canine vaginal cytology underwent limited evolution over the years. Presence and significance of inflammatory cells in vaginal smears are little considered aspects in the bitch. Moreover, occurrence of vaginal bacteria in breeding bitches during follicular phase of the reproductive cycle, in absence of clinical signs of infection, involves the difficult question of antibiotics administration. The aim of this study was to relate findings in vaginal cytology (presence of neutrophils, lymphocytes, eosinophils, erytrocytes and bacteria) and microbial environment during proestrus with fertility outcomes (development of pregnancy, uterine infection, resorption, abortion and neonatal mortality). Bacteria sensitivity to antibiotics normally used in small animal practice was also evaluated. Bacteria isolated from vagina, in order of frequency, were Enterococcus faecalis, Streptococcus ß-haemolyticus, Pasteurella multocida, E. coli, Klebsiella pneumoniae, Proteus mirabilis, E. coli haemolyticus, Arcanobacterium pyogenes, Streptococcus spp., Staphylococcus spp. and Acinetobacter spp. No mycoplasmas were observed. The present study showed that proestrous cytological aspects do not affect fertility. Eosinophils were never detected, while erythrocytes were always detected. During diestrus, E. coli was found in all pregnant bitches that developed clinical symptoms of uterine disorders (n = 3), resulting in uterine infection, resorption or abortion, but without statistical significance. Vaginal presence of Streptococcus spp. in proestrus was instead negatively associated with development of uterine infections (P = 0.005). Therefore, Streptococcus spp. could have a protective competitive role against more dangerous pathogens affecting fertility of the bitch. Among the 12 antibiotics tested, Gram-negative bacteria showed a significant sensitivity towards the amoxicillin and clavulanic acid association (P = 0.038). However, antibiotic treatment before mating, on the basis of positive culture, yet in the absence of clinical signs, seems to be unnecessary besides harmful leading to imbalance in vaginal commensal flora with adverse effects on fertility. In conclusion, vaginal bacteria, neutrophils, lymphocytes and erytrocytes should be considered as physiological aspect in the bitch during proestrus that does not require antibiotic therapy when asymptomatic.

XX SRY-negative true hermaphrodism in two dogs: clinical, morphological, genetic and cytogenetic studies.

This work aimed at giving a deeper insight into peculiar cases of intersexuality occurring in dogs and known as XX true hermaphrodism due to the existence of both testicular and ovarian tissue in one or both gonads in the presence of an XX chromosome constitution. Clinical, histological and genetic approaches were used in the study of an 8-month-old Cocker Spaniel dog and a 3-year-old mixed-breed Pitbull, both showing a female phenotype, clitoromegaly and male behavior. A normal female karyotype (2n = 78,XX) was noticed, and polymerase chain reaction failed to detect SRY in genomic DNA obtained from peripheral blood lymphocytes of both dogs. The reproductive tract was removed by standard ovariohysterectomy and processed for histology. Thereafter, a normal female phenotype was reconstructed by vaginoplasty. Histological examination revealed bilateral ovotestis in both cases: the gonads showed immature testicular parenchyma containing seminiferous tubules, Sertoli and Leydig cells, but no signs of spermatogenesis, together with differently developed ovarian follicles containing oocytes. In the ovotestes, steroidogenesis was detected by P450c17-immunoreactivity in Leydig cells as well as in theca cells, whereas no MIS-immunoreactivity was shown by the Sertoli cells. Genital tracts of Wolffian and Müllerian origin co-existed in both subjects. Both dogs belong to the very rare cases in which testicular tissue develops in the absence of the key gene, SRY. Up to date very few genetic events have been associated with this abnormal sexual differentiation: SOX9 over-expression and RSPO1 mutation. Nevertheless, neither of them has been found in these dogs.

Effectiveness of electroacupuncture analgesia compared with opioid administration in a dog model: a pilot study.

BACKGROUND: Although opioid analgesics are the usual drugs to treat post-surgical pain, acupuncture has also been demonstrated to relieve various pain syndromes. The present pilot study aims to investigate the efficacy of electroacupuncture compared with a conventional opioid compound, butorphanol, for postoperative pain treatment in dogs undergoing elective ovariohysterectomy. METHODS: Twelve dogs were randomly allocated into two groups. Dogs received either electroacupuncture stimulation (16 and 43 Hz) at Shen Shu, Chang Shu, He Gu, Tai Yuan, Zu San Li, Yang Ling Quan, and Bai Hui acupoints, while control dogs were treated with butorphanol. Cardiovascular and respiratory parameters were recorded for both groups during operation. Plasma ß-endorphin concentrations were evaluated before surgery (baseline) and up to 24 h later. For each dog, pain was measured according to a dedicated subjective pain scoring system. RESULTS: Plasma ß-endorphin levels in dogs receiving electroacupuncture increased significantly against baseline values after 1 and 3 h after surgery. Moreover, the end-tidal isoflurane concentration needed for second ovary traction was significantly lower in acupuncture-treated dogs than control animals. All animals having electroacupuncture experienced prolonged analgesia, over 24 h at least, while four out of six dogs treated with butorphanol needed post-surgical ketorolac and tramadol supplementation to their pain relief. CONCLUSIONS: The results obtained from the present investigation showed some evidence for electroacupuncture as an alternative technique to provide postoperative analgesia in dogs.

Evaluation of newborn canine viability by means of umbilical vein lactate measurement, apgar score and uterine tocodynamometry.

Newborn viability evaluation and early detection of fetal distress could contribute to reducing mortality at birth in canine species. High neonatal mortality rate in dogs is reported subsequent to complicated or uncomplicated whelping. Umbilical vein lactate and tocodynamometry could provide valuable clinical information to the obstetricians so that appropriate medical and surgical treatments or oxygen and warm administration can be properly and timely applied to mother and newborn pup. In humans, the fetal lactate level represents an objective indicator of fetal distress and a valid predictor of babies' survival. Fetal acidosis recognition by umbilical lactate (UL) measurement, APGAR score classification, and uterine activity monitoring during labour, can represent an advanced system in the evaluation of the canine newborn patient. The purpose of this study was to correlate UL levels with canine neonatal morbidity and mortality within 48 h of birth. We evaluated the relationship among neonatal parameters at birth (mucous membrane color, heart and respiratory rate, reflex irritability, mobility, suckling and vocalization, UL, weight, and temperature) with labour characteristics (uterine contractions recorded by the tocodynamometric system of Whelpwise ™ Veterinary Perinatal Specialties ®, delivery time, and pup presentation), in view to predict pup viability. We considered also vaginal parturition versus elective and emergency Caesarean section, and uterotonic drugs influence on delivery. Umbilical lactate concentration proved to be useful to predict canine neonatal mortality within 48 h of birth (P < 0.05). We identified 5 mmol/L of vein umbilical lactate concentration as the cut off value, allowing us to distinguish between healthy and distressed pups. Higher values of UL were related with distressed pups, whereas lower values characterized vigorous pups. Lactate concentrations lower than 5 mmol/L and APGAR scores higher than 9, related to mean delivery time of 105 min with effective uterine contractions (10 mm of Hg of strength or more, frequency from 4 to 12 contractions per hour, and 2-5 min in duration), should be considered good prognostic factors in canine labour and neonatology.

An insight into testis and gubernaculum dynamics of INSL3-RXFP2 signalling during testicular descent in the dog.

Insulin-like 3 (INSL3) plays a prominent role in male development and is supposed to induce the growth of the gubernaculum testis (g.t.), thus being directly involved in testicular descent in humans and rodents. This happens through activation of the RXFP2 receptor (GREAT or LGR8). The INSL3-RXFP2 complex is reputed to play an additional paracrine role in the testis, possibly acting as part of an autocrine feedback loop. The present work provides evidence of the immunolocalisation of INSL3 in the Leydig cells of canine fetuses and of the expression of RXFP2 receptor in different tissues of the g.t. of the same specimens. RXFP2 was localised at the cell membrane of g.t. muscle and connective cells, as well as in the epithelial cells of the developing excurrent ducts. Notably, RXFP2 immunoreactivity of the g.t. was limited to fetuses at ~35-45 days of gestation, which is also the fetal period when the endocrine compartment of the dog testis is active endocrinologically, as confirmed by the anti-P450c17 and anti-INSL3 immunoreactivities of the fetal Leydig cells, and by anti-Müllerian hormone immunoreactivity of the Sertoli cells. The same immunoreactivities were also evaluated in the testes of cryptorchid dogs of different ages. RXFP2 immunoreactivity was absent from genital tracts of cryptorchid testes and g.t. remnants.

Endometrial cytology and computerized morphometric analysis of epithelial nuclei: a useful tool for reproductive diagnosis in the bitch.

New diagnostic approaches are required to recognize early canine hypofertility or infertility. We suggest that the identification of different cytologic types, cellular aspects, and nuclear features of the endometrial epithelial cells may be suitable for this purpose. This study was performed on the bitch (Canis familiaris) during the physiologic reproductive cycle and in uterine diseases. We also applied computerized cytomorphometry to evaluate nuclear area, perimeter, diameter, density, aspect, and roundness of endometrial epithelial cells in healthy dogs (N=35) at different stages of the reproductive cycle (before puberty, during proestrus, estrus, diestrus, and anestrus) and in bitches affected by uterine disorders (N=10). The stage of the estrous cycle was determined by vaginal cytology and progesterone evaluation and also confirmed by clinical and histologic observations. Samples for endometrial cytology were collected in vivo by uterine flushing with transcervical uterine cannulation. After uterine sampling, each dog underwent OHE or uterine stump revision. Cytologic analyses were compared with histologic examinations to verify the uterine condition. The uterine cellular population was represented by endometrial epithelial cells, erythrocytes, neutrophils, lymphocytes, eosinophils, macrophages, plasma cells, and cervical or incidental vaginal cells. Bacteria and amorphous material were observed. The proportion of different cells and nuclear features in the cytologic samples varied throughout the stages of the reproductive cycle and between normal and pathologic uterine conditions. The computer-assisted nuclear morphometry, performed in cytologic specimens by means of the six nuclear parameters chosen to evaluate the endometrial epithelial cell population, proved to be useful for determining the stage of the reproductive cycle. Furthermore, this system was demonstrated to be a valid support to diagnose and distinguish uterine disorders.

A collaboration of aquaporins handles water transport in relation to the estrous cycle in the bitch uterus.

Fluid movement through uterine cell membranes is crucial, as it can modulate the tissue imbibition pattern in the different phases of the estrous cycle. To gain insight into the mechanisms underlying steroid-controlled water handling, the presence and distribution of aquaporins (AQPs), integral membrane channel proteins permitting rapid passive water movement, was explored in bitch uterine tissues. Immunohistochemistry and Western immunoblot analysis were used to study the presence of AQP1, AQP2, and AQP5 in the layers of the bitch uterine wall during the different estrous phases. Presence of endothelial nitric oxide-generating enzyme NO synthase (NOS3) was also investigated, as it is known that the vasodilator NOS3 might be involved in the development of uterine edema. The results demonstrated the following: (1) AQP1, AQP2, and AQP5 were present in the uterus of cycling bitches. (2) AQP1 was localized within uterine mesometrial, myometrial, and endometrial blood vessels and in the circular and longitudinal layers of myometrium. AQP1 localization and expression were unaffected by the estrous cycle. (3) The estrogenic milieu was probably at the basis of AQP2 expression in the glandular and luminal epithelium of the endometrium. (4) AQP5 water channels were present in the apical plasma membrane of uterine epithelial cells in coincidence with plasma progesterone increase. (5) NOS3 was localized in the myometrial and epithelial tissues as well as in blood vessels indicating a contribution of this vasoactive peptide to the uterine imbibition processes. Thus, we can hypothesize that a functional and distinctive collaboration exists among diverse AQPs in water handling during the different functional uterine phases.

Mutations in the RSPO1 coding region are not the main cause of canine SRY-negative XX sex reversal in several breeds.

This report details a case of SRY-negative XX sex reversal in a mixed breed dog and surveys affected dogs of several breeds for mutations in RSPO1 coding regions. Genomic DNA from the mixed breed case was evaluated for mutations in candidate genes. Sequencing identified a homozygous G to A transition in RSPO1 exon 4 that changes a highly conserved amino acid codon in the thrombospondin domain. The possibility that this was a single nucleotide polymorphism (SNP) could not be excluded by genotyping family members. Therefore, the coding region of RSPO1 was sequenced in a survey of affected dogs, which identified a T to C transition (exon 3) in some, the above G to A transition (exon 4) in others, and no change in the remaining affected dogs. Genotypes at these base pair positions were not uniquely associated with the affected phenotype in any breed, indicating the identified transitions are most likely SNPs, not causative mutations for this canine disorder. However, the possibility that polymorphisms play a modifier role, such as changing threshold or severity of phenotypic expression in a mixed breed dog, cannot be excluded. This study emphasizes the importance of canine pedigree, breed, and population studies in evaluating candidate mutations.

Characterization of a population of unique granular lymphocytes in a bitch deciduoma, using a panel of histo- and immunohistochemical markers.

The ovaries and uterus were collected after ovariohysterectomy from a 16-month-old Labrador bitch in diestrus that never mated. Discrete swellings were found in the uterine horns, with the macroscopic appearance of normal early pregnancy. At histologic examination, the endometrium, devoid of any conceptus and chorion, showed a marked proliferation, on the basis of which a diagnosis of deciduoma was made. A remarkable population of stromal eosinophilic granular lymphocytes was present, especially in the axis of the endometrial folds. Periodic acid-Schiff and Dolichos biflorus-lectin histochemical reaction and a panel of 10 immunohistochemical markers were used to characterize eosinophilic granular cells. Our findings allowed us to compare these granular cells with the granulated decidual cells, whose presence was until now described only in primates, rodents, or a few other epitheliochorial species. On the basis of our results, the importance of eosinophilic granular cells in a decidualization process is hypothesized to occur also in the bitch.