Effect of Bladder Injection of OnabotulinumtoxinA on the Central Expression of Genes Associated with the Control of the Lower Urinary Tract: A Study in Normal Rats.

To investigate a possible central mechanism of action of Botulinum toxin A (BoNT/A) following injection in the bladder, complementary to the acknowledged peripheral bladder effect, we studied changes in the expression of neuropeptides and receptors involved in lower urinary tract function in the spinal cord (SC) and dorsal root ganglia (DRG) of normal rats following BoNT/A bladder injection. Thirty-six Sprague-Dawley rats, divided into three groups of n = 12, received bladder injections of 2U or 5U OnabotulinumtoxinA (BOTOX®), or saline. Six animals from each group were sacrificed on days 7 and 14. Expression of Tachykinin 1 (Tac1), capsaicin receptor (TRPV1), neuropeptide Y (NPY), proenkephalin (PENK) and muscarinic receptors M1, M2, M3, was evaluated in the bladder, L6-S1 DRG, and SC segments using real-time PCR and Western blotting. Real-time PCR revealed increased expression of NPY in all tissues except for SC, and increased TRPV1 and PENK expression in DRG and SC, whereas expression of Tac1, M1 and M2 was decreased. Less significant changes were noted in protein levels. These findings suggest that bladder injections of OnabotulinumtoxinA may be followed by changes in the expression of sensory, sympathetic and cholinergic bladder function regulators at the DRG/SC level.

Effect of anaesthesia on cell-mediated immunity in dogs undergoing mastectomy for mammary cancer.

OBJECTIVE: To investigate if anaesthesia for canine cancer mastectomy further influences host cell-mediated immunity (CMI) promoting cancer progression. STUDY DESIGN: A randomized, controlled, blinded clinical study. ANIMALS: A total of 20 bitches with malignant mammary tumours of clinical stage II or III undergoing the same type of mastectomy (regional mastectomy). METHODS: Dogs were randomly allocated to one of two anaesthetic groups (10 per group). The anaesthetic protocol of group A used minimally immunosuppressive drugs (tramadol, robenacoxib, propofol), whereas that of group B (control) used more immunosuppressive drugs (morphine, fentanyl, thiopental, isoflurane). For each animal, measurements of white blood cells (WBCs), neutrophils and lymphocytes, and flow cytometric assessment of T cells (CD3+), helper T cells (CD4+), cytotoxic T cells (CD8+) and CD5low+ T cells were performed prior to anaesthesia (day 0) and on days 3 and 10 postsurgery. Data were analysed using a General Linear Model for repeated measures and presented as mean ± standard deviation, p ≤ 0.05. RESULTS: In all animals, on day 3, WBCs and neutrophils were significantly increased (p < 0.0005), while flow cytometry revealed significantly decreased relative percentages of T cells (CD3+) (p = 0.003) and their subpopulations CD4+ (p = 0.006), CD8+ (p = 0.029) and CD5low+ (p = 0.031). Specifically, on day 3, the cytotoxic T cells (CD8+) were significantly decreased (p = 0.05) only in group B, whereas the CD4+ (p = 0.006) and CD5low+ (p = 0.008) T cells in group A. The only significant difference between groups was found preoperatively in the CD4+/CD8+ ratio, which was higher in group A (p = 0.006). CONCLUSIONS AND CLINICAL RELEVANCE: In dogs with mammary cancer undergoing regional mastectomy, a significant decrease in components of CMI was observed on day 3 postsurgery in both anaesthetic groups. Some indication, however, for better preserved cellular immunity by less immunosuppressive anaesthetic/analgesic drugs was detected, rendering their use advisable.

Development of real-time PCR-based methods for the detection of enzootic nasal tumor virus 2 in goats.

Enzootic nasal adenocarcinoma (ENA) is a contagious neoplasm of sheep and goats, associated with the oncogenic retroviruses enzootic nasal tumor virus (ENTV) 1 and 2, respectively. It appears to be common in countries with substantial small ruminant-production. ENA diagnosis in goats is based on autopsy and histopathology, and there is no real-time PCR method available for ENTV-2 detection. Here, a novel one-tube real-time RT-PCR (RT-qPCR) method for the detection and quantification of ENTV-2 in nasal swabs is presented. The method targets the env gene/U3 region. For the design of ENTV-2-specific oligonucleotides, molecular characterization of seven Greek ENTV-2 strains was performed. Phylogenetic analysis revealed three distinct phylogenetic clades of ENTV-2 that correlate with the country of sample collection. Evaluation of the analytical performance of the RT-qPCR revealed an amplification efficiency of 92.8% and a linear range of quantification between 2 × 108 and 2 × 102 RNA transcripts. Analysis of nasal swabs from 23 histopathologically confirmed, naturally occurring ENA cases via RT-qPCR yielded positive results. Moreover, modification of the method for use in a real-time PCR (qPCR) assay enables detection of proviral DNA in tumor specimens. Both methods are highly specific and can be used for the confirmation of ENA-suspected cases. Future applications could include ante-mortem diagnosis, verification of the ENTV-2-free status in animal trade, disease surveillance, and control programs.

Adipose-derived stromal vascular fraction aids epithelialisation and angiogenesis in an animal model.

OBJECTIVE: Limited data exist regarding the correlation between adipose-derived stromal vascular fraction (SVF) and wound healing. The aim of this study was to investigate the direct effect of intradermally injected SVF on full-thickness cutaneous wounds in a murine model. METHOD: Wistar rats were divided into three groups (A, B and C) according to their day of euthanasia (day 7, 16 and 21). Inguinal fat pad was excised and SVF enzymatically extracted. Full-thickness cutaneous wounds were created on each side of the dorsum; SVF injected intradermally at one side while the contralateral wound served as control receiving normal saline. Postoperatively, evaluation of wound healing was performed by planimetry (percentages of wound contraction, epithelialisation and total wound healing) on days 0, 3, 5, 7, 10, 13, 16 and 21, and histology and immunochemistry (cellular infiltration score, collagen production score, neoangiogenesis and epithelial thickness) on days 7, 16 and 21. RESULTS: Despite the high rate of wound contraction, it was significantly lower in the SVF-treated wounds on day 21 (p=0.037). On days 13, 16 and 21, the percentages of epithelialisation were higher in the SVF-treated wounds compared with control wounds (p=0.026, p=0.048 and p=0.05, respectively). Histologically, the number of new vessels was significantly higher in the SVF-treated wounds compared with controls on days seven (p=0.028) and 16 (p=0.027). This was also confirmed by immunohistochemistry. No significant differences were found between treated and control wounds regarding cellular infiltration score, collagen production score and epithelial thickness. CONCLUSION: Data indicate that intradermally injected SVF increases angiogenesis and enhances epithelialisation in full-thickness cutaneous wounds in rats.

Pyloric obstruction due to massive eosinophilic infiltration in a young adult dog.

A 16-month-old dog was presented with chronic vomiting, anorexia, progressive weight loss, and melena. Exploratory laparotomy revealed a massive pyloric eosinophilic infiltration leading to pyloric obstruction that was treated successfully with pylorectomy. This is a novel clinical presentation of eosinophilic gastritis and highlights the need to consider it as a differential diagnosis for pyloric obstruction.

Obstruction pylorique causée par une infiltration éosinophilique chez un jeune chien adulte. Un chien âgé de 16 mois a été présenté avec des vomissements chroniques, de l'anorexie, une perte de poids progressive et la méléna. Une laparatomie exploratoire a révélé une infiltration éosinophilique pylorique massive qui causait une obstruction pylorique qui a été traitée avec succès par la pylorectomie. C'est une présentation clinique nouvelle de la gastrite éosinophilique et elle souligne le besoin de la considérer comme un diagnostic différentiel pour l'obstruction pylorique.(Traduit par Isabelle Vallières).

First evidence of the European wildcat (Felis silvestris silvestris) as definitive host of Angiostrongylus chabaudi.

Angiostrongylus chabaudi (Strongylida, Angiostrongylidae) is a parasitic nematode described for the first time last century from the pulmonary arteries of six European wildcats (Felis silvestris silvestris) in central Italy. Since then, this parasite remained practically unknown until recently, when immature A. chabaudi have been reported from one wildcat in Germany and two domestic cats (Felis silvestris catus) in Italy. The present report describes the first record of A. chabaudi in Greece and, most importantly, the first known case of patent infection by A. chabaudi. The necropsy of a road-killed F. s. silvestris found near the lake Kerkini, in the municipality of Serres (Macedonia, Greece), revealed the presence of nematodes of both sexes in the right ventricle and the pulmonary artery of the heart. All parasites were mature adults and numerous eggs were present in the uteruses of females. The morphological characteristics of the parasites were consistent with those of A. chabaudi. Moreover, Angiostrongylus-like first stage larvae (L1) were present in the faeces of the animal that was negative for any other cardio-pulmonary parasite. Genetic examination of adult parasites and L1 confirmed the morphological identification as A. chabaudi. Histopathological examination of the lungs showed severe, multifocal to coalescing, chronic, interstitial granulomatous pneumonia due to the presence of adult parasites, larvae and eggs. These findings demonstrate for the first unequivocal time that this nematode reproduces in the European wildcat which should be ultimately considered a definitive host of A. chabaudi. Finally, the L1 of A. chabaudi are described here for the first time, opening new prospects for further studies on this neglected parasite.

The effect of the stage of the ovarian cycle (anoestrus or dioestrus) and of pregnancy on the incidence of gastro-oesophageal reflux in dogs undergoing ovariohysterectomy.

OBJECTIVE: To investigate the potential association of increased blood progesterone (P4 ) concentrations and/or late pregnancy with the incidence of gastro-oesophageal reflux (GOR), in healthy bitches undergoing ovariohysterectomy under general anaesthesia during anoestrus or dioestrus or during the second half of pregnancy. STUDY DESIGN: Prospective observational study. ANIMALS: Ninety-four healthy, female, dogs, aged 1-8 years presented for elective ovariohysterectomy. METHODS: Non-pregnant animals were classified into group A (anoestrus) (n = 35) if blood P4 concentration was sufficiently low or group D (dioestrus) (n = 26) if blood P4 concentration was sufficiently high. All animals in the second half of pregnancy were classified into group P (n = 33). Acepromazine (0.05 mg kg(-1) ) was administered intramuscularly as preanaesthetic medication, and sodium thiopental (10 mg kg(-1) , with additional doses if needed) was administered intravenously (IV) for induction of anaesthesia. After endotracheal intubation, halothane (1.1-1.3% end-tidal concentration) in oxygen was used for maintenance of anaesthesia. Lower oesophageal pH was monitored continuously throughout surgery using a pH-measuring probe. Reflux was considered to have occurred whenever pH values of >7.5 (alkaline reflux) or <4 (acid reflux) were recorded. On completion of surgery, carprofen (4 mg kg(-1) ) was administered IV. Further administration of analgesics post-operatively was dictated by visual analogue scale pain scoring. RESULT: Acid GOR was observed in five of 26 dogs in group D, six of 35 group A, and 12 of 33 group P (p = 0.152). The incidence of GOR in group P approached statistical significance and was higher than the incidence in the combined group A + D (one sided p = 0.044, two sided p = 0.077). CONCLUSIONS: In dogs undergoing ovariohysterectomy, GOR during anaesthesia occurs with a high incidence in dogs in the second half of pregnancy compared to non-pregnant animals during anoestrus or dioestrus. CLINICAL RELEVANCE: Measures could be taken in such cases to avoid the consequences of potential reflux.

A diagnostic model for histologic damage in undescended testes based on testis rigidity measurement: an experimental study with a novel device.

BACKGROUND: We aimed to test whether testis rigidity (hardness) measured using a newly-designed device we previously introduced would offer more reliable assessment of histologic damage in undescended testes than conventional methods (consistency feel at palpation, volume measurement). MATERIALS AND METHODS: Forty-five 18-d-old Lewis rats underwent surgical inhibition of descent of left testes and were followed to 40 (n = 16), 63 (n = 14), or 90 days (n = 15). Another 45 18-d-old Lewis rats were sham operated (left side) and followed likewise (n = 14, n = 15, and n = 16). At the designated time points, testes were exposed bilaterally, rigidity was measured, and consistency at palpation was scored; testes were removed and subjected to length, width, weight measurements, volume calculation, and histomorphometry (mean Johnsen score [MJS], mean tubular diameter [MTD], and mean capsule width [MCW]). Testes of experimental group were compared with ipsilateral testes of sham-operated rats. RESULTS: At all time points, undescended testes had decreased rigidity, MJS, and MTD, increased MCW, decreased volume and weight; contralateral testes remained unaffected. Rigidity was associated only with MJS and MTD, and most strongly with MJS (multiple stepwise linear regression, F = 694.44, P < 0.0005). MJS could be precisely predicted from rigidity: MJS = 0.699 × testis rigidity (F = 1358.82, P < 0.0005). This model showed good fit between predicted and actual MJS values (R(2) = 0.94), low error, nonsignificant bias, sensitivity 75% and specificity 90%. Model validation showed low prediction error and nonsignificant bias, indicating generalizability. Testis volume and palpation proved imprecise MJS predictors. CONCLUSIONS: Testis rigidity is an effective predictor of histologic damage in rat undescended testes, with diagnostic value superior to testis palpation scoring and volume measurement.

Cutaneous Canine Mast Cell Tumor: The Use of Proliferative Markers (Ki-67 and Ki-67 × AgNOR) in Cytological Samples for Diagnosis and Prognosis.

A cytological grading system for canine mast cell tumors (MCTs) has been developed, but its integration into clinical routine has been hindered due to its diagnostic limitations. The aim of this study was to assess the prognostic value of Ki-67 and argyrophilic nucleolar organizing region (AgNOR) markers in cytological MCT samples and to determine cut-off values for these markers in correlation with histopathological grading. Cytological samples were collected prior to surgical excision, and histopathological samples were obtained postsurgery from 45 dogs diagnosed with cutaneous mast cell tumors (MCTs). The cytological specimens were classified using a two-tier grading system, and their Ki-67 (average immunopositive nuclei per 100 cells) and AgNOR (average AgNOR counts per 100 nuclei) signaling was assessed. Through receiver operating characteristic (ROC) analysis, cut-off values for Ki-67 and Ki-67 × AgNOR were determined to better align with histopathological grading (classified as low or high grade according to Kiupel's scoring system). Without the inclusion of proliferative markers, there was a 73% agreement between cytological and histopathological grading. The prediction of histopathological grade was slightly more accurate when assessing Ki-67 and Ki-67 × AgNOR signaling in cytological specimens (75% and 80%, respectively) compared to the initial cytological grading. The cytological assessment of canine MCTs proves beneficial for the initial evaluation, and the incorporation of the evaluation of Ki-67 and AgNOR markers may assist in identifying diagnostically highly malignant MCTs.

Surgical management of squamous cell carcinoma of the canine male external genitalia: 15 cases (1994-2020).

Fifteen male dogs with squamous cell carcinoma of the external genitalia were admitted for further investigation and surgical management between 1994 and 2020. The dogs belonged to various breeds. Thirteen dogs were intact and two were castrated with a median age of 8 years and a median weight of 28 kg. Seven dogs were white-coated and eight nonwhite coated. Scrotal ablation and orchiectomy were performed in four dogs, partial penile amputation in two, partial penile amputation plus partial preputial ablation in one, penile amputation, and scrotal urethrostomy in seven, and local preputial excision in one dog. Postoperative complications included hemorrhage in 10 dogs, bruising at the urethrostomy site in seven, and urethrostomy dehiscence in one dog. Tumor recurrence was recorded in six dogs. Dogs with poorly differentiated tumors that had tumor recurrence had shorter survival and worse prognosis compared to those with well and moderately differentiated tumors. The mean survival time was 48.132 months. After a median follow-up of 23 months (range: 8 to 72 months), eight dogs were alive, five were euthanized and two dogs died from unrelated causes. Surgical excision seems to be a treatment option for dogs with squamous cell carcinoma of the external genitalia.

Can computerized rumen mucosal colorimetry serve as an effective field test for managing subacute ruminal acidosis in feedlot cattle?

Subacute ruminal acidosis (SARA) is one of the major nutritional disorders in the dairy and beef industries, leading to significant financial losses. Diagnosing SARA is challenging due to the need to evaluate multiple parameters, such as milk fat/protein ratio, ruminal lactate, and hemogram, instead of relying on a single definitive symptom or diagnostic method. This study aimed to evaluate the effectiveness of computerized rumen colorimetry in detecting SARA in beef cattle. Over one year, 75 cattle aged 8-10 months from five farms were periodically monitored for rumen pH prior to slaughter. Samples of rumen wall and rumen content were obtained at slaughter for analysis. The study found a positive correlation coefficient between rumen pH and color components, particularly for Red (0.853) and color lightness (L) (0.862). The darkening of the rumen epithelium's color was attributed to the effect of rumen pH on the keratinized layer of the epithelium. Furthermore, an increase in the thickness of ruminal epithelium layers, particularly non-keratinized and total epithelium, was observed in animals with a history of SARA. It is possible that the lower rumen pH increases the rate of replacement of the keratinized epithelium, and the non-keratinized epithelium overgrows to compensate for the need to of produce keratinized layers. In conclusion, computerized rumen colorimetry shows promise as a reliable method for managing SARA in bovine farms by monitoring the condition in the slaughterhouse. Further research is needed to evaluate its effectiveness in detecting SARA in live animals.

Effects of a Curcumin/Silymarin/Yeast-Based Mycotoxin Detoxifier on Redox Status and Growth Performance of Weaned Piglets under Field Conditions.

The aim of this in vivo study was to investigate the effects of a novel mycotoxin detoxifier whose formulation includes clay (bentonite and sepiolite), phytogenic feed additives (curcumin and silymarin) and postbiotics (yeast products) on the health, performance and redox status of weaned piglets under the dietary challenge of fumonisins (FUMs). The study was conducted in duplicate in the course of two independent trials on two different farms. One hundred and fifty (150) weaned piglets per trial farm were allocated into two separate groups: (a) T1 (control group): 75 weaned piglets received FUM-contaminated feed and (b) T2 (experimental group): 75 weaned piglets received FUM-contaminated feed with the mycotoxin-detoxifying agent from the day of weaning (28 days) until 70 days of age. Thiobarbituric acid reactive substances (TBARSs), protein carbonyls (CARBs) and the overall antioxidant capacity (TAC) were assessed in plasma as indicators of redox status at 45 and 70 days of age. Furthermore, mortality and performance parameters were recorded at 28, 45 and 70 days of age, while histopathological examination was performed at the end of the trial period (day 70). The results of the present study reveal the beneficial effects of supplementing a novel mycotoxin detoxifier in the diets of weaners, including improved redox status, potential hepatoprotective properties and enhanced growth performance.

Use of a semitendinosus myocutaneous flap for the coverage of hindlimb full-thickness skin defects in cats.

BACKGROUND: The management of feline hindlimb full-thickness skin defects is challenging. On the other hand, the use of a semitendinosus (ST) myocutaneous flap for their coverage has not been reported. OBJECTIVES: To describe the ST flap and compare it with second intention healing for managing hindlimb full-thickness skin defects. METHODS: In 12 purpose-bred laboratory domestic short-haired cats, two wounds were made on each tibia. The wounds in group A (n = 12) were covered with ST flaps, and those in group B (n = 12) were left to heal by second intention. In both groups, clinical assessment scoring and planimetry were performed between one-30 d postoperatively. Computed tomography-angiography (CTA) was performed on days zero, 10, and 30, and histological examinations were performed on days zero and 14 and at 6 and 12 mon postoperatively. RESULTS: Statistically significant differences in the clinical assessment scores were observed between groups A and B on days 14 (p = 0.046) and 21 (p = 0.016). On the other hand, the time for complete healing was similar in the two groups. CTA revealed significant differences in the muscle width (day 0 compared to days 10 and 30 [p = 0.001, p = 0.026, respectively], and days 10 to 30 [p = 0.022]), ST muscle density, and the caliber of the distal caudal femoral artery and vein (day 0 compared to day 10 [p < 0.001], and days 10 to 30 [p < 0.001]). Histologically significant differences in inflammation, degeneration, edema, neovascularization, and fibrosis were observed on day 14 compared to zero and 6 mon, but no differences were found between the time interval of 6 and 12 mon. CONCLUSIONS: An ST flap can be used effectively to manage hindlimb full-thickness skin defects.

Evaluation of Survival of Subdermal Plexus Skin Flaps in 8 Cats After Injection of Platelet Rich Plasma.

The aim of the present study was to evaluate the effect of platelet-rich plasma injection on the survival of experimentally developed subdermal plexus skin flaps in cats. Two flaps, 2 cm wide and 6 cm long were created bilaterally in the dorsal midline in 8 cats. Each flap was randomized to 1 of 2 groups: platelet-rich plasma injection and control. After flap development, the flaps were immediately placed back on the recipient bed. Then 1.8 mL of platelet-rich plasma was equally injected into 6 different parts of the treatment flap. All flaps were evaluated macroscopically daily and on days 0, 7, 14, and 25 by planimetry, Laser Doppler flowmetry, and histology. Mean flap survival on day 14 was 80.437 % (±22.745) for the treatment group and 66.516 % (±24,12) for the control group with no statistically significant difference between the 2 groups (P = .158). Histologically, a significant difference was found between the base of the PRP and the control flap in edema score on day 25 (P = .034). In conclusion, there is no evidence to support the use of platelet-rich plasma in subdermal plexus flaps in cats. However, the use of platelet-rich plasma may aid in reducing edema of subdermal plexus flaps.

Comparison of Two Surgical Techniques Based on the Semitendinosus Myocutaneous Flap in Cats.

The objective of this experimental study was to compare the semitendinosus (ST) to the split-semitendinosus (SST) myocutaneous flap in covering distal limb skin defects in cats. Twenty-eight purpose-bred laboratory DSH cats were used and allocated into two groups (ST-group (A); n = 14, SST-group (B); n = 14). ST flaps, based on the distal muscle pedicle, and SST flaps, after longitudinal division of the muscle based on both muscle pedicles, were tested over skin defects created on the medial distal tibia. Clinical assessment, planimetry, CT-angiography (CTA) and histological examination were compared between groups. Days to complete flap healing between ST and SST-flaps (30.36 ± 9.1, 32.29 ± 5.44, respectively) and final total flap areas (68.36% ± 27.18, 51.83% ± 22.48, respectively) revealed no significant differences. On CTAs, the caliber of the distal caudal femoral vein on day 10 was statistically significant higher (p < 0.001) for group A and a significantly higher caliber of the distal caudal femoral artery on day 30 for group B (p = 0.021). Histology revealed statistically higher degeneration at 6 months (p = 0.047) for group A, and statistically higher fibrosis at 12 months (p = 0.019) for group B. Both ST and SST flaps had similar healing times and provided coverage of skin tibial defects in cats.

Histopathological Pulmonary Lesions in 1st-Day Newborn Piglets Derived from PRRSV-1 MLV Vaccinated Sows at the Last Stage of Gestation.

Modified live virus (MLV) vaccines for the control of porcine respiratory and reproductive syndrome virus (PRRSV) have been associated with the vertical and horizontal transmission of vaccine viruses. The present study aimed to describe pathological lung lesions in piglets born by gilts vaccinated with PRRSV-1 MLV. In total, 25 gilts were vaccinated at late gestation (100th day) and were divided into five groups according to the different vaccines (Vac) used: no vaccine-control group, Vac-1-strain DV, Vac-2-strain VP-046 BIS, Vac-3-strain 94881, Vac-4-strain 96V198. Within the first 0-9 h of the farrowing, blood samples were collected from all newborn piglets and lung samples were exanimated grossly, histopathologically and with scanning electron microscopy. PRRSV (RT-PCR-positive) and antibodies were detected in the serum of piglets from gilts vaccinated with Vac-2. In these piglets, moderate to severe interstitial pneumonia with thickened alveolar septa was noticed. Type II pneumocyte hyperplasia was also observed. The rest of the trial piglets showed unremarkable lung lesions. Phylogenetic analysis revealed the 98.7% similarity of the PRRSV field strain (GR 2019-1) to the PRRS MLV vaccine strain VP-046 BIS. In conclusion, the Vac-2 PRRSV vaccine strain can act as an infectious strain when vaccination is administrated at late gestation, causing lung lesions.

SGCD Missense Variant in a Lagotto Romagnolo Dog with Autosomal Recessively Inherited Limb-Girdle Muscular Dystrophy.

An 8-month-old female Lagotto Romagnolo dog was presented for a 1-month history of an initial severe reluctance to move, rapidly progressing to a marked stiff gait and progressive muscular weakness and evolving to tetraparesis, which persuaded the owner to request euthanasia. A primary muscle pathology was supported by necropsy and histopathological findings. Macroscopically, the muscles were moderately atrophic, except for the diaphragm and the neck muscles, which were markedly thickened. Histologically, all the skeletal muscles examined showed atrophy, hypertrophy, necrosis with calcification of the fibers, and mild fibrosis and inflammation. On immunohistochemistry, all three dystrophin domains and sarcoglycan proteins were absent. On Western blot analysis, no band was present for delta sarcoglycan. We sequenced the genome of the affected dog and compared the data to more than 900 control genomes of different dog breeds. Genetic analysis revealed a homozygous private protein-changing variant in the SGCD gene encoding delta- sarcoglycan in the affected dog. The variant was predicted to induce a SGCD:p.(Leu242Pro) change in the protein. In silico tools predicted the change to be deleterious. Other 770 Lagotto Romagnolo dogs were genotyped for the variant and all found to be homozygous wild type. Based on current knowledge of gene function in other mammalian species, including humans, hamsters, and dogs, we propose the SGCD missense variant as the causative variant of the observed form of muscular dystrophy in the index case. The absence of the variant allele in the Lagotto Romagnolo breeding population indicates a rare allele that has appeared recently.

The Use of Stromal Vascular Fraction in Long Bone Defect Healing in Sheep.

The objectives of the present study were to evaluate (a) the feasibility of using stromal vascular fraction (SVF) and nanocrystalline hydroxyapatite (nHA) paste in combination for the treatment of segmental bone defect, (b) the quality of the callus produced, (c) the potential improvement of the autograft technique, and (d) the direct comparison of the biomaterial to the use of autogenous cancellous bone. Unilateral, segmental mid-diaphyseal bone defect was created on the right metatarsus of skeletally mature sheep animals (n = 24) under anesthesia (D0). Residual segments were stabilized by stainless-steel plates and appropriate screws. Defects were managed as follows: group A: use of nHA paste to filling, group B: use of autogenous bone graft mixed with nHA bone paste, placed in defect, group C: use of SVF mixed with nHA bone paste injected into defect, group D: use of bone graft and SVF with nHA paste before apposition in bone defect. SVF had been previously isolated from adipose tissue of the animals intra-operatively after digestion with collagenase solution and neutralization. Animals were evaluated clinically and by X-raying and ultrasonographic examination of the defect, at regular intervals, until D90. Ultrasonographic assessment performed along the length of the defect included calculation of the length of the bone defect and assessment of vascularization. SVF was successfully isolated from group C and D animals, with the average yield being 1.77 × 106 cells. The comparison of clinical scores (based on the 'Kaler scale') on each post-operative day indicated significant differences between the four groups on D1 to D30 (p < 0.01); the median clinical score within group A was 2.5 for D1-D30 and 1 for the entire period; respective scores for other groups were 1.5 (p = 0.07) and 0 (p = 0.033). Differences in radiographic assessment scores were significant for scores obtained on D60 (p = 0.049) and D90 (p = 0.006). There was a significant difference between the four groups in the length of the bone defect, as assessed ultrasonographically, for the entire length of the study; median values were 8, 8.5, 6, and 8 mm for groups A, B, C, and D, respectively (p = 0.008). There was a significance in the differences between median scores obtained during the histopathological examination: 2, 11, 13.5, and 12 for group A, B, C, and D (p = 0.022). There was an inverse correlation between the overall scores of histopathological evaluations and the length of the bone defect (observed on D90) (p < 0.0001) and a correlation between the overall scores and the radiographic assessment scores (obtained on D90) (p < 0.0001). This is the first study in which the efficacy of fresh autologous Stromal Vascular Fraction (SVF) from adipose tissue in enhancing bone healing in a long, weight-bearing, diaphyseal bone was evaluated. It is concluded that the lumbosacral region was an attractive site for harvesting adipose tissue, the use of SVF contributed to faster rehabilitation post-operatively, and SVF significantly enhanced bone formation; in general, the results indicated an osteogenic potential of SVF comparable to the gold standard autologous bone graft.

Beneficiary Effect of Fibrin Glue on Healing of Ileoileal Anastomoses in Rats.

BACKGROUND/AIM: Intestinal anastomosis' integrity is crucial in surgery. This study aimed to investigate whether fibrin glue (FG) (a fibrin sealant containing human factor XII and fibrinogen) has a positive effect on the healing and the integrity of the ileoileal anastomosis in rats. MATERIALS AND METHODS: Twenty Wistar rats underwent enterotomy, ileoileal anastomosis and divided into four groups (A: complete anastomosis-no FG, B: complete anastomosis-FG, C: incomplete anastomosis-no FG, D: incomplete anastomosis-FG). Data included leak, adhesions, bursting pressure of the anastomosis, neoangiogenesis, and hydroxyproline levels. RESULTS: Angiogenesis was significantly higher in group B compared to group A (p=0.019). There were no significant differences between groups A and B regarding adhesions, hydroxyproline, and bursting pressure (p=0.500, p=0.158 and p=0.829, respectively). Hydroxyproline levels were higher in group D compared to C, but did not reach significance (p=0.098). CONCLUSION: Fibrin glue has a positive effect on ileoileal anastomoses. It is not entirely clear whether this effect is due to mechanical support or to the facilitation of the healing process or both. Further research is needed before FG can be applied to humans.

Τhe Effect of Opioid Administration on Cytologic and Histopathologic Diagnosis of Canine Cutaneous Mast Cell Tumors Treated by Surgical Excision.

Mast cell tumor (MCT) is a frequent cutaneous tumor in dogs, with a variable biological behavior. Studies correlate cytologic and histopathologic features of MCTs with their biological behavior, prognosis, and response to treatment. The use of preoperative opioids is common in canine patients undergoing surgical removal of these tumors. Certain opioids can induce or downregulate mast cell degranulation and influence cancer progression. The aim of the present study was to investigate whether the administration of morphine or butorphanol during surgical excision of canine cutaneous MCTs affects their cytologic and histopathologic appearance, thus influencing cytologic and histopathologic grading. This was a prospective, blinded, randomized, cohort clinical study. Forty-five dogs with cutaneous MCTs were randomly allocated into three groups according to preanaesthetic medication: dexmedetomidine combined with morphine (group M) or butorphanol (group B) or normal saline (group C). Cytologic specimens and histopathologic samples were obtained both prior to and after surgery. Samples were graded according to Kiupel's and Patnaik's systems, examined immunohistochemically for Ki-67 protein (Ki-67) and c-kit proto-oncogene product (KIT) expression, and histochemically for argyrophilic nucleolar organizing regions (AgNORs). Based on both Kiupel's and Patnaik's systems, no statistically significant differences were noted concerning the number of cases with grading discrepancies in grades allocated prior to versus after surgery among the groups. The same applied for cytological grading and immunohistochemical and histochemical evaluation. It seems that administration of morphine or butorphanol as part of the preanesthetic medication for surgical removal of canine cutaneous mast cell tumors does not influence histopathologic and cytologic grading of MCTs.