Additional file 1: of Rostral cranial fossa as a site for cerebrospinal fluid drainage – volumetric studies in dog breeds of different size and morphotype

Individuals value of AP, ZyZy, volRCF, volCC and volRCF/volCC ratio (RCF/CC). (XLS 20 kb

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

<p><b><i>Aim</i>:</b> (i) To assess the expression profiles of stem cell-associated markers including <i>Oct4, Sox2, Klf4, Nanog, C-myc, Stat3</i> and <i>Cd9</i>, (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. <b><i>Materials and methods</i>:</b> The expression profiles of stem cell-associated genes, including <i>Oct4, Sox2, Klf4, Nanog, C-myc, Stat3</i> and <i>CD9</i> 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). <b><i>Results</i>:</b> 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. <b><i>Conclusions</i>:</b> 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.</p

<i>N</i>-Arylaminomethylenebisphosphonates Bearing Fluorine Atoms: Synthesis and Antiosteoporotic Activity

<div><p></p><p>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.</p></div

Laparoscopic intraoperative view of the anastomosis of the ureter with the bladder wall and the obtained postoperative result.

A: Placing a suture between the ureter and bladder wall. Ectopic ureter (EU) after transposition into a new opening in the bladder (B). The blue arrow indicates the needle used during suturing. B—View of the performed anastomosis: blue arrows indicate the suture site. C: View of the postoperative wounds after the working trocars (blue arrows) and the optic trocar (red arrow); the yellow arrow indicates the cranial part of the patient. D—View of the transpositioned ureter (blue arrow) two months after surgery during control cystoscopy in a female dog with persistent urinary incontinence.</p

Fig 1 -

Cystoscopic assessment, patient positioning for surgery and stages of ectopic ureter preparation for laparoscopic ureteroneocystostomy: A: Cystoscopic view of an ectopic ureter (lower left corner) opening into the distal urethra (UR). B—Patient positioning for the laparoscopic ureteroneocystostomy in lateral recumbency with the affected (ectopic) side up;—the blue arrow points to the cranial part of the operated animal. C: Trocar placement during laparoscopic ureteroneocystostomy. Working trocars, blue arrows; optic trocar, red arrow. D: Laparoscopic intraoperative view of the ectopic ureter (EU) before dissection from the surrounding tissues. B, urinary bladder; UT, uterus; R, rectum. E, F: Laparoscopic operative view of the subsequent stages of ectopic ureter (EU) preparation from the surrounding tissues. BL: Bladder lateral ligament. During preparation, the grasp has to be of the tissues surrounding the ureter, not the ureter itself, to avoid damage. The blue arrow shows the correct way to hold the ureter when preparing.</p

Fig 2 -

Laparoscopic intraoperative view of different stages of the ectopic ureter transposition: A—Ligation of the ectopic ureter (EU) and sliding knot (blue arrow); B, the urinary bladder. B—Red arrow indicates vessel coagulation on the surface of the ectopic ureter (EU) at the level of the planned transverse ureter incision. The blue arrow indicates the site of the previous ligation. B—urinary bladder. C: Transverse cut of the ureter (red arrow) near the previously placed ligation (blue arrow). EU: ectopic ureter. B: urinary bladder. D: Ureter is cut parallel to its long axis before transposition into the bladder lumen (red arrow); EU: ectopic ureter; B: urinary bladder. E—Placing the dissected and cut-off ectopic ureter (EU) to determine a spot for transposition to the bladder (B), which reduces the risk of excessive tension at the place of subsequent anastomosis. The red arrow indicates the laparoscopic hook and designated ureteral transposition site. F: Orifice in the bladder (B) wall of the transposed ureter (B). The diameter of the orifice was approximately equal to the outer diameter of the ureter. The red arrow indicates the bladder lumen after mucosal incision.</p

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

<div><p>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.</p></div

List of genes and their respective Accession Number and Probe Set Region used in the QuantiGene 2.0 Plex Assay.

<p>List of genes and their respective Accession Number and Probe Set Region used in the QuantiGene 2.0 Plex Assay.</p

Normalized gene expression of PRLR in fresh frozen samples.

<p>Asterisks indicate statistical significance (* p<0.05; ** p<0.01; *** p<0.001; **** p<0.0001). The box encloses cases within the 25^th to the 75^th percentiles. The horizontal line within the box represents the median and the upper and lower bars are the largest and lowest observed values. Samples with a value higher than four standard deviations above the mean are not shown in the graph.</p

Median of normalized gene expression of ESR1 and PGR in intact (fresh frozen n = 57; FFPE n = 83) and neutered (fresh frozen n = 11; FFPE n = 8) females with malignant mammary tumors.

<p>Median of normalized gene expression of ESR1 and PGR in intact (fresh frozen n = 57; FFPE n = 83) and neutered (fresh frozen n = 11; FFPE n = 8) females with malignant mammary tumors.</p