Laparoscopic ureteroneocystostomy in the treatment of urinary incontinence due to ectopy of the ureters in female dogs: A pilot study.

Ureteral ectopia is rare and requires surgical treatment after a thorough diagnostic workup. Open surgical techniques for repositioning ectopic ureters have been known for many years and are well described in the literature. However, to the best of our knowledge, no laparoscopic method of correcting this pathology has been described, which, in our opinion, would benefit the animal in terms of the healing process and overall clinical outcomes. This study aimed to evaluate the possibility of laparoscopic treatment of ureteral ectopia, which causes urinary incontinence in dogs. All of the operated ten dogs presented in this study were client-owned females with symptoms of urinary incontinence due to a unilateral intramural ectopic ureter. A three-trocar laparoscopic technique was used to perform the ureteroneocystostomy of the ectopic ureter. In this article, clinicopathological data, imaging features, procedural findings, complications, and short- and long-term outcomes are presented. The procedure was feasible in all cases. No major postoperative complications were observed. Among the minor complications, slight hematuria was observed in three dogs, which resolved spontaneously. In the period of at least one year after surgery, no negative impact of the procedure was observed. Seven of the ten operated dogs regained urinary continence. The remaining three dogs required additional surgery (urethral bulking) because of a lack of improvement after adjuvant pharmacological treatment. Overall, good-to-excellent long-term outcomes can be achieved; however, dogs that remain incontinent after laparoscopic ureteroneocystostomy may require additional treatment.

Preliminary study on fluid bolus administration for prevention of spinal hypotension in dogs undergoing elective cesarean section.

Introduction: This study aimed to investigate the effect of fluid bolus administration during epidural anesthesia (coload) in female dogs scheduled for elective cesarean section (CS). Hypotension is one of the most common complications of epidural (EA) and spinal (SA) analgesia, and in the case of cesarean section, it may pose a significant risk for placental perfusion and subsequent fetal vitality and puppy survival. Methods: Pregnant bitches scheduled for elective CS underwent EA with (treatment group) or without (control group) intravenous fluid bolus administration. The following parameters were measured and compared between both groups: HR, RR, etCO2, SpO2, systolic, diastolic and mean arterial blood pressure were measured at three time points (T1: before surgery, T2: after the last puppy removal, and T3: end of surgery) in dams; vitality (Apgar score at 0, 5, and 20 min) and umbilical cord blood parameters (pH, pCO2, HCO3, base excess, lactate and glucose) in newborns. Results: The results indicated that crystalloid coloading increased maternal systolic, diastolic, and mean blood pressure (treatment vs. control group, 101.46 ± 9.18, 48.01 ± 13.47, and 67.07 ± 13.15 mmHg vs. 80.68 ± 7.29, 36.52 ± 8.75, and 180 52.30 ± 7.77, p < 0.05) with significantly fewer episodes of hypotension. Additionally, puppies in the treatment group received higher scores in the 5-min (7.91 ± 1.67 vs. 6.74 ± 2.20) and 20-min (9.38 ± 0.87 vs. 8.39 ± 2.50) assessments without the favorable effect on umbilical blood gas parameters. Discussion: Based on the obtained results, it can be stated that crystalloid coload offers an effective option in cases of hypotension during cesarean section, with clear benefits for both mothers and newborns.

A Novel Surgical Hybrid Approach to Neoplastic Lesions in the Distal Part of the Urethra: A Pilot Series of Clinical Cases.

All the surgical approaches described to date for the removal of distal urethral tumors have some technical difficulties that make these tumors difficult to treat. The article presents for the first time the treatment results of three female dogs, diagnosed with transitional cell carcinomas of the distal urethra, operated with a newly developed hybrid surgical method-a combination of laparoscopy and open surgery. This technique uses vulvovaginectomy, combined with resection of the distal urethra and prepubic urethrostomy. All of the procedures were possible to perform, without the need to carry out a laparotomy conversion. Histopathology revealed transitional cell carcinoma in all cases, with a margin of healthy tissues maintained in two out of three cases, which meant reoperation of the urethrostomy site in the remaining one case. The mean survival time was nine months. Among minor complications, recurrent cystitis was found. After the first surgery, all dogs retained full control over urination immediately after recovery from anesthesia. In one case that required reoperation, complete urinary incontinence occurred after the second procedure. The present findings suggest that hybrid surgery can be used to treat distal urethral tumors. Qualification for surgery must be limited to bitches with tumors of the distal urethra and without metastases, without the possibility of using other surgical methods, and with the owner's full acceptance of the risk of complications.

Comparison of 2 anesthetic protocols and surgical timing during cesarean section on neonatal vitality and umbilical cord blood parameters.

BACKGROUND: The objective of this study was to evaluate the relationship between the mode of anesthesia, the time form the induction to the extraction of a puppy and the immediate postnatal vitality and umbilical cord blood gases parameters in cesarean section derived-puppies. Two different anesthetic protocols were used: inhalation using isoflurane (ISO) and combined-inhalation and epidural (EPI) with propofol being the induction agent. RESULTS: Significant differences were found in ISO group in pH values, pCO2 levels and Apgar scores between puppies at different extraction times (< 30 vs. ≥ 30 min). In ISO group puppies extracted later were more acidic (7.16 vs. 7.22), had higher levels of pCO2 (69 vs. 57 mmHg) and lower Apgar scores at birth (1.2 vs. 2.5). On the contrary, in EPI group no differences were observed between the delivery time, umbilical blood gas parameters and puppies' vitality. Furthermore, the dams from the EPI group required lower concentrations of isoflurane (MAC 1.11 ± 0.19 vs.1.37 ± 0.16, p < 0.001). CONCLUSIONS: Multiple pregnancies frequent in dogs lead to significant differences in extraction times between the first and the last puppy during cesarean section. Obtained results showed that the mode of anesthesia and the surgical time would influence the neonatal outcome during cesarean section in dogs. The higher concentration of isoflurane with the longer time of exposure had a negative effect on the initial newborn vitality as well as the umbilical cord blood gas parameters. Therefore, when performing CS in giant dog breeds or expecting many puppies in the litter, it is worth considering epidural component that allow for lower concentrations of inhalant agents, which may contribute to a better clinical condition of newborns.

Osteogenic Potential of Sheep Mesenchymal Stem Cells Preconditioned with BMP-2 and FGF-2 and Seeded on an nHAP-Coated PCL/HAP/ß-TCP Scaffold.

Mesenchymal stem cells (MSCs) attract interest in regenerative medicine for their potential application in bone regeneration. However, direct transplantation of cells into damaged tissue is not efficient enough to regenerate large bone defects. This problem could be solved with a biocompatible scaffold. Consequently, bone tissue engineering constructs based on biomaterial scaffolds, MSCs, and osteogenic cytokines are promising tools for bone regeneration. The aim of this study was to evaluate the effect of FGF-2 and BMP-2 on the osteogenic potential of ovine bone marrow-derived MSCs seeded onto an nHAP-coated PCL/HAP/ß-TCP scaffold in vitro and its in vivo biocompatibility in a sheep model. In vitro analysis revealed that cells preconditioned with FGF-2 and BMP-2 showed a better capacity to adhere and proliferate on the scaffold than untreated cells. BM-MSCs cultured in an osteogenic medium supplemented with FGF-2 and BMP-2 had the highest osteogenic differentiation potential, as assessed based on Alizarin Red S staining and ALP activity. qRT-PCR analysis showed increased expression of osteogenic marker genes in FGF-2- and BMP-2-treated BM-MSCs. Our pilot in vivo research showed that the implantation of an nHAP-coated PCL/HAP/ß-TCP scaffold with BM-MSCs preconditioned with FGF-2 and BMP-2 did not have an adverse effect in the sheep mandibular region and induced bone regeneration. The biocompatibility of the implanted scaffold-BM-MSC construct with sheep tissues was confirmed by the expression of early (collagen type I) and late (osteocalcin) osteogenic proteins and a lack of an elevated level of proinflammatory cytokines. These findings suggest that FGF-2 and BMP-2 enhance the osteogenic differentiation potential of MSCs grown on a scaffold, and that such a tissue engineering construct may be used to regenerate large bone defects.

Reduction of greenhouse gases emission through the use of tiletamine and zolazepam.

Isoflurane is an anaesthetic gas widely used in both human and veterinary medicine. All currently used volatile anaesthetics are ozone-depleting halogenated compounds. The use of total intravenous anaesthesia (TIVA) allows to induce the effect of general anaesthesia by administering drugs only intravenously without the use of anaesthetic gases. This allows you to create a protocol that is safe not only for the patient, but also for doctors and the environment. However, so far, no anaesthetic protocol based on induction of anaesthesia with tiletamine-zolazepam without the need to maintain anaesthesia with anaesthetic gas has been developed. Our study showed that the use of this combination of drugs for induction does not require the use of additional isoflurane to maintain anaesthesia. Thanks to Dixon's up-and-down method we proved that with the induction of anaesthesia with tiletamine-zolazepam at a dose of 5 mg/kg the use of isoflurane is not needed to maintain anaesthesia in minimally invasive surgical procedures. Until now, this dose has been recommended by the producer for more diagnostic than surgical procedures or for induction of general anaesthesia. The maintenance was required with anaesthetic gas or administration of another dose of the tiletamine-zolazepam. The results obtained in this study will allow for a significant reduction in the consumption of isoflurane, a gas co-responsible for the deepening of the greenhouse effect, having a negative impact on patients and surgeons. These results are certainly the first step to achieving a well-balanced and safe TIVA-based anaesthetic protocol using tiletamine-zolazepam, the obvious goal of which will be to maximize both the safety of the patient, people involved in surgical procedures, and the environment itself. Being aware of the problem of the greenhouse effect, we are committed to reducing the consumption of anaesthetic gases by replacing them with infusion agents.

Is an epidural component during general anaesthesia for caesarean section beneficial for neonatal puppies' health and vitality?

Regional centro-axial block is a recommended technique for uncomplicated caesarean section in human medicine. Since the application of regional anaesthesia as the only technique in veterinary medicine is impractical, the objective of the study was to assess and compare the epidural component of caesarean section (CS) on maternal and fetal outcomes. Bitches (n = 36) undergoing elective CS were enrolled in this study. Females were randomly assigned into two groups: Gr I (Isoflurane, n = 20) and Gr IE (Isoflurane plus Epidural, n = 16). Anaesthesia was induced with propofol, and maintained with isoflurane in oxygen. In the IE group, epidural anaesthesia was also performed using lidocaine. The maternal intraoperative parameters were compared at three time points: T1 - just before the skin incision, T2 - after the last puppy removal, and T3 - at the end of surgery. At least 100 mcl of mixed umbilical cord blood was collected for gas analysis. The modified Apgar scoring system (AS) was used to objectively score newborn health and vitality immediately after birth (0 min), 5 and 20 min after birth. Systolic, diastolic, and mean blood pressure were lower in the IE group at T1, T2, and T3 compared to I group (p < 0.05). In the combined anaesthesia group blood pressure remained stable but low at all time points. Throughout surgery, the IE group required a lower concentration of isoflurane (p < 0.05). The median values of the umbilical blood gas results were found to be similar (p > 0.05) in both investigated groups. The initial results of the Apgar score were comparable in the I and IE groups (p > 0.05). However, subsequent AS measurements revealed significant differences between both groups. Puppies from the IE group received better AS scores at 5 and 20 min compared to the I group (median AS 8 and 9 vs. 5 and 8, respectively). The results obtained demonstrated that epidural anaesthesia administration reduced the requirement for isoflurane in dams undergoing caesarean section and despite episodes of maternal hypotension did not affect the results of neonatal umbilical blood gas. Furthermore, newborns from the epidural anaesthesia group improved more quickly postnatally, developing a satisfactory condition in a shorter time.

Indirectly assessing changes in corneal properties with OCT speckle after crosslinking in porcine eyes.

The aim of this study was to assess the effect of the standard crosslinking (CXL) procedure on corneal properties and subsequent changes in collagen bonds formation using optical coherence tomography (OCT) corneal speckle statistics and vibrational spectroscopy. Porcine eyes with intact corneal epithelium were randomly selected to one of the four study groups: (1) untreated eyes moistened with phosphate-buffered saline (PBS); (2) eyes after the epithelial debridement and riboflavin application; (3) eyes after CXL procedure according to the Dresden protocol; and (4) eyes after corneal epithelial debridement, regularly moistened with PBS. Before and after this selection, each eyeball was subjected to the constant intraocular pressure of 20 mmHg. Then, ocular biometry was performed and the central cornea was imaged using spectral-domain OCT. Following this, a nonparametric approach to speckle modeling (the Contrast Ratio (CR)) was utilized within the region of interest for each B-scan covering the central corneal stroma. To verify whether the CXL performed ex-vivo results in formation of new bonds in the cornea, Fourier Transform Infrared Attenuated Total Reflectance (ATR-FTIR) spectra of dried corneas, dissected from examined eyeballs, were collected and analyzed. Corneal epithelium removal alone or with the riboflavin application leads to a statistically significant decrease in the CR median value (the Wilcoxon signed-rank test, p < 0.05). However, the most pronounced change in CR median value, which decreases with the increased number of scatterers, was shown after the complete CXL procedure including riboflavin soaking and UVA irradiation (the Wilcoxon signed-rank test, p = 0.004). Analysis of ATR-FTIR spectra revealed influence of UVA irradiation on collagen matrix. The study has shown the increased dehydration accompanied by almost no alteration of collagen native triple-helical structure. Significant changes have been observed for bands related to collagen crosslinks. Specifically, the predominant changes occurred in the sugar region from 1150 to 975 cm-1, as well as in the absorbance of carbonyl groups. Furthermore, the ratio of two Amide I components at approximately 1660 cm-1 and 1675 cm-1 decreased after UVA irradiation. Together, these results provided the evidence for the creation of new corneal crosslinks. In conclusion, this study clearly indicates that the UVA exposure causes the substantial difference in optical scattering occurring in corneal stroma as a result of the induced biochemical changes at the molecular level in this tissue assessed with ATR-FTIR. The proposed speckle-based methodology brings a new insight into the development of OCT technology useful in an indirect assessment of some collagen changes.

Clinical Usefulness of the Anaesthetic Protocol Using Low Doses of Tiletamine - Zolazepam Combination in Continuous Rate Infusion as a Partial Intravenous Anaesthesia in Bitches Undergoing Laparoscopic Surgery.

The objective of this study was to evaluate the clinical usefulness of the combination of tiletamine-zolazepam used in low doses as a continuous rate infusion in a partial intravenous anesthesia protocol. Fifteen clinically healthy, different breed bitches weighing 25.08 ± 10.39 kg was used in this study. After a food fast for at least 12 hours and water fast for 4 hours, the animals were premedicated with dexmedetomidine. After 15 minutes, the bolus of tiletamine-zolazepam combination was given as an. Induction of general anesthesia, immediately followed by continuous intravenous infusion. The following parameters were measured immediately after the induction of general anesthesia and lasted until the end of the surgery: electrocardiography, heart rate, systolic arterial blood pressure, diastolic arterial blood pressure, mean arterial blood pressure, body temperature respiratory rate end tidal of CO2. During the recovery period, pain level was evaluated as well as sedation assessment. Time for successful intubation after administration of the tiletamine-zolazepam combination was within 3 minutes. Heart rate was within reference values. Systolic, diastolic, and mean arterial blood pressure were also within the reference values. Internal body temperature showed a downward trend for a whole procedure time. During recovery, only 1 patient showed symptoms of pain and signs of dissociation. In summary, the partial intravenous protocol with the use of tiletamine-zolazepam combination and low anesthetic gases concentration is clinically useful because of ensuring the correct level of anesthesia and stability of intraoperative parameters as well as a good recovery period.

Novel surgical approach to neoplastic lesions in the distal part of the urethra: A pilot cadaver study comparing open and hybrid techniques.

Tumours of the distal urethra in female dogs are often difficult to treat, and the surgical methods described thus far have technical limitations. This study aimed to present a novel approach to the surgical treatment of distal urethral tumours. This study used dog cadavers to evaluate the technical feasibility of surgically removing neoplastic lesions in the distal urethra and compared surgical outcomes of open surgery with those of hybrid surgery (combination of laparoscopy and open surgery). Open intact, open spayed, hybrid intact, and hybrid spayed dog cadaver groups underwent surgery (n = 6 per group). The novel surgical method was based on vulvovaginectomy (ovariohysterectomy in intact dogs), resection of the distal part of the urethra, and pre-pubic urethrostomy. Outcomes of interest included technical feasibility of each procedure, using both surgical techniques, wound length, time required to complete the procedure, and the incidence of intraoperative ureter and rectum injuries. Surgical technique and reproductive status affected operating time. Technique choice affected wound length; the surgical wound was longer in the open group than in the hybrid group. Macroscopic evaluation of the rectum did not reveal any damage to the wall. There was no evidence of ureter leakage or obstruction in any case. The present findings suggest that both open and hybrid surgery can be used to treat distal urethral tumours.

Parameters of Hemostasis in Sheep Implanted with Composite Scaffold Settled by Stimulated Mesenchymal Stem Cells-Evaluation of the Animal Model.

Implantation of composite scaffolds could be potentially associated with the risk of hemostatic disturbances in a recipient. However, there is a lack of information on possible alterations in clotting mechanisms resulting from such a procedure. The aim of the present work was to investigate changes in hemostatic parameters in sheep implanted with a scaffold composed of poly(ε-caprolactone) and hydroxyapatite and tricalcium phosphate (9:4.5:4.5), settled previously with mesenchymal stem cells stimulated by fibroblast growth factor-2 and bone morphogenetic protein-2. Nine Merino sheep were examined for 7 days, and measurements of clotting times (PT, aPTT), activities of antithrombin, protein C and clotting factors II-XII, and concentrations of fibrinogen and D-dimer were carried out before and 1 h, 24 h, 3 days and 7 days after scaffold implantation. The introduction of scaffold initially resulted in a slowdown of the clotting processes (most evident 24 h after surgery); PT and aPTT increased to 14.8 s and 33.9 s, respectively. From the third day onwards, most of these alterations began to return to normal values. The concentration of fibrinogen rose throughout the observation period (up to 8.4 g/L), mirroring the ongoing inflammatory reaction. However, no signals of significant disturbances in hemostatic processes were detected in the sheep tested.

Laparoscopic-assisted percutaneous herniorrhaphy as an alternative to open surgery technique in farm swines.

BACKGROUND: Despite numerous experimental studies presenting laparoscopic treatment of inguinal hernia in a pig model so far no described technique has been used in clinical patients of this species. Minimal invasiveness and the simplicity of closure of the inguinal canal using the Percutaneous Internal Ring Suturing (PIRS) technique makes it the world's first technique for laparoscopic treatment of inguinal hernia in pigs as clinical patients. AIM: This study aims to assess the applicability and effectiveness of the laparoscopic PIRS technique in the treatment of inguinal hernia in pigs as clinical patients and to compare the PIRS technique with the open surgery technique, which is currently being used. METHODS: The study was conducted on 22 non-castrated male pigs with inguinal hernia (clinical patients), divided into two equal groups: PIRS and open surgery (OS). In the PIRS group, the inner inguinal ring was closed with an optical trocar inserted at the umbilicus level and an injection needle with a suture material inserted percutaneously over the inguinal canal. The suture material was threaded through the inner inguinal ring and then tied, leaving the knot under the skin. As a result to this the inguinal canal was closed. In the OS group the procedure was performed with open access above the inguinal canal where, after dissection of the vaginal processus and reducing the contents of the hernia to the abdominal cavity, it was ligated as close to the inguinal canal as possible, and the wound was then closed in layers. RESULTS: All operated pigs returned to full fitness immediately after recovery from anesthesia. There was one case of hernia recurrence in the PIRS group. In the OS group all the operated pigs had a temporary swelling of the postoperative wound and the scrotum on the side of the operated inguinal hernia, which was not found in the PIRS group. CONCLUSIONS: The effectiveness of the PIRS technique is comparable to that of open surgery. Considering the simplicity of the PIRS procedure and its minimal invasiveness, this technique may be used as an alternative to the open technique in the treatment of inguinal hernias in pigs not subjected to surgical castration.

New therapeutic strategy: Personalization of pancreatic cancer treatment-irreversible electroporation (IRE), electrochemotherapy (ECT) and calcium electroporation (CaEP) - A pilot preclinical study.

In this study, irreversible electroporation (IRE), electrochemotherapy (ECT), and calcium electroporation (CaEP) techniques were investigated as new strategies for human pancreatic cancer. Qualification of the patients, best "therapeutic moment" for each patient, safety, and complications after procedures were examined. In this pilot study were included 13 patients in this study, which were operated on in different pancreatic cancer stages. Patients underwent IRE or ECT with intravenous admission of cisplatin or electroporation with calcium intratumoral administration. The IRE procedure was safe for the patients. Medium overall survival for IRE, IRE + CTH, and IRE + CaCl2 was respectively: 16, 29.5, and 19 months comparing to 10 months in control chemotherapy (CTH) group. Thus, IRE, ECT, and CaEP can be effective strategies for pancreatic cancer treatment.

Corneal Optical Coherence Tomography Speckle in Crosslinked and Untreated Rabbit Eyes in Response to Elevated Intraocular Pressure.

Purpose: To ascertain the influence of intraocular pressure (IOP) on corneal optical coherence tomography (OCT) speckle in untreated and ultraviolet A-riboflavin induced corneal collagen crosslinked rabbit eyes. Methods: Left corneas of eight rabbits were de-epithelialized and crosslinked by applying riboflavin and 30-minute ultraviolet A light exposure. After enucleation (6 months after treatment), each eyeball (treated and untreated) was mounted in a measurement setup, in which IOP was increased from 15 to 45 mm Hg in steps of 5 mm Hg. At each IOP value, single B-scans of the central cornea were acquired three times with the spectral-domain OCT Copernicus-HR. Then, three regions of interest, including the anterior, posterior, and entire corneal stroma, were automatically extracted. Five different probability distributions were used as a model for the corneal speckle and the one with the best goodness of fit was chosen for further analysis. Results: The generalized gamma distribution achieved the best goodness of fit and its scale (a) and shape (v) parameters statistically significantly changed with increasing IOP in the three regions of analysis (two-way repeated measures analysis of variance, all P < 0.05). The statistically significant difference between untreated and crosslinked eyes was observed for the shape parameters of the posterior and entire corneal stroma. Conclusions: Corneal OCT speckle is influenced by IOP and shows to be significantly different in untreated and crosslinked eyes. Corneal OCT speckle analysis has the potential to be indirectly used for assessing changes in corneal stroma in ex vivo and in vivo studies. Translational Relevance: Investigation of corneal OCT speckle statistics can offer additional diagnostic biomarkers related to changes in the corneal stroma after ocular surgeries.

Umbilical Cord Blood Gas Parameters and Apgar Scoring in Assessment of New-Born Dogs Delivered by Cesarean Section.

The article presents the results of the clinical evaluation (Apgar scores, AS) and umbilical cord blood gas analysis (UCBGA) obtained from clamped umbilical cords of newborn pups delivered by the elective Caesarean section. The study was planned as a controlled clinical study, the newborns were allocated into one of the groups, I-critical neonates (severe distress, AS ≤ 3), II-weak neonates (moderate distress, AS 4-6) and III-healthy neonates (no distress, AS ≥ 7). The following parameters were evaluated: pH (pH units), carbon dioxide partial pressure (pCO2; mmHg), oxygen partial pressure (pO2; mmHg), actual bicarbonate (cHCO3-; mmol/L), total carbon dioxide (cTCO2; mmol/L), base excess of extracellular fluid (BE(ecf); mmol/L), base excess of blood (BE(b); mmol/L), oxygen saturation (csO2; %), lactate (Lac; mg/dL), hematocrit (Hct; %PCV), hemoglobin (cHgb; g/dL), glucose (Glu; mg/dL), ions (Na, K, Ca, Cl). The majority of puppies had low AS at birth (AS 4-6 in 38.1% and AS ≤ 3 in 57.1% of the neonates), but most of them (85.7%) improved by the 20th min. reaching AS of 7 and more. Moreover, puppies with lower AS (≤ 3) were at higher risk of death within the first 24h (20.8% did not survive). The positive correlation was found between Apgar score measured at 0 min and pH (r = 0.46, p = 0.01), and between Apgar score (at 0 min) and base excess in whole blood measured [BE(b)] r = 0.36, p = 0.03). Whereas, a negative correlation was detected between Apgar score at 0 and 5th min and glycemia (r = -0.42, p = 0.05, r = -0.34, p = 0.02 respectively. Overall, the puppies with higher glucose levels had lower Apgar scores and were at higher risk of death. Furthermore, in our study, the newborn puppies had mild acidemia with elevated pCO2 levels and the HCO3 at the lower range of normal limits, suggesting the mixed component in the acidemic state. Adaptation to extra-uterine life is crucial and any practical improvement in neonatal diagnostics and care would be beneficial for newborn puppy survival.

The effect of changes in cardiovascular activity on corneal biomechanics and pulsation in rabbits.

The aim was to assess the relationships between cardiovascular activity, corneal pulse characteristics, and corneal biomechanics in rabbits. Seventeen rabbits were randomly assigned to one of two anesthetic regimens to induce differences in arterial blood pressure and heart rate. Experimental protocol included measuring blood flow parameters in the ophthalmic artery by color Doppler imaging, corneal biomechanical parameters using a non-contact tonometer Corvis ST, and the corneal pulse (CP) signal using a non-contact ultrasonic technique. Statistically significantly lower mean values of normalized amplitudes of higher CP harmonics and changes in eight of the twelve corneal biomechanical parameters were observed in the rabbit group with lower arterial blood pressure and higher heart rate, intraocular pressure, and resistive index. The results of partial correlations showed that the CP signal energy and amplitude of its first harmonic correlate with the resistive index, diastolic and mean arterial pressures, whereas no statistically significant correlation was found between any of the CP parameters and intraocular pressure. Our pilot study indicates, for the first time, that non-contact and continuous measuring of corneal pulse allows indirectly assessing changes in cardiovascular activity when the confounding effect of intraocular pressure is eliminated.

The First Application of Nanoelectrochemotherapy in Feline Oral Malignant Melanoma Treatment-Case Study.

BACKGROUND: The aim of this study was to present the first domestic animal trial of nanosecond electroporation with chemotherapy and CO2 laser surgery. METHODS: sixteen-year-old domestic cat with diagnosed melanoma on oral cavity was the case used in the study. Firstly, CO2 laser surgery was used for the removal of most of tumor mass. Then nanoelectrochemotherapy with bleomycin was applied including appropriate margin of healthy tissue. A 15 ± 4 kV/cm × 15 ns pulsing protocol was employed with a total of 800 ± 100 pulses. Only one session of nanoelectrochemotherapy (nanoECT) was performed. RESULTS AND CONCLUSIONS: during the next two weeks areas of focal necrosis were replaced by the granulation tissue. Complete wound healing was observed four weeks after initial treatment. After 15 weeks post treatment, no local recurrence was apparent.

Corneal pulsation and biomechanics during induced ocular pulse. An ex-vivo pilot study.

The purpose of this study was to ascertain the relationships between the amplitude of the corneal pulse (CP) signal and the parameters of corneal biomechanics during ex-vivo intraocular pressure (IOP) elevation experiments on porcine eyes with artificially induced ocular pulse cycles. Two experiments were carried out using porcine eyes. In the first one, a selected eye globe was subjected to three IOP levels (15, 30 and 45 mmHg), where changes in physical ocular pulse amplitude were controlled by infusion/withdrawal volumes (ΔV). In the second experiment, six eyes were subjected to IOP from 15 mmHg to 45 mmHg in steps of 5 mmHg with a constant ΔV, where corneal deformation parameters were measured using Corvis ST. In both experiments, at each IOP, the CP and IOP signals were acquired synchronically using a non-contact ultrasonic distance sensor and a pressure transmitter, respectively. Based on the amplitudes of the CP and IOP signals ocular pulse based corneal rigidity index (OPCRI) was calculated. Results indicate positive correlations between ΔV and the physical ocular pulse amplitude, and between ΔV and the corneal pulse amplitude (both p < 0.001). OPCRI was found to increase with elevated IOP. Furthermore, IOP statistically significantly differentiated changes in OPCRI, the amplitudes of CP and IOP signals and in most of the corneal deformation parameters (p < 0.05). The partial correlation analysis, with IOP as a control variable, revealed a significant correlation between the length of the flattened cornea during the first applanation (A1L) and the corneal pulse amplitude (p = 0.002), and between A1L and OPCRI (p = 0.003). In conclusion, this study proved that natural corneal pulsations, detected with a non-contact ultrasonic technique, reflect pressure-volume dynamics and can potentially be utilized to assess stiffness of the cornea. The proposed new rigidity index could be a simple approach to estimating corneal rigidity.

Biological Characteristics and Osteogenic Differentiation of Ovine Bone Marrow Derived Mesenchymal Stem Cells Stimulated with FGF-2 and BMP-2.

Cell-based therapies using mesenchymal stem cells (MSCs) are a promising tool in bone tissue engineering. Bone regeneration with MSCs involves a series of molecular processes leading to the activation of the osteoinductive cascade supported by bioactive factors, including fibroblast growth factor-2 (FGF-2) and bone morphogenetic protein-2 (BMP-2). In this study, we examined the biological characteristics and osteogenic differentiation potential of sheep bone marrow MSCs (BM-MSCs) treated with 20 ng/mL of FGF-2 and 100 ng/mL BMP-2 in vitro. The biological properties of osteogenic-induced BM-MSCs were investigated by assessing their morphology, proliferation, phenotype, and cytokine secretory profile. The osteogenic differentiation was characterized by Alizarin Red S staining, immunofluorescent staining of osteocalcin and collagen type I, and expression levels of genetic markers of osteogenesis. The results demonstrated that BM-MSCs treated with FGF-2 and BMP-2 maintained their primary MSC properties and improved their osteogenic differentiation capacity, as confirmed by increased expression of osteocalcin and collagen type I and upregulation of osteogenic-related gene markers BMP-2, Runx2, osterix, collagen type I, osteocalcin, and osteopontin. Furthermore, sheep BM-MSCs produced a variety of bioactive factors involved in osteogenesis, and supplementation of the culture medium with FGF-2 and BMP-2 affected the secretome profile of the cells. The results suggest that sheep osteogenic-induced BM-MSCs may be used as a cellular therapy to study bone repair in the preclinical large animal model.