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.

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.

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.

Prevalence of common canine digestive problems compared with other health problems in teaching veterinary hospital, Faculty of Veterinary Medicine, Cairo University, Egypt.

AIM: The present study was conducted to ascertain the prevalence of common digestive problems compared to other health problems among dogs that were admitted to the teaching veterinary hospital, faculty of veterinary medicine, Cairo University, Egypt during 1 year period from January to December 2013. Also, study the effect of age, sex, breeds, and season on the distribution of digestive problems in dogs. MATERIALS AND METHODS: A total of 3864 dogs included 1488 apparently healthy (included 816 males and 672 females) and 2376 diseased dogs (included 1542 males and 834 females) were registered for age, sex, breed, and the main complaint from their owners. A complete history and detailed clinical examination of each case were applied to the aids of radiographic, ultrasonographic, and endoscopic examination tools. Fecal examination was applied for each admitted case. Rapid tests for parvovirus and canine distemper virus detection were also performed. RESULTS: A five digestive problems were commonly recorded including vomiting, diarrhea, concurrent vomiting with diarrhea, anorexia, and constipation with a prevalence (%) of 13.6, 19.1, 10.1, 13.1, and 0.5 respectively while that of dermatological, respiratory, urinary, neurological, cardiovascular, auditory, and ocular problems was 27.9, 10.5, 3.3, 0.84, 0.4, 0.25, and 0.17 (%) respectively. This prevalence was obtained on the basis of the diseased cases. Age and breed had a significant effect on the distribution of digestive problems in dogs (p<0.001). Gender had an effect on the distribution of digestive problems with significant (p≤0.01) while season had a non-significant effect (p>0.05) on the distribution of such problems. CONCLUSION: Digestive problems were the highest recorded problems among dogs, and this was the first records for such problems among dogs in Egypt. Age, gender, and breeds had a significant effect on the distribution of the digestive problems in dogs while season had a non-significant effect on the distribution of such problems. The present data enable veterinarians in Egypt to ascertain their needs for diagnostic tools and medication that must be present at any pet clinic.

Intrathecal Transplantation of Autologous Adherent Bone Marrow Cells Induces Functional Neurological Recovery in a Canine Model of Spinal Cord Injury.

Spinal cord injury (SCI) results in demyelination of surviving axons, loss of oligodendrocytes, and impairment of motor and sensory functions. We have developed a clinical strategy of cell therapy for SCI through the use of autologous bone marrow cells for transplantation to augment remyelination and enhance neurological repair. In a preclinical large mammalian model of SCI, experimental dogs were subjected to a clipping contusion of the spinal cord. Two weeks after the injury, GFP-labeled autologous minimally manipulated adherent bone marrow cells (ABMCs) were transplanted intrathecally to investigate the safety and efficacy of autologous ABMC therapy. The effects of ABMC transplantation in dogs with SCI were determined using functional neurological scoring, and the integration of ABMCs into the injured cords was determined using histopathological and immunohistochemical investigations and electron microscopic analyses of sections from control and transplanted spinal cords. Our data demonstrate the presence of GFP-labeled cells in the injured spinal cord for up to 16 weeks after transplantation in the subacute SCI stage. GFP-labeled cells homed to the site of injury and were detected around white matter tracts and surviving axons. ABMC therapy in the canine SCI model enhanced remyelination and augmented neural regeneration, resulting in improved neurological functions. Therefore, autologous ABMC therapy appears to be a safe and promising therapy for spinal cord injuries.

Autologous bone marrow-derived cell therapy combined with physical therapy induces functional improvement in chronic spinal cord injury patients.

Spinal cord injuries (SCI) cause sensory loss and motor paralysis. They are normally treated with physical therapy, but most patients fail to recover due to limited neural regeneration. Here we describe a strategy in which treatment with autologous adherent bone marrow cells is combined with physical therapy to improve motor and sensory functions in early stage chronic SCI patients. In a phase I/II controlled single-blind clinical trial (clinicaltrials.gov identifier: NCT00816803), 70 chronic cervical and thoracic SCI patients with injury durations of at least 12 months were treated with either intrathecal injection(s) of autologous adherent bone marrow cells combined with physical therapy or with physical therapy alone. Patients were evaluated with clinical and neurological examinations using the American Spinal Injury Association (ASIA) Impairment Scale (AIS), electrophysiological somatosensory-evoked potential, magnetic resonance imaging (MRI), and functional independence measurements. Chronic cervical and thoracic SCI patients (15 AIS A and 35 AIS B) treated with autologous adherent bone marrow cells combined with physical therapy showed functional improvements over patients in the control group (10 AIS A and 10 AIS B) treated with physical therapy alone, and there were no long-term cell therapy-related side effects. At 18 months posttreatment, 23 of the 50 cell therapy-treated cases (46%) showed sustained functional improvement. Compared to those patients with cervical injuries, a higher rate of functional improvement was achieved in thoracic SCI patients with shorter durations of injury and smaller cord lesions. Therefore, when combined with physical therapy, autologous adherent bone marrow cell therapy appears to be a safe and promising therapy for patients with chronic SCI of traumatic origin. Randomized controlled multicenter trials are warranted.