Zdzisław Kiełbowicz

The carbon dioxide laser: an alternative surgery technique for the treatment of common cutaneous tumors in dogs

BackgroundTumors of the skin and subcutaneous tissue are the largest group of canine neoplasms. Total excision is still the most effective method for treatment of these skin tumors. For its universal properties the carbon dioxide (CO2) laser appears to be an excellent surgical instrument in veterinary surgery. Laser techniques are alternatives to traditional methods for the surgical management of tumors. The aim of this study was to compare various types of laser techniques in skin oncologic surgery: excision, ablation and mixed technique and to suggest which technique of CO2 laser procedure is the most useful in particular case of tumors in dogs.FindingsThe study was performed on 38 privately-owned dogs with total number of 40 skin tumors of different type removed by various CO2 laser operation techniques from 2010–2013. The treatment effect was based on the surgical wound evaluation, the relative time of healing and possible local recurrence of the tumor after 3 months post surgery. Local recurrence was observed in two cases. The study showed that in 30 cases time needed for complete resection of lesions was less than 10 minutes. Time of healing was longer than 12 days in 6 cases (42.8%) with tumor excision and in 14 cases (87.5%) where excision with ablation technique was performed.ConclusionsThe advantages of the CO2 laser surgery were better hemostasis, precision of working, non-contact dissection, less instruments at the site of operation and minimum traumatization of the surrounding tissues.

Antlerogenic stem cells: molecular features and potential in rabbit bone regeneration

ABSTRACT Aim: (i) To assess the expression profiles of stem cell-associated markers including Oct4, Sox2, Klf4, Nanog, C-myc, Stat3 and Cd9, (ii) analyze the nanotopography of the MIC-1 stem cells and (iii) evaluate the efficiency of live stem cell implants and stem cell culture derivatives on the regeneration of bone deficiencies in rabbit mandibles. Materials and methods: The expression profiles of stem cell-associated genes, including Oct4, Sox2, Klf4, Nanog, C-myc, Stat3 and CD9 were assessed using reverse transcription polymerase chain reaction and flow cytometry. Nanotopography of the antlerogenic MIC-1 cell lineage was analyzed using atomic force microscopy. The effect of MIC-1 stem cells, their homogenate and supernatant on the regeneration of bone deficiencies in rabbit mandibles was evaluated using histological analysis. The effect of MIC-1 stem cells and stem cell-based derivatives on the immune responses of the animals was assessed by analyses of acute phase protein levels (haptoglobin and fibrinogen). Results: We found that the MIC-1 cells isolated from the apical regions of growing antlers exhibited molecular features that were characteristics of pluripotent stem cells. Using atomic force microscopy, we determined the details of the cell surface morphologies with a particular emphasis on the patterns of formation of plasma extensions for interlinking adjacent cells. We also demonstrated that not only implanted stem cells but also cell homogenates and cell post-culture supernatants have potential in the regeneration of bone deficiencies in the rabbit mandible. Conclusions: Our findings indicate that the use of both antlerogenic stem cell implants and the preparations derived from the cells offer alternative approaches to those based on autologous stem cells in the biological stimulation of osteogenesis and in bone regeneration.

The Rabbit Brachial Plexus as a Model for Nerve Repair Surgery—Histomorphometric Analysis

One of the most devastating injuries to the upper limb is trauma caused by the avulsion. The anatomical structure of the rabbits brachial plexus is similar to the human brachial plexus. The aim of our study was to analyze the microanatomy and provide a detailed investigation of the rabbits brachial plexus. The purpose of our research project was to evaluate the possibility of utilizing rabbits plexus as a research model in studying brachial plexus injury. Studies included histomorphometric analysis of sampled ventral branches of spinal nerves C5, C6, C7, C8, and Th1, the cranial trunk, the medial part of the caudal trunk, the lateral part of the caudal trunk and peripheral nerve. Horizontal and vertical analysis was done considering following features: the axon diameter, fiber diameter and myelin sheath. The number of axons, nerve area, myelin fiber density and minimal diameter of myelin fiber, minimal axon diameter and myelin area was marked for each element. The changes between ventral branches of spinal nerves C5–Th1, trunks and peripheral nerve in which the myelin sheath, axon diameter and fiber diameter was assessed were statistically significant. It was found that the g‐ratio has close value in the brachial plexus as in the peripheral nerve. The peak of these parameters was found in nerve trunks, and then decreased coherently with the nerves travelling peripherally. Anat Rec, 298:444–454, 2015.

Dynamic tissue perfusion measurement in the reproductive organs of the female and male dogs

Abstract The aim of this study was to investigate the usefulness of new software Pixel Flux (PXFX) for clinical evaluation of tissue perfusion in the field of reproduction in dogs. The experiment was performed on six adult Beagle dogs. Different organs and tissues of the animals were examined with the MyLab25 Gold ultrasound system. Blood flow in the ovary, testicle, prostate, the ramification of the penile artery, and the network of blood vessels of the pampiniform plexus were examined with the use of colour coded Doppler technique, and obtained data was evaluated with the PXFX software. The more objective digital evaluation of data obtained with colour Doppler sonography through the application of dynamic tissue perfusion measurements provides new opportunities for diagnosis, as well as continuous monitoring of the function of the examined tissues and organs. The use of PXFX software is strongly indicated as a tool in small animal practice as an additional method for evaluation of tissue perfusion, especially in the cases when other methods like pulsed wave Doppler techniques are difficult to be performed

Prevalence of dorsal notch and variations in the foramen magnum shape in dogs of different breeds and morphotypes

Abstract The study evaluated shape of the foramen magnum (FM) in dog with regard to its constitutional type (small, medium and large breeds) and morphotype (brachycephalic, mesaticephalic and dolichocephalic dogs). We used multi-slice CT scanner to examine occipital area of 138 purebreds dogs. Two studied groups could be distinguished – living animals undergoing standard diagnostic CT procedure (n = 47) and macerated skulls of euthanized dogs (n = 91). Morphometric analysis was focused on presence of dorsal notch within foramen magnum. This anatomical feature was present in 33.3% of examined dogs. It was more often observed in small size dogs (70%) and those of brachycephalic morphotype (94.1%). For the first time dorsal notch was noted in breeds such as French Bulldog, Fox Terrier, Keeshond, Standard Schnauzer, English Bulldog, Cocker Spaniel, Boxer and Rottweiler. We distinguished four different shape of FM: oval, pentagonal, rhomboid and circular. Most common FM shape was oval (41 dogs; 45%) whereas circular type was least observed (5 dogs; 5.5%). The pentagonal shape was noticed in 27 specimens (29.7%) and the rhomboid shape in 18 dogs (19.8%). Authors conclude that dorsal notch is not pathology and should be considered a morphological variation within the normal anatomy. Results presented in this study should be taken into account during interpretation of the CT images of the craniocervical junction.

N-Arylaminomethylenebisphosphonates Bearing Fluorine Atoms: Synthesis and Antiosteoporotic Activity

GRAPHICAL ABSTRACT Abstract A series of N-phenyl and N-pyridyl-aminomethylenebisphosphonates substituted in their aromatic rings with fluorine atoms or trifluoromethyl groups have been synthesized by a three component approach. They were screened for potential antiosteoporetic activity using mouse macrophage-like J774E cells. Most of the compounds appeared to be moderate inhibitors of macrophage cells proliferation compared to known the antiosteoporetic drug, Incandronate. Their potency was compared with their structural analogues, hydroxymethylene-bisphosphonate 2 and ethylidenebisphosphonates 3, which appeared to be equipotent. Although the aminomethylenebisphosphonates 1 show low stability in aqueous solutions, the most stable, i.e., N-(3-trifluoromethylphenyl)-aminomethylenebisphosphonic acid, was chosen for in vivo testing on sheep with induced osteoporosis. This compound had marginal influence on the bone structure recovery.

Effect of mucoprotective plant-derived therapies on damage to colonic mucosa caused by carprofen and robenacoxib administered to healthy dogs for 21 days

Background: Non-steroidal anti-inflammatory drugs (NSAIDs) may cause gastrointestinal damage in dogs. Hypothesis/objectives: To determine the extent to which lansoprazole, liquorice extract, and a herbal solution exhibit protective effects on colonic mucosa when administered to dogs concurrently with the NSAIDs carprofen or robenacoxib. Animals and methods: Thirty-five healthy beagle dogs (15 male and 20 female) aged 13–14 weeks and weighing 4.3–5.5 kg at the beginning of the experiment were included. Endoscopy and biopsy of the caudal gastrointestinal tract were performed pretreatment and on the last day of a 21-day treatment period with (1) oral carprofen; (2) carprofen and the proton-pump inhibitor lansoprazole; (3) carprofen, liquorice extract, and a herbal solution that contained extracts of thyme, icelandic lichen, hyssop, and saponariae root; (4) robenacoxib; (5) robenacoxib and lansoprazole; (6) robenacoxib, liquorice extract, and herbal solution; or (7) an empty gelatin capsule. Statistical analyses were performed with the Kruskal–Wallis, Cochrans Q, and chi-squared test with p < 0.05 considered significant. Results: Both carprofen and robenacoxib tested damaged the colonic mucosa with most severe microscopic lesions following administration of robenacoxib with lansoprazole. The risk of histopathological lesions in the colon increased most rapidly in robenacoxib with lansoprazole (absolute risk increase −0.85) similar to robenacoxib only (−0.75), whereas the best result was recorded following the plant remedies together with carprofen (−0.15) and the plant remedies together with robenacoxib (−0.2). Conclusions and clinical importance: Concurrent administration of liquorice extract and an herbal solution with robenacoxib was associated with decreased severity of the NSAID-induced mucosal lesions.

Hormone Receptor Expression Analyses in Neoplastic and Non-Neoplastic Canine Mammary Tissue by a Bead Based Multiplex Branched DNA Assay: A Gene Expression Study in Fresh Frozen and Formalin-Fixed, Paraffin-Embedded Samples.

Immunohistochemistry (IHC) is currently considered the method of choice for steroid hormone receptor status evaluation in human breast cancer and, therefore, it is commonly utilized for assessing canine mammary tumors. In case of low hormone receptor expression, IHC is limited and thus is complemented by molecular analyses. In the present study, a multiplex bDNA assay was evaluated as a method for hormone receptor gene expression detection in canine mammary tissues. Estrogen receptor (ESR1), progesterone receptor (PGR), prolactin receptor (PRLR) and growth hormone receptor (GHR) gene expressions were evaluated in neoplastic and non-neoplastic canine mammary tissues. A set of 119 fresh frozen and 180 formalin-fixed, paraffin-embedded (FFPE) was comparatively analyzed and used for assay evaluation. Furthermore, a possible association between the hormone receptor expression in different histological subtypes of canine malignant mammary tumors and the castration status, breed and invasive growth of the tumor were analyzed. The multiplex bDNA assay proved to be more sensitive for fresh frozen specimens. Hormone receptor expression found was significantly decreased in malignant mammary tumors in comparison to non-neoplastic tissue and benign mammary tumors. Among the histological subtypes the lowest gene expression levels of ESR1, PGR and PRLR were found in solid, anaplastic and ductal carcinomas. In summary, the evaluation showed that the measurement of hormone receptors with the multiplex bDNA assay represents a practicable method for obtaining detailed quantitative information about gene expression in canine mammary tissue for future studies. Still, comparison with IHC or quantitative real-time PCR is needed for further validation of the present method.

Feasibility and accuracy of new insertion technique of S1 transpedicular screw. Computed tomography-based morphometric analysis

OBJECTIVE To assess feasibility and accuracy of a new insertion technique of S1 transpedicular screw. SUMMARY OF BACKGROUND DATA Transpedicular stabilization in the first sacral vertebra (S1) is a technically demanding surgical procedure with inherent risk of loosening of the implant. A modification of the technique was recently proposed, along with the analytical verification which was performed based on the available literature. In the study, we performed radiological assessment of screws inserted into the S1 using the classical and modified techniques. METHODS The analysis was performed in two parts. The first part was performed on eight cadaver specimens after implantation of the screws. In the second part, we used computed tomography images of patients with degenerative disk disease with a superimposed representation of screws. The thickness of the posterior cortex adherent to the screws, screw trajectory and their position with regard to the spinal canal was measured. The area of posterior cortex in contact with the screws was also calculated. RESULTS The contact length and area was found to be two times greater for screws introduced with the modified technique. The convergence angle was comparable between the techniques, despite the shift of entry point. There was no canal breach, although with the modified technique the screws passed closer to the spinal canal. CONCLUSIONS The modified technique is considered safe. In this technique, the screws pass through a thicker portion of the posterior cortex compared to the classical technique that aims at improving the stability of the fixation.

Improvement of large animal model for studying osteoporosis

Abstract The aim of the study was to determine the impact of steroidal medications on the structure and mechanical properties of supporting tissues of sheep under experimentally-induced osteoporosis. A total of 21 sheep were used, divided into three groups: a negative control (KN) (n = 3), a positive control (KP) (n = 3) with ovariectomy, and a steroidal group (KS) (n = 15) with ovariectomy and glucocorticosteroids. All animals were kept on a low protein and mineral diet and had limited physical activity and access to sunlight. Quantitative computed tomography was the examination method. The declines in the examined parameter values in the KS group were more than three times higher than in the KN group. The study suggests that a glucocorticosteroidal therapy accelerates and intensifies processes taking place in the course of osteoporosis. The combination of glucocorticosteroids with ovariectomy, a restrictive diet, limited physical activity, and no access to sunlight leads to a decrease in radiological bone density.