Platelet-rich plasma attenuates the UPEC-induced cystitis via inhibiting MMP-2,9 activities and downregulation of NGF and VEGF in Canis Lupus Familiaris model.

One of the most prevalent disorders of the urinary system is urinary tract infection, which is mostly brought on by uropathogenic Escherichia coli (UPEC). The objective of this study was to evaluate the regenerative therapeutic and antibacterial efficacy of PRP for induced bacterial cystitis in dogs in comparison to conventional antibiotics. 25 healthy male mongrel dogs were divided into 5 groups (n = 5). Control negative group that received neither induced infection nor treatments. 20 dogs were randomized into 4 groups after two weeks of induction of UPEC cystitis into; Group 1 (control positive; G1) received weekly intravesicular instillation of sodium chloride 0.9%. Group 2 (syst/PRP; G2), treated with both systemic intramuscular antibiotic and weekly intravesicular instillation of PRP; Group 3 (PRP; G3), treated with weekly intravesicular instillation of PRP, and Group 4 (syst; G4) treated with an intramuscular systemic antibiotic. Animals were subjected to weekly clinical, ultrasonographic evaluation, urinary microbiological analysis, and redox status biomarkers estimation. Urinary matrix metalloproteinases (MMP-2, MMP-9) and urinary gene expression for platelet-derived growth factor -B (PDGF-B), nerve growth factor (NGF), and vascular endothelial growth factor (VEGF) were measured. At the end of the study, dogs were euthanized, and the bladder tissues were examined macroscopically, histologically, and immunohistochemically for NF-κB P65 and Cox-2. The PRP-treated group showed significant improvement for all the clinical, Doppler parameters, and the urinary redox status (p < 0.05). The urinary MMPs activity was significantly decreased in the PRP-treated group and the expression level of urinary NGF and VEGF were downregulated while PDGFB was significantly upregulated (p < 0.05). Meanwhile, the urinary viable cell count was significantly reduced in all treatments (P < 0.05). Gross examination of bladder tissue showed marked improvement for the PRP-treated group, expressed in the histopathological findings. Immunohistochemical analysis revealed a marked increase in Cox-2 and NF-κB P65 in the PRP-treated group (P < 0.05). autologous CaCl2-activated PRP was able to overcome the bacterial infection, generating an inflammatory environment to overcome the old one and initiate tissue healing. Hence, PRP is a promising alternative therapeutic for UPEC cystitis instead of conventional antibiotics.

Effect of pyometra on vascularity alterations, oxidative stress, histopathology and inflammatory molecules in feline.

This study aimed to investigate blood flow, hemodynamical features by Doppler ultrasound, the oxidative stress biomarkers from serum samples, and histopathology from uterine tissue, in healthy queens and queens with pyometra. Twenty queens were categorized into two groups, according to signs, history, and ultrasound findings, as pyometra and control healthy queens. Doppler ultrasonography, total antioxidant capacity (TAC), malondialdehyde (MDA), albumin, bacteriological isolation, histopathology, and immunohistochemistry of tumor necrosis factor-alpha (TNF-α) and nuclear factor kappa B (NF-κB) P65 were performed. Uterine diameter and thickness increased significantly in the pyometra group compared to control. Uterine peak velocity and flow rate were significantly higher in the control group. The pyometra group showed a significant decrease in albumin, TAC, and a significant increase in MDA. Fibrosis and mononuclear inflammatory cell infiltration were seen in the pyometra samples. The mean area percentage of TNF-α expression in the uteri of the pyometra group was higher. The expression of NF-κB P65 in the uteri in the pyometra group was significantly higher. Doppler ultrasonography can provide valuable information for diagnosing pyometra in queens by elevating the uterine thickness with reducing blood flow rate. Oxidative stress, TNF-α, and NF-κB expression alterations varied between pyometra and control groups.

The therapeutic potential of Camel Wharton jelly mesenchymal stem cells (CWJ-MSCs) in canine chronic kidney disease model.

BACKGROUND: Chronic kidney disease (CKD) is a worldwide health problem that its incidence increases nowadays with the increase in the risk of environmental pollution. CKD can progress to end-stage renal disease (ESRD) which usually ends fatally. This study aimed to examine the therapeutic potential of Camel Wharton jelly-mesenchymal stem cells (CWJ-MSCs) in chronic kidney disease model induced in dogs. METHODS: CWJ-MSCs were injected directed to the kidney with ultrasonographic guidance in dogs with 5/6 nephrectomy to evaluate its therapeutic potency in such cases. Analysis of variance was applied in normally distributed quantitative variables while a non-parametric Mann-Whitney test was used for non-normally distributed quantitative variables. RESULTS: The serum urea and creatinine in the treated group were significantly decreased transferring dogs in the treated group from stage 3 to stage 2 CKD according to the IRIS staging system. Histopathology of renal tissue revealed improving CKD lesions by increasing regeneration of degenerated tubules, maintaining the integrity of glomeruli. New vascularization with blood vessels remodeling were common findings. Periodic acid Schiff stain of renal tissue showed the integrity of renal tubules and thickness of the glomerular basement membrane. Fibrosis of cortex and medulla was lower in the treated group than in the CKD model as monitored by Mallory's trichrome stain (MTC). NGAL and KIM-1 genes expression were decreased while VEGF and EGF genes expression were increased indicating renal tissue repair. CONCLUSIONS: CWJ-MSCs have a therapeutic potential in the CKD model induced in dogs.

The efficiency of intrauterine infusion of platelet-rich plasma in the treatment of acute endometritis as assessed by endoscopic, Doppler, oxidative, immunohistochemical, and gene expression alterations in jennies.

This study used autologous platelet-rich plasma (PRP) to treat acute endometritis in jennies with follow-up for alterations in uterine hemodynamics, endoscopic, immunohistochemistry, oxidant/antioxidant imbalance, pro-inflammatory regulatory molecules, and transmembrane mucin expressions. Ten jennies suffering from endometritis (acute type; n = 10) were included in the study. PRP was prepared from each animal and two intrauterine infusions one week apart were administrated. Examination and follow-up were done physically, ultrasonographically, endoscopically and samples were taken for histopathology, immunohistochemistry, and bacteriological examination. Blood and uterine fluid samples were taken to estimate biochemical and oxidative stress alterations. Expression of TRAF6 and MUC1 genes was investigated in uterine fluid, at days -1 (day of diagnosis establishment), 7, 14, and 21. Uterine bacteriological examination showed a decrease in bacterial isolates after PRP treatment. The uterine thickness and uterine vascular perfusion as illustrated by color Doppler ultrasonography were significantly decreased in jennies treated by PRP. Uterine spectral wave pattern showed a significant linear increase in pulsatility index only. Three weeks after first PRP treatment, white light endoscopic examination revealed normal uterine body mucosa and uterine horn folds. A high nuclear factor (NF-κB) expression was seen in the mononuclear cells. A significant reduction in oxidative stress biomarkers in both serum and uterine fluid was recorded after PRP treatment. The TRAF-1 gene expression significantly decreased gradually after intrauterine PRP infusion. The MUC-1 gene expression significantly decreased gradually after intrauterine PRP infusion. Both genes were within normal levels by week 3. Endometritis in jennies is associated with an oxidative process, alterations in serum biochemical parameters, Doppler indices, endoscopic appearance, high NF-κB expression, and upregulation of TRAF-1 and MUC-1 expressions. Two intrauterine infusions of autologous PRP restored normal endometrial appearance after acute endometritis.

MicroRNA-21 expression, serum tumor markers, and immunohistochemistry in canine mammary tumors.

BACKGROUND: Canine mammary tumors (CMTs) are one of the most common malignancies in dogs and are associated with significant mortality. Serum tumor markers and non-coding microRNAs have gained widespread popularity in human oncology studies. The present study has two aims, first one is to investigate the miR-21 expression compared with changes in serum tumor markers (CEA and CA15-3) in CMT. The second aim is to detect the immunohistochemistry markers as vimentin, P63, and -SMA in CMT. METHODS: This study enrolled 17 female dogs: 10 with mammary tumors and seven controls without tumors. Blood samples were collected to measure miR-21, CEA, and CA 15-3, and histological samples were prepared for histological grading and immunohistochemistry. RESULTS: CA 15-3 was elevated in all animals, whereas CEA levels showed no change compared with controls. miR-21 was upregulated 12.84-fold in animals with CMT. The most frequently recorded CMT was the mixed type. Myoepithelial cells were identified by P63 immunoreactivity, but not SMA. High expression of miR-21 was observed with positive vimentin immunoreactivity, indicating the mesenchymal origin of the tumor cells. CONCLUSION: The present study showed that miR-21 was elevated to a greater extent than CA 15-3 (12.84-fold vs. threefold). Tumors that was positive for vimentin immunoreactivity was also associated with an elevation in the levels of miR-21, showing that miR-21 is released from mesenchymal cells. These findings support the hypothesis that miR-21 may be a more sensitive, noninvasive indicator for CMT.

The other side to the use of active targeting ligands; the case of folic acid in the targeting of breast cancer.

Due to its overexpression in cancer cells, the folate receptor (FR) is heavily exploited in the active targeting of nanoparticles (NPs). Its ligand, folic acid (FA) is as a consequence widely used as a NP targeting ligand. Although rather popular and successful in principle, recent data has shown that FA may result in breast cancer initiation and progression, which questions the suitability of FA as NP cancer targeting ligand. In this work, intravenous administration of free FA to healthy female mice resulted in breast tissue dysplasia, hyperplasia and in the increased expression of human epidermal growth factor receptor-2 (HER2), folate receptor (FR), cancer antigen 15-3 (CA15.3), vascular endothelial growth factor (VEGF), signal transducer and activator of transcription 3 (STAT3) and the pro-inflammatory cytokines, tumor necrosis factor alpha (TNFα), interleukin-6 (IL-6) and interleukin-1ß. In addition to the reduction in IL2. To evaluate the suitability and safety of FA as NP targeting ligand in breast cancer, small (≈ 150 nm) and large (≈ 500 nm) chitosan NPs were formulated and decorated with two densities of FA. The success of active targeting by FA was confirmed in two breast cancer cell lines (MCF-7 and MDA-MB-231 cells) in comparison to HEK293 cells. FA modified NPs that demonstrated successful active targeting in-vitro were assessed in-vivo. Upon intravenous administration, large NPs modified with a high density of FA accumulated in the breast tissue and resulted in similar effects as those observed with free FA. These results therefore question the suitability of FA as a targeting ligand in breast cancer and shed light on the importance of considering the activity (other than targeting) of the ligands used in NP active targeting.

Implanted subcutaneous versus intraperitoneal bioscaffold seeded with hepatocyte-like cells: functional evaluation.

BACKGROUND AND OBJECTIVES: The X-linked bleeding disorder, hemophilia A, is caused by defective production of factor VIII (FVIII). Hemophilic patients require regular FVIII infusions. Recombinant factor replacement poses the safest line of therapy. However, its main drawbacks are high expenses and the higher liability for formation of inhibitors. Recent studies confirmed the ability of bone marrow-derived stem cells to secrete FVIII. This study aims to generate bioscaffold from decellularized liver and subsequently seed it with trans-differentiated human stem cells into hepatic-like cells. This scaffold can then be implanted intraperitoneally or subcutaneously to provide FVIII. METHODS: After generation of the bioscaffold, seeding of discoid scaffolds with trans-differentiated human hepatocyte-like cells was performed. Then, the generated organoid was implanted into peritoneal cavity or subcutaneous tissue of experimental rats. RESULTS: Serum human FVIII was significantly increased in rats subjected to subcutaneous implantation compared intraperitoneal implantation. Immunostaining for detecting Cytokeratin 19 and human anti-globulin confirmed the presence of mature human hepatocytes that were significantly increased in subcutaneous implanted scaffold compared to the intraperitoneal one. CONCLUSION: Implantation of decellularized bioscaffold seeded with trans-differentiated stem cells in rats was successful to establish production of FVIII. Subcutaneous implantation showed higher FVIII levels than intraperitoneal implantation.

The implication of autoantibodies in early diagnosis and monitoring of plasmonic photothermal therapy in the treatment of feline mammary carcinoma.

Feline mammary carcinoma (FMC) shows great similarities to human breast cancer in the cellular and molecular levels. So, in cats as in humans, the role of immune responses is indicated to detect and follow up the development of tumors. As a new breast cancer therapeutic approach, Plasmonic Photothermal Therapy (PPTT) is an effective localized treatment for canine and feline mammary-carcinoma. Its systemic effect has not been inquired yet and needs many studies to hypothesis how the PPTT eradicates tumor cells. In this study, it is the first time to detect (P53, PCNA, MUC-1, and C-MYC) feline autoantibodies (AAbs), study the relationship between PCNA AAbs and mammary-tumors, and investigate the effect of PPTT on the humoral immune response of cats with mammary-carcinoma through detection of AAbs level before, during, and after the treatment. The four-AAbs panel was evaluated in serum of normal and clinically diagnosed cats with mammary tumors using Enzyme-Linked Immunosorbent Assay. The panel showed 100% specificity and 93.7% sensitivity to mammary tumors. The panel was evaluated in PPTT monotherapy, mastectomy monotherapy, and combination therapy. PPTT monotherapy decreased AAbs level significantly while mastectomy monotherapy and combination therapy had a nonsignificant effect on AAbs level.

Corneal Ulcer in Dogs and Cats: Novel Clinical Application of Regenerative Therapy Using Subconjunctival Injection of Autologous Platelet-Rich Plasma.

Background: Corneal ulcer could be a major source of distress in small animals, with many contributing agents. In recent years, few studies evaluated the efficacy of platelet-rich plasma (PRP) in healing corneal ulcers. Aim: This study aimed to assess the ability of subconjunctival injection of autologous PRP in the treatment of corneal ulcers in dogs and cats as well as estimate the expression of matrix metalloproteinase (MMP)-2, MMP-9, and oxidative stress biomarkers in these patients. Methods: A total number of 28 animals (16 cats and 12 dogs) were enrolled in this study. Each animal was subjected to clinical, neurologic, and ophthalmic examinations where the type of ulcer was documented. Tear samples were collected for evaluation of oxidative biomarkers and MMPs; conjunctival swabs were taken to identify the involved organism. PRP was prepared from each animal and given as subconjunctival injection; numbers of injections were done according to case response. Clinical follow-up was done and documented for each case. Results: In cat patients, female and Persian cats were most affected; unilateral and superficial ulcers were most recorded. In male dogs, unilateral, and superficial ulcers were most recorded. FHV-1 was most identified in cats, while Staphylococcus aureus was most identified in dogs. Numbers of injections needed to achieve healing were recorded, with 50% of dogs needing two injections with 1-week intervals and 50% of cats needed three injections with 1-week intervals. Alterations in both oxidative biomarkers and MMPs were recorded in affected animals. Conclusion: The use of autologous PRP as a subconjunctival injection in treating corneal ulcers in dogs and cats is effective. The number of injections is the case and corneal ulcer type-dependent. Clinical Significance: Autologous PRP as a subconjunctival injection in treating corneal ulcer is a relatively cheap, safe method and can be done in the clinical setting.

Uterine hemodynamic patterns, oxidative stress, and chromoendoscopy in mares with endometritis.

This study aimed to evaluate the use of Doppler ultrasound for determining alterations in uterine thickness and uterine arteries, use of chromohysteroscopy in evaluating endometritis with biopsies, and the impact of endometritis on oxidative and hormonal status. Twenty Arabian mares were divided into control and endometritis groups. Each underwent a physical examination, Doppler ultrasound, chromoendoscopy, and uterine biopsy. Blood samples from each animal were collected for hematologic, hormonal, and oxidative analysis. Results revealed a significant increase in estradiol, malondialdehyde (MDA), and nitric oxide metabolites (NOMs) associated with a significant decrease in progesterone and total antioxidant capacity (TAC) in the endometritis group. There was a significant difference in uterine thickness in the endometritis group. Uterine blood flow analyzed by color mode showed a significant increase in the endometritis group, while the spectral mode showed a significant increase in Doppler velocities and blood flow rate accompanied by a significant decrease in Doppler indices. The endometritis group showed ridged, bluish color in the endometrial wall, indicating hyperemia. Biopsy showed mononuclear inflammatory cells and edema in the endometrial interstitial tissue with sloughing of the epithelial lining. Conclusion: Endometritis in mares is associated with oxidative processes and presents with an increase in MDA and NOMs and a decrease in TAC. Doppler ultrasound improved endometritis diagnosis because it showed alterations in uterine vascularization. Chromoendoscopy improved the accuracy in taking biopsy samples because it allows the practitioner to select appropriate areas for biopsy to improve histopathological diagnostic outcomes. Therefore, this technique is worthy of further study in the field of veterinary equine reproduction.

Oesophageal foreign bodies in cats: Clinical and anatomic findings.

BACKGROUND: Anatomical feline models can aid in understanding the relationships between clinical findings and anatomical features and the course of foreign bodies passing through the oesophagus. This study has two goals 1) to assess feline oesophageal foreign bodies in feline patients using physical, radiologic and endoscopic examination and, how their location influences treatment plans and complications. 2) How the anatomical sharp angle of the oesophagus contribute to foreign body lodgement. Thirty-five cats were enrolled in this study; 30 of them were clinically ill, and five cats were used for anatomical study. RESULTS: Cats with clinical signs underwent complete clinical and radiologic examination. Endoscopy was performed in only five cases. The site with the highest occurrence of foreign body lodgement was the oesophageal entrance, caudal to the pharynx (63.3%), followed by the thoracic inlet (26.7%) and the mid-cervical region of the oesophagus (10%). Two types of foreign bodies were identified: sewing needles (25/30) and bone (5/30). Radiography was able to identify the location and nature of the foreign body in all 30 affected cats. Therapeutic regimens were applied according to the nature and location of the foreign body and any associated complications. Removal of the foreign body was achieved using Rochester pean artery forceps in 17/30 cases, using full surgical intervention in 8/30 cases, and during endoscopy in 5/30 cases. CONCLUSION: The results suggest that the location of the foreign body is strongly related to combination of consumed foreign body type and anatomic features of the cat oesophagus. The feline oesophagus has a variety of sharp angles that facilitate the entrapment of rigid linear and angular foreign bodies. Radiographic imaging remains the most frequently used diagnostic modality for determining the lodgement site and nature of radiopaque foreign bodies. Over all complication rate was low (6/30).

Gross Anatomical, Radiographic and Doppler Sonographic Approach to the Infra-auricular Parotid Region in Donkey (Equus asinus).

The present study was performed on 12 heads of donkey's cadavers of both sexes for different anatomic techniques and on 20 live adult donkeys for ultrasonographic approaches of local anesthetic techniques. The aim was to achieve safe desensitization of the ear canal and tympanic membrane in addition to measuring different parameters of the structures occupying the infra-auricular parotid region. The internal auricular nerve was divided into two fine branches and constituting, at its origin, a characteristic V-shape with the caudal auricular nerve. The styloid process of auricular cartilage was an adequate landmark for ultrasonographic needle-guided anesthesia for internal auricular and auriculopalpebral nerves, whereas the great auricular nerve was easily palpated subcutaneously that showed safety and success in all cases. The auricular branch of mandibular nerve joined the auriculopalpebral branch of facial nerve. The maxillary vein was descending, partially embedded within the texture of the parotid glandular tissue. The parotid gland divided into five segments was clearly demarcated by maxillary vein tributaries with three main collecting radicles pouring into the parotid duct. The mandibular duct received about 12-15 fine radicles and supplied with a separate branch from the external carotid artery. Using the Doppler sonographic technique in donkeys for diagnosis of ear affections, evaluation of retrograde sialography to salivary glands with their blood vasculature and their indices might be helpful for the detection of different critical abnormalities, such as stenosis, thrombosis, and other vasopathological affections through measuring their resistivity and pulsatility indices.

Antimicrobial action of autologous platelet-rich plasma on MRSA-infected skin wounds in dogs.

Effective antimicrobial preparations, other than antibiotics, are important for the treatment of potentially fatal drug-resistant infections. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the leading causes of hospital-acquired and post- operative infections. Fortunately, the antimicrobial properties of platelet-rich plasma (PRP) against various microorganisms enable its potential use as an alternative to conventional antibiotics. The present work was designed to evaluate the hypothesized antimicrobial activity of PRP against MRSA infected skin wounds. Six adult male dogs were divided equally into control and PRP groups. Unilateral circular full-thickness skin wounds were created then a MRSA suspension was injected locally. Treatment started at 1st week post infection with subcutaneous infiltration of autologous activated PRP every week in the PRP group and with topical application of clindamycin cream twice daily in the control group. PRP decreased wound size and significantly increased wound contractility and re-epithelization, as confirmed by histopathological and immunohistochemical findings. Also PRP treated group showed significant decrease in ROS and redox imbalance with over expression of the TNF-α and VEGFA genes that indicate angiogenesis and maximum antibacterial activity after three weeks. In conclusion, CaCl2-activated PRP exhibited antimicrobial activity against MRSA infection, which improved the infected wound healing re-epithelization and granulation tissue formation.

Stimulation of Bladder Acupoints by Cloprostenol for Treating Back Soreness in Athletic Horses.

Twenty-five Thoroughbred jumper geldings suffered back soreness with poor performance, and 5 control horses were assessed by archived computer data, clinical examination, and laboratory analyses of complete blood picture, serum enzymes, and cortisol level, before and after cloprostenol-pharmacopuncture. The 25 diseased horses before therapy showed significant increases in aspartate aminotransferase and creatine phosphokinase with clinical pains scored mild in 15 horses, moderate in 9 horses, and severe in one horse, without changes in the hormonal and hematological data. After therapy, they responded by an increase of heart rate (57.8 ± 4.3 bpm), body temperature (38.5 ± 0.7°C), respiration rate (28.3 ± 2.1 bpm), and capillary refilling time (CRT) (1.0 ± 0.0). On the 2nd day, a significant decrease in the mean levels of aspartate aminotransferase and creatine phosphokinase (P = 0.001) was detected, while on the 4th day, they mimed the level of the 5 controls, and on the 6th day, they showed a significant decrease (P = 0.002). The serum cortisol level showed a significant increase on the 6th day of treatment (P = 0.013). The blood picture showed significant increases in red blood cells, mean corpuscular volume, platelets, white blood cells, hemoglobin, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, mean platelet volume, platelet distribution width, lymphocytes, plateletcrit, and large platelet concentration ratio (P < 0.05) and nonsignificant changes in hematocrit, granulocytes, and midocytes. The improved blood parameters, enzymes, hormones, and performance progress after cloprostenol-pharmacopuncture proved its effectiveness in treating back soreness in athletic horses.

Gold Nanorod-Assisted Photothermal Therapy Decreases Bleeding during Breast Cancer Surgery in Dogs and Cats.

For localized tumors, gold nanorod (AuNR)-assisted plasmonic photothermal therapy (PPTT) is a potentially effective alternative to traditional surgery, in which AuNRs absorb near-infrared light and convert it to heat in order to kill cancer cells. However, for large tumors (volume ≥ 20 cm3), an uneven distribution of AuNRs might cause inhomogeneity of the heat distribution inside the tumor. Surgery is frequently recommended for removing large tumors, but it is associated with a high risk of cancer recurrence and metastasis. Here, we applied PPTT before surgery, which showed improved treatment for large tumors. We divided the animals (eight cats/dogs) into two groups: Group I (control), where three cases were solely treated with surgery, laser, or AuNRs alone, resulting in recurrence and metastasis; and Group II, where animals were treated with PPTT before surgery. In Group II, four out of the five cases had tumor regression without any recurrence or metastasis. Interestingly, we observed that applying PPTT before surgery displayed reduced bleeding during tumor removal, supported by histopathology that showed altered blood vessels. In conclusion, our study showed that applying AuNR-assisted PPTT (AuNRs-PPTT) before surgery could significantly affect blood vessels inside the tumor, leading to a decreased amount of bleeding during surgery, which can potentially decrease the risk of metastasis and blood loss during surgery.

Ultrasonography guidance for total splenectomy in donkeys.

There are varieties of surgical approaches reported for equine splenectomy and all of them were dealing with the most reachable situation of splenic hilus and easy handling of the spleen. The aim of this work was to establish the normal ultrasound parameters of spleen in donkeys (normal echogenicity, hilus situation, topographic location and correlation with neighboring organs) as a guide to select the best approach for total splenectomy in donkeys. Splenic ultrasound was carried out on six normal donkeys before experimental total splenectomy in the standing position. The splenic topographic location was recorded among 4 rows including 30 squares. These animals were divided into two groups according to the surgical approach of total splenectomy. Total splenectomy after left 16th and 17th ribs partial resection in standing position was carried out in group1 and group 2, respectively. Ultrasonographically, the spleen had homogenously echogenic pattern and appeared hyperechoic to the liver. Only one third of the spleen was located in front of the 16th rib where the hilus and splenic blood vessels were nearly under the 16th rib. The splenic artery and splenic vein were ultrasonographically visualized between the left 16th and 17th ribs 10-15 cm from dorsal midline. This area was the site of the important ligation during total splenectomy. In conclusion, ultrasonography guidance for total splenectomy in donkeys assisted the surgical findings and proved that technique following partial resection of the 17th rib at the standing position is the most convenient surgical approach for total splenectomy in donkeys.

Evaluation of chromocystoscopy in the diagnosis of cystitis in female donkeys.

Early detection of cystitis in equine is essential to improve the prognosis and outcome of therapy. However, the conventional white light endoscopy is not sufficiently accurate for this purpose. Hence, this study evaluated chromoendoscopy as a recent diagnostic tool for cystitis in female donkeys. For this purpose, 5 apparently normal donkeys (control group) and 5 female donkeys with cystitis (diseased group) were used. Physical and rectal examinations, urine analysis, white light cystoscopy, methylene blue-based chromoendoscopy and histopathology were performed in all animals. Turbid urine exhibiting an alkaline pH and a significant (P = .02) increase in the numbers of RBCs and WBCs was observed in the diseased group compared to the control one. In the control group, white light cystoscopy showed a smooth pale pink glistening mucosa with two openings of the ureters and visible submucosal blood vessels. During chromocystoscopy, faint bluish discoloration of the mucosal surface with clearly visible submucosal blood vessels was detectable. These findings were correlated with the histopathological findings of the biopsies collected from the urinary bladder. In the diseased group, white-light cystoscopy showed clearly visible blood vessels, mildly hyperaemic mucosa in focal or diffuse forms and small vesicle formation. Chromocystoscopy revealed dark bluish oedematous and irregular mucosa either in a focal form or a diffuse form (marbled appearance) with deeply stained submucosal blood vessels. Histopathologically, the urothelium was hyperplastic with squamous metaplasia and the lamina propria was infiltrated with few leukocytes and congested blood vessels. Small bluish dots representing the absorbed methylene blue dye were seen in the inflamed areas against the lightly stained mucosa of the bladder. Severe diffuse necrotic cystitis was also seen with bacterial aggregations on the surface. Gram's staining revealed both gram positive bacilli and Gram positive coccobacilli. In conclusion, chromoendoscopy is a helpful tool for early diagnosis of cystitis in female donkeys and enables targeted biopsies, which improves the prognosis and outcome of therapy.

Novel approach to gastric mucosal defect repair using fresh amniotic membrane allograft in dogs (experimental study).

BACKGROUND: Gastric mucosal defect could result from several causative factors including the use of nonsteroidal anti-inflammatory drugs, Helicobacter pylori infection, gastrointestinal and spinal cord diseases, and neoplasia. This study was performed to achieve a novel simple, inexpensive, and effective surgical technique for the repair of gastric mucosal defect. METHODS: Six adult male mongrel dogs were divided into two groups (three dogs each). In the control positive group (C + ve), dogs were subjected to surgical induction of gastric mucosal defect and then treated using traditional medicinal treatment for such a condition. In the amniotic membrane (AM) group, dogs were subjected to the same operation and then fresh AM allograft was applied. Clinical, endoscopic, biochemical (serum protein and lipid and pepsin activity in gastric juice), histopathological, and immunohistochemistry evaluations were performed. RESULTS: Regarding endoscopic examination, there was no sign of inflammatory reaction around the grafted area in the AM group compared to the C + ve group. The leukocytic infiltration in the gastric ulcer was well detected in the control group and was less observed in the AM group. In the AM group, the concentrations of both protein and lipid profiles were nearly the same as those in serum samples taken preoperatively at zero time, which indicated that the AM grafting acted the same as gastric mucosa. The re-epithelization of the gastric ulcer in the C + ve group was not yet detected at 21 days, while in the AM group it was well observed covering most of the gastric ulcer. AM accelerated the re-epithelization of the gastric ulcer. The fibrous connective tissue and the precursor of collagen (COL IA1) were poorly detected in the gastric ulcer with AM application. CONCLUSION: Using fresh AM allograft for repairing gastric mucosal defect in dogs showed great impact as a novel method to achieve optimum reconstruction of the gastric mucosal architecture and restoration of pre-epithelial, epithelial, and post-epithelial normal gastric mucosal barriers.

Evaluation of subcutaneous infiltration of autologous platelet-rich plasma on skin-wound healing in dogs.

Platelet-rich plasma (PRP) is known to be rich in growth factors and cytokines, which are crucial to the healing process. This study investigate the effect of subcutaneous (S/C) infiltration of autologous PRP at the wound boundaries on wound epithelization and contraction. Five adult male mongrel dogs were used. Bilateral acute full thickness skin wounds (3 cm diameter) were created on the thorax symmetrically. Right side wounds were subcutaneously infiltrated with activated PRP at day 0 and then every week for three consecutive weeks. The left wound was left as control. Wound contraction and epithelization were clinically evaluated. Expression of collagen type I (COLI) A2, (COLIA2),histopathology and immunohistochemical (IHC) staining of COLI α1 (COLIA1) were performed on skin biopsies at first, second and third weeks. The catalase activity, malondialdehyde (MDA) concentration and matrix metalloproteinase (MMP) 9 (MMP-9) activity were assessed in wound fluid samples. All data were analysed statistically. The epithelization percent significantly increased in the PRP-treated wound at week 3. Collagen was well organized in the PRP-treated wounds compared with control wounds at week 3. The COLIA2 expression and intensity of COLIA1 significantly increased in PRP-treated wounds. MDA concentration was significantly decreased in PRP-treated wound at week 3. The catalase activity exhibited no difference between PRP treated and untreated wounds. The activity of MMP-9 reached its peak at the second week and was significantly high in the PRP-treated group. S/C infiltration of autologous PRP at the wound margins enhances the wound epithelization and reduces the scar tissue formation.

Enhancement of neural regeneration after spinal cord injury using muscle graft in experimental dogs.

AIMS: Spinal cord injuries (SCIs) can cause severe disability or death. The principal treatments for traumatic SCI include surgical stabilization and decompression. Using muscle as a scaffold is a new approach. The aim of this work is to evaluate the clinical efficacy of muscle graft as a scaffold for the growing axons organizing their growth, preventing gliosis in the damaged area and enhancing neural recovery in canine model of traumatic spinal cord injury. METHODS: 14 dogs were divided into group I (Control group) 4 control dogs subjected to Sham operation, group II (Trauma control group) 5 dogs subjected to dorsal laminectomy with excision of 1 cm segment of the spinal cord and group III (Muscle graft group) 5 dogs subjected to dorsal laminectomy then muscle graft was taken from the longissimus thoraces and inserted into the spinal cord gap. The animals of all groups were euthanatized after 8 weeks. Olby and modified Tarlov scores were used to clinically evaluate the therapeutic effects. Spinal cord specimens were subjected to histological, morphometric and statistical studies. RESULTS: Olby and modified Tarlov scores revealed significant clinical improvement in the muscle graft group. Histological sections showed overgrowth of axons on the muscle graft and the sections started to organize as central gray matter and peripheral white matter. CD44 & CD105 stains were positive for endogenous stem cells. CONCLUSIONS: This study proved the clinical efficacy of muscle grafting as a tool for induction of neuroregeneration after traumatic spinal cord injury.