Effects of steroids on the morphology and proliferation of canine and equine mesenchymal stem cells of adipose origin — in vitro research

Disorders of the locomotive system, especially those occurring due to degenerative changes of the joints, are serious problems in daily veterinary medical practice. Steroid injections are the main way of treating these disorders. However, this approach brings usually only temporary effects of pain relief, and may cause many side effects. Alternative therapies focus on regeneration of damaged tissue using adult mesenchymal stem cells (MSCs). Since 2002, the great plasticity and immunomodulatory properties of MSCs isolated from adipose tissue (AdMSCs) have been used successfully in the treatment of degenerative joint diseases (DJD) of both dogs and horses. Possible simultaneous application of steroid therapy and stem cell transplantation could improve the commonly used clinical procedure. In this paper, the influence of the two steroid drugs (betamethasone and methylprednisolone) on AdMSCs was evaluated on the basis of morphology and proliferation rate. Both steroids positively influenced the viability and proliferation state of cells in a concentration of 0.01 mg/ml and 0.1 mg/ml, respectively. However, the concentration of 1 mg/ml had a cytotoxic effect. Moreover, the lower dosage of steroid drugs used in the experiment did not affect the morphology of cells and significantly increased cellular activity. In conclusion, our data demonstrate the stimulating effect of steroid drugs on cell morphology, proliferation rate and cytophysiological activity. These findings may influence the use of stem cells and steroids in applied regenerative veterinary medical practice in the future

The effects of the DNA methyltranfserases inhibitor 5-Azacitidine on ageing, oxidative stress and DNA methylation of adipose derived stem cells

Human adipose tissue is a great source of adult mesenchymal stem cells (MSCs) which are recognized from their ability to self‐renew and differentiation into multiple lineages. MSCs have promised a vast therapeutic potential in treatment many diseases including tissue injury and immune disorders. However, their regenerative potential profoundly depends on patients’ age. Age‐related deterioration of MSC is associated with cellular senescence mainly caused by increased DNA methylation status, accumulation of oxidative stress factors and mitochondria dysfunction. We found that DNA methyltransferase (DNMT) inhibitor i.e. 5‐Azacytidine (5‐AZA) reversed the aged phenotype of MSCs. Proliferation rate of cells cultured with 5‐AZA was increased while the accumulation of oxidative stress factors and DNA methylation status were decreased. Simultaneously the mRNA levels of TET proteins involved in demethylation process were elevated in those cells. Moreover, cells treated with 5‐AZA displayed reduced reactive oxygen species (ROS) accumulation, ameliorated superoxide dismutase activity and increased BCL‐2/BAX ratio in comparison to control group. Our results indicates that, treating MSCs with 5‐AZA can be justified therapeutic intervention, that can slow‐down and even reverse aged‐ related degenerative changes in those cells

Intracranial hematoma as the cause of headache after subarachnoid anesthesia for cesarean section – a case report

Background: Intracranial subdural hematoma is an exceptionally rare but life-threating complication of epidural and spinal anesthesia. The diagnosis is rather difficult because the initial symptoms mimic post-dural puncture headache. Case report: A 33-year-old primipara was admitted to the hospital at 38 weeks gestation for a cesarean section due to premature rupture of membranes and meconium stained amniotic fluid. During the procedure a single puncture between L2 and L3 vertebrae was made with the use of a 26-gauge, pencil-point needle. The amount of 2.8 ml of analgesic solution was administered in order to obtain subarachnoid analgesia at the level of Th4 and Th5 vertebrae. Postpartum recovery was uneventful for the first two days. On the third day the patient developed strong headache in the forehead area and tinnitus. An anesthesiologist diagnosed post-dural puncture headache (PDPH). The patient received 1g of Paracetamol every 6 hours intravenously, together with 3000 ml of crystalloid solution for 24 hours. As a result, the patient recovered and was discharged home with her infant. Five days later the patient presented at the neurology clinic because of strong and chronic temporal lobe headache. No other complaints were reported. Upon admission, the patient had a head CT, followed by an MRI examination, which revealed cranial hematomas localized bilaterally in the area of the frontal, temporal and parietal lobes, spreading from the cranial vault to the skull base. The width of the hematomas was: 3-4 mm on the left and 5-6 mm on the right side. Hematomas infiltrated the anterior part of the medial longitudinal fissure. Magnetic resonance angiography showed normal images of the arteries, veins, and the dural venous sinuses. No vascular malformations, which may be a source of intracranial hemorrhage, were found. Other tests showed normal results. Patient condition during hospitalization was stable. Conservative treatment was implemented, i.e. fluids administered intravenously, anti-edematous drugs, analgesic medications and bed rest. All pain complaints subsided and a control CT scan showed that hematomas evolved as expected i.e. their HU density decreased. About 6 weeks later the patient had a CT head scan, performed in outpatient settings, which showed complete absorption of extravasated blood. Conclusion: The presented case shows headaches in obstetric patients require thorough diagnostic examinations and appropriate management. In addition to the most typical PDPH, it may be the first sign of life-threatening intracranial pathology

Współistnienie ziarniniaka grzybiastego i przewlekłej białaczki limfocytowej B-komórkowej

The authors present a rare case of co-existence of mycosis fungoides of fairly rapid progress and chronic lymphocytic leukemia (B-CLL) in a 73-year-old man. The diagnostic and therapeutic difficulties resulted from highly variable presentations and the complex relationship between the two cancers are reported in the context of multidisciplinary approach to this type of disorder.W niniejszej pracy przedstawiono rzadki przypadek współistnienia ziarniniaka grzybiastego o dość szybkim przebiegu i przewlekłej białaczki limfocytowej B-komórkowej u mężczyzny w wieku 73 lat. Opisano trudnościdiagnostyczno-terapeutyczne ze względu na zróżnicowany obraz kliniczny zmian skórnych oraz złożony związek między dwoma nowotworami, uwzględniając kontekst wielodyscyplinarnego leczenia

Intraoperative parameters and postoperative follow-up of foam-based intraperitoneal chemotherapy (FBIC)

Background: For decades, intraperitoneal chemotherapy (IPC) has been delivered into the abdominal cavity as a liquid solution. Recently the concept of foam as a carrier-solution for IPC was suggested. This in-vivo swine study aims to evaluate the safety, intraoperative parameters, limitations and postoperative complications of foam-based intraperitoneal chemotherapy (FBIC).Methods: Three 65-day-old swine received FBIC with doxorubicin in a laparoscopy setting. Intraoperative parameters were monitored throughout the procedure and an extensive postoperative laboratory monitoring was conducted for 7 days. At day seven an autopsy was performed for further evaluation.Results: The insufflation of FBIC caused a temporary rise in blood pressure and a simultaneous drop in heart rate. Capnography detected a continuous increase in end-tital CO2 levels. A temporary drop of intraabdominal temperature was noted. Postoperative blood and serum laboratory results did not indicate any organ failure. No indication of intraperitoneal infections was noted and no structural tissue changes were visible in the autopsy.Discussion: The application of FBIC appears to be a feasible approach regarding intraoperative anesthesiology and postoperative surgical management. A lack of postoperative structural changes on the seventh day were a promising sign of safety and biocompatibility. Surgical reintervention would have been possible. To discuss a possible clinical application, further studies are required to investigate long-term safety, pharmacodynamics and the antitumoral potential of FBIC

The use of extracorporeal shock wave therapy in the treatment of orthopedic diseases in horses

This paper aims at presentation of advantages of extracorporeal shock wave therapy (ESWT), as a form of rehabilitation of the musculoskeletal system in horses. ESWT is a stream of acoustic waves generated outside the body. The equipment builds up pressure ranging from 10 to 100 MPa in the time from 30 to 120ns with short pulse duration equal to 5us. The appropriate parameters depend on energy level, pulse frequency, type of injury, type of treated tissue and depth of penetration. ESWT has an analgetic effect in the treated tissue, accelerates the healing process, increases collagen production, improves microcirculation and angiogenesis. Indications for the use of ESWT are insertional desmopathies, superficial and deep digital flexor tendonitis, metacarpal and metatarsal fractures, osteroarthritis and back pain. The number of treatments depends on the type of tissue and the size of the lesion. The tissue response to treatment is also very important. As a rule, 3 to 5 treatments are performed with a break of 5 to 7 days between sessions. The horse during treatment should have limited movement and orthopedic control is very important

Chondrogenic potential of canine articular cartilage derived cells (cACCs)

In the present paper, the potential of canine articular cartilage-derived cells (cACCs) for chondrogenic differentiation was evaluated. The effectiveness of cACCs’ lineage commitment was analyzed after 14 days of culture in chondorgenic and non-chondrogenic conditions. Formation of proteoglycan-rich extracellular matrix was assessed using histochemical staining – Alcian Blue and Safranin-O, while elemental composition was determined by means of SEM-EDX. Additionally, ultrastructure of cACCs was evaluated using TEM. The expression of genes involved in chondrogenesis was monitored with quantitative Real Time PCR. Results obtained indicate that the potential of cACCs for cartilagous extracellular matrix formation may be maintained only in chondrogenic cultures. The formation of specific chondro-nodules was not observed in a non-chondrogenic culture environment. The analysis of cACCs’ ultrastructure, both in non-chondrogenic and chondrogenic cultures, revealed well-developed rough endoplasmatic reticulum and presence of mitochondria. The cACCs in chondrogenic medium shed an increased number of microvesicles. Furthermore, it was shown that the extracellular matrix of cACCs in chondrogenic cultures is rich in potassium and molybdenum. Additionally, it was determined that gene expression of collagen type II, aggrecan and SOX-9 was significantly increased during chondrogenic differentiation of cACCs. Results obtained indicate that the culture environment may significantly influence the cartilage phenotype of cACCs during long term culture

Central venous catheters and complications

Stały dostęp dożylny ma ogromne znaczenie w leczeniu chorych onkologicznych, którzy poddawani są długotrwałej chemioterapii. Współczesna medycyna nie mogłaby istnieć bez stałego i pewnego dostępu dożylnego. W wielu oddziałach stosuje się rutynowo centralne dostępy żylne typu „port”, które pozwalają choremu na swobodne poruszanie się bez obawy o przypadkowe usunięcie cewnika. Port dożylny składa się ze zbiorniczka z silikonową membraną, która może być wielokrotnie nakłuwana oraz cienkościennego cewnika wewnątrznaczyniowego. Port stanowi niezaprzeczalne udogodnienie dla chorych i może spełniać swoją rolę przez wiele miesięcy, a nawet lat. Szczególnie w onkologii konieczność zapewnienia takiego trwałego dostępu naczyniowego jest istotnym elementem prowadzenia terapii, ze względu na silnie drażniące działanie wielu cytostatyków. Celem pracy jest przedstawienie obecnych standardów oraz zaleceń zastosowania portu dożylnego, a także najczęściej występujących powikłań, wczesnych i późnych związanych z obecnością centralnego cewnika żylnego. Na podstawie przeglądupiśmiennictwa autorzy przedstawili ryzyko wystąpienia typowych powikłań dla kaniulacji żył centralnych, mechanizm i czynniki wpływające na przebieg powikłań związanych z użytkowaniem portów dożylnych. Powikłania są nieodłączną częścią wszelkich zabiegów medycznych, dlatego niezbędne jest ich poznanie oraz opanowanie odpowiednich metod postępowania terapeutycznego i profilaktyki. Zastosowanie portów zyskuje coraz większą popularność w polskiej onkologii. W związku z tym lekarze powinni szczególnie znać zalety i typowe powikłania dotyczące portów naczyniowych. Większość powikłań wynika z niedostosowania się do procedur, braku odpowiedniego nadzoru, niedbałości lub po prostu niedostatecznej wiedzy.Permanent venous line access is of a great importance in the treatment of oncological patients who are undergoing long-term chemotherapy. Modern medicine could not exist without a regular and reliable venous line access. In many departments central venous line (venous port systems), which allows a patient to move freely without fear of accidental removal of a catheter are used routinely. Venous port system consists of a reservoir compartment with a silicone membrane - septum, which can be repeatedly punctured, and a thin-walled intravascular catheter. A port is an undeniable convenience for patients and can fulfill its role for many months or even years. Especially in oncology the need to ensure permanent vascular line access is an essential part of the therapy due to severe irritation caused by many cytostatics. The aim of this study is to present the current standards and guidelines concerning intravenous port system and the most common early and late complications caused by the presence of central venous catheter. Based on a review of the bibliographical data, the authors presented therisk of common complications of central venous cannulation, the mechanism and the factors influencing the complications associated with the use of venous ports. Complications are an integral part of any medical treatment, therefore it is essential to understand their mechanism and learn appropriate methods of treatment and prevention. The use of ports is becoming increasingly popular in Polish oncology. Therefore, physicians should be familiar with advantages and common complications of the use of venous ports. Most of the complications result from the failure to comply with procedures, lack of adequate supervision, negligence or simply insufficient knowledge

Immunohistochemical Analysis of Renal Solid and Papillary-Ductal Carcinoma in Dogs

The paper includes description of neoplastic lesions in canine kidneys, sampled during surgery or post mortem. The tumours were subjected to morphological and immunohistochemical analysis using antibodies specific for c-kit, cytokeratin AE1/AE3, CD10, vimentin, Carcinoembryonic Antigen (CAE), Ki-67. The obtained results permitted to specify an accurate histopathological diagnosis

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