Relationship Between Arterial and Intraosseous Pressure in African Grey Parrots (Psittacus erithacus).

The aim of this study was to determine the relationship between arterial pressure and intraosseous (IO) pressure obtained by conventional IO needle in African grey parrots (Psittacus erithacus). Twelve healthy African grey parrots with mean body weight 465.25 g were anesthetized in an induction chamber with 5% isoflurane and 100% oxygen, followed by intubation and connection to a pressure-limited intermittent-flow ventilator. A 24-gauge catheter was placed into the deep radial artery in one wing and a 22-gauge needle was placed into the distal ulnar medullary cavity in the contralateral wing. Both cannulas were connected to a 9-cm noncompliant tube and then linked to a disposable pressure sensor. Pressures and waveforms from both cannulas were recorded for 10 minutes while isoflurane was maintained at 3%. The results showed that the arterial pressure values were normally distributed (P < .05), whereas the IO pressure values were not (P > .05). Arterial and IO pressure were not correlated (P > 0.05), but IO pressure and heart rate were positively correlated (Spearman's rank correlation coefficient, 0.58; P < .05). Multiple characteristics of the IO waveforms were observed, including distinguishable continuous waveforms (n = 8), a single wave crest (n = 4), respiratory variation (n = 7), and a dicrotic notch (n = 3), and IO waveforms were appearing at a similar or delayed point relative to the related arterial waveforms (n = 8). Although IO pressure was not correlated with arterial pressure in the African grey parrots under the study conditions, the positive correlation between IO pressure and heart rate and the IO waveform characteristics suggest that the inflow arterioles likely account for the retained arterial waveform radiating into the IO space. These findings are consistent with those from all previously tested mammals. Therefore, insights into potential future improvements of the measuring techniques and protocols can be reached by thoroughly reviewing the current knowledge on IO pressure in mammals.

Effects of Focal Radiation on [18 F]-Fluoro-D-Glucose Positron Emission Tomography in the Brains of Miniature Pigs: Preliminary Findings on Local Metabolism.

OBJECTIVES: It would be a medically important advance if durable and focal neuromodulation of the brain could be delivered noninvasively and without ablation. This ongoing study seeks to elucidate the effects of precisely delivered ionizing radiation upon focal brain metabolism and the corresponding cellular integrity at that target. We hypothesize that focally delivered ionizing radiation to the brain can yield focal metabolic changes without lesioning the brain in the process. MATERIALS AND METHODS: We used stereotactic radiosurgery to deliver doses from 10 Gy to 120 Gy to the left primary motor cortex (M1) of Lee Sung miniature pigs (n = 8). One additional animal served as a nonirradiated control. We used positron emission tomography-computed tomography (PET-CT) to quantify radiation dose-dependent effects by calculating the ratio of standard uptake values (SUV) of 2-deoxy-2-[18 F]-fluoro-D-glucose (18 F-FDG) between the radiated (left) and irradiated (right) hemispheres across nine months. RESULTS: We found that the FDG-PET SUV ratio at the targeted M1 was significantly lowered from the pre-radiation baseline measurements for animals receiving 60 Gy or higher, with the effect persisting at nine months after radiosurgery. Only at 120 Gy was a lesion suggesting ablation visible at the M1 target. Animals treated at 60-100 Gy showed a reduced signal in the absence of an identifiable lesion, a result consistent with the occurrence of neuromodulation. CONCLUSION: Focal, noninvasive, and durable changes in brain activity can be induced without a magnetic resonance imaging (MRI)-visible lesion, a result that may be consistent with the occurrence of neuromodulation. This approach may provide new venues for the investigation of neuromodulatory treatments for disorders involving dysfunctional brain circuits. Postmortem pathological analysis is needed to elucidate whether there have been morphological changes not detected by MRI.

Evaluation of endoscopic salpingectomy for sterilization of female Formosan macaques (Macaca cyclopis).

We evaluated the safety and postsurgical outcomes of endoscopic salpingectomy for sterilization of female Formosan macaques (Macaca cyclopis) as a method of population control. Nineteen adult female Formosan macaques were included in our study. The fallopian tubes of each anesthetized macaque were cauterized and excised endoscopically using a 2.7-mm rigid endoscope system. We recorded the complications encountered, and objectively scored the amount of hemorrhage throughout the procedure. Postoperative ovarian function was evaluated by monitoring the serum levels of sex hormones in ten of the macaques for two ovarian cycles following the salpingectomy. Two to 13 months later, eight of the 19 macaques underwent laparoscopy for the objective evaluation of inflammation at the surgical sites on the fallopian tubes. No major anesthetic- or surgical-associated complications were observed in any of the macaques. The hormonal evaluation showed cyclic ovarian function after salpingectomy in all of the ten macaques examined, and the parameters were comparable to those of other macaque species. The long-term postoperative level of inflammation at the surgical site was minimal to low, and was lower than that reported for other tubal occlusion techniques used in macaques. The use of a 2.7-mm rigid endoscope for salpingectomy in macaques is safe and efficient, with fewer postoperative complications than comparable sterilization techniques.

Long-term Management of Thymic Lymphoma in a Java Sparrow (Lonchura oryzivora).

A 4-year-old Java sparrow (Lonchura oryzivora) was diagnosed with thymic lymphoma based on the results of histopathologic evaluation and immunohistochemical staining of a Tru-cut biopsy sample. The bird was treated with chlorambucil (2 mg/kg PO) twice per week for the first 18 weeks, and the mass size, activity level, appetite, bowel function, and body weight were monitored weekly. At week 19, the bird became ill and anorexic, and the chlorambucil treatment was discontinued. The neoplasm grew rapidly during weeks 20 and 21, and the chlorambucil treatment was resumed at week 23. At week 28, the bird's overall condition worsened and the chlorambucil treatment was discontinued because no additional reduction in mass size had occurred. At week 29, prednisolone (2.2 mg/kg) was administered orally every 12 h for 31 weeks. At week 61, the patient became acutely ill, exhibiting anorexia and a decreased level of physical activity, and the owner elected to euthanatize the bird. This is the first report of long-term management of thymic lymphoma in a Java sparrow.

Natural A(H1N1)pdm09 influenza virus infection case in a pet ferret in Taiwan.

Ferrets have demonstrated high susceptibility to the influenza virus. This study discusses a natural 2009 pandemic influenza A (H1N1) (A(H1N1)pdm09) virus infection in a pet ferret (Mustela putorius furo) identified in Taiwan in 2013. The ferret was in close contact with family members who had recently experienced an influenza-like illness (ILI). The ferret nasal swab showed positive results for influenza A virus using one-step RT-PCR. The virus was isolated and the phylogenetic analysis indicated that all of the eight segmented genes were closely related to the human A(H1N1)pdm09 virus linage isolated in Taiwan. This study may provide a perspective view on natural influenza A virus transmission from the local human population into pet ferrets.

Octanoic acid promotes clearance of antibiotic-tolerant cells and eradicates biofilms of Staphylococcus aureus isolated from recurrent bovine mastitis.

Antibiotic therapy is the primary treatment for bovine mastitis, but the drawbacks of this strategy include poor cure rate and economic losses from the need to discard milk with antibiotic residues. Unfortunately, few other treatment options are currently available for mastitis. Failure of antibiotic treatments is often attributed to formation of bacterial biofilms and abscesses in the mammary gland tissue, which lead to chronic infections that are difficult to eradicate and drive recurrent disease. A major mastitis-causing pathogen (MCP) associated with biofilms in bovine mastitis is Staphylococcus aureus. In this study, we demonstrate that octanoic acid has broad-spectrum microbicidal activity against MCPs and effectively inhibits S. aureus biofilm formation in milk (>50% inhibition at 3.13 mM). Octanoic acid effectively clears biofilms (95% eradication at 1X minimum bactericidal concentration, MBC) and infrequently induces S. aureus small colony variants (SCVs) that may cause recurrent mastitis. Additionally, octanoic acid rapidly kills persistent biofilm cells and cells with antibiotic tolerance (within 4 h). In contrast, antibiotics treated at >100X MBC cannot eradicate biofilms but do induce SCVs and antibiotic-tolerant cells. These effects may accelerate the transition from biofilm to chronic infection. Thus, octanoic acid exhibits bactericidal action against S. aureus biofilms, and it is less likely than antibiotic therapy to induce persistent cells and pathogen tolerance. Moreover, octanoic acid acts additively with antibiotics against S. aureus, and it attenuates tetracycline-induced virulence factor gene expression in S. aureus cells. According to these data, octanoic acid may prevent the pathological progression of bovine mastitis and offer a new strategy for treating the condition.

Observation of reproductive cycle of female yellow-margined box turtle (Cuora flavomarginata) using radiography and ultrasonography.

This study presents a combination of radiography and ultrasonography to observe the reproductive cycle of 24 captive female yellow-margined box turtles in Taiwan. Radiography was mainly used to monitor clutch size, whereas ultrasonography was applied to detect changes in the follicles throughout the year. The observation of the 24 female turtles was performed from April 2007 to June 2008. Their average carapace length was 16.62 ± 1.66 cm and their average body weight was 812 ± 164.98 g. The mean clutch size was three (87 eggs/29 clutches) and the reproductive frequency was 95.83% (23/24). Double clutches were detected in 79.2%, and 20.8% had single clutches. Ovulation occurred from March through August, and the average follicular diameter was 2.16 ± 0.18 cm. Follicles entered the latent period in October (at 1.54 ± 0.26 cm), and vitellogenesis of the next reproductive cycle began in November. Using radiography, the eggshell could be detected on the ninth day after ovulation. The average period of the single clutch group was 6.9 weeks (range 5.1-8.5 weeks). In the double clutch group, the average period of the first clutch was 5.5 weeks (range 4-7.8 weeks) and that of the second clutch was 5.2 weeks (range 4-7.8 weeks). This study has advanced the understanding of reproductive physiology of yellow-margined box turtle and established a valuable and practical model for comparative study of the reproductive physiology of other chelonians.

A robotized handheld smart tool for orthopedic surgery.

BACKGROUND: Handheld surgical robots offer functionalities, such as active guidance, tremor suppression and force reflection, for surgeons to enhance their skill in manipulating surgical tools during medical intervention. In orthopedic surgery, the robot additionally has to offer sufficient rigidity and power for bone machining. The size and weight of the mechanical design, together with the control behaviour associated with involuntary hand motion, navigation and reflected force to the human, all influence the overall performance of an orthopedic handheld robot. METHODS: The paper proposes a miniature and compact design for an embedded robot, which is a similar weight as a handpiece. Then, a shared controller is proposed to address the coupling among involuntary and voluntary hand motions, robot navigation, tool feedback forces and force artefacts from actuation. RESULTS: The handheld robot is able to stabilize the drill positioning by removing involuntary tremors as well as reduce force artefacts from motor actuation in experiments involving pedicle tunnelling on a porcine spine. CONCLUSION: The paper has successfully realized a compact handheld orthopedic robot which provides high performance of usability, tremor suppression and force reflection for bone drilling.

Observations on the female reproductive cycles of captive Asian yellow pond turtles (Mauremys mutica) with radiography and ultrasonography.

The annual reproductive cycle of 27 female Mauremys mutica was observed by radiography and ultrasonography from April 2006 to August 2007. Radiography was used to monitor clutch size and ultrasonography was used to monitor changes in the ovarian follicles. The follicles started to enlarge in September and became preovulatory in January. The mean maximum follicle diameter of ovulation was 18.30+/-1.44 mm, and ovulation occurred from March through August. Eggs were laid between April and August. Turtles entered latent period in early August and the maximum follicular size was at a low of 13.22+/-2.36 mm in late September. The vitellogenesis of the next reproductive cycle began in October. The 24 adult females laid 56 clutches containing a total of 227 eggs. Average clutch size was 4.05 eggs (range 1-8) and there were 2.33 clutches (range 1-4) per female. Egg shell images were first observed on the sixth or seventh day after ovulation. The oviductal period averaged 6.9 weeks (range 2-16 weeks) on the first clutch, 3.4 weeks (range 2-8 weeks) on the second, and 2.75 weeks (range 2-6 weeks) for the third. Radiography and ultrasonography are non-invasive and convenient methods to evaluate the reproductive cycle of female M. mutica. They should be applicable to other turtles and should greatly enhance knowledge of reproductive physiology.

BLOOD LEAD AND ZINC LEVELS AND THEIR IMPACT ON HEALTH OF FREE-LIVING SMALL CARNIVORES IN TAIWAN, REPUBLIC OF CHINA.

Lead and zinc are recognized as the most widespread trace metals in nature and can, at high levels, compromise the health of wildlife and their habitat. Because of their position in a higher trophic level, wild carnivores can be valuable biological indicator species of trace-metal contamination in the environment. We assessed blood lead and zinc concentrations of four small carnivore species native to Taiwan, the small Indian civet (Viverricula indica), the masked palm civet (Paguma larvata), the ferret badger (Melogale moschata), and the crab-eating mongoose (Herpestes urva), from urban and rural areas (Yangmingshan National Park, Xiuguluan River bank, and Da-an River bank). Blood samples were acquired from the anterior vena cava under general anesthesia, and lead and zinc concentrations, hematology, and serum biochemistry results were then obtained. Blood lead levels were significantly higher in ferret badgers in the Yangmingshan area. Although lead concentrations were comparable with those in humans and cats with lead toxicosis, there was no hematological or biochemical evidence that animal health was compromised. Blood zinc levels were within an acceptable range in all four species tested. Overall, we found significant differences in blood lead and zinc levels among four species of carnivores living in areas with different levels of land development in Taiwan. Anthropogenic pollution, mining history, and volcanic activities in Yangmingshan National Park may contribute to significantly high blood lead levels in ferret badgers in this area. Our results provided information about the potential impact of land development on wildlife and may be beneficial to wildlife conservation, public health, and environmental health in Taiwan.

Light Microscopic and Ultrastructural Characteristics of Heterophil Toxicity and Left-shifting in Green Sea Turtles (Chelonia mydas) from Taiwan.

Heterophil toxic change (TC) and left-shifting (LS) are widely used as indicators of accelerated granulopoiesis. However, the ultrastructure of heterophil TC and LS in sea turtles remain poorly understood. This study aimed to describe the ultrastructural characteristics of sea turtle TC and LS heterophils, compare the staining quality of accessible staining methods, and provide a better understanding of the clinical applications and limitations of heterophil TC and LS examinations. Blood samples were collected from 21 rescued sea turtles from January 2017 to September 2018. Morphologic (n = 22) and ultrastructural (n = 15) examination of TC and LS heterophils were performed, and the qualities of three staining methods (Wright-Giemsa stain, Diff-Quik stain and Liu's stain) were analyzed to diagnose TC and LS heterophils. In addition, the diagnostic values of TC and LS heterophils were examined. Diff-Quik stain was significantly inferior in the assessment of heterophil TC and/or LS comparing to the Wright-Giemsa stain and Liu's stain (Mann-Whitney test, P < 0.001). Microscopic examinations of heterophil TC and/or LS were comparable to transmission electron microscopy examinations (Cohen's kappa coefficient, κ = 1). The correlation between the presence of heterophil TC and/or LS and clinical inflammatory state was weak (Spearman's rank correlation coefficient, rs = 0.171, p = 0.445). In conclusion, this is the first study to describe the ultrastructural characteristics of reptile TC and LS heterophils. Wright-Giemsa stain and Liu's stain were suitable staining methods for the microscopic observations of TC and LS heterophil in sea turtles. Given the poor correlation between TC and/or LS and clinical findings, TC and LS are not a suitable diagnostic indicator of green sea turtles' inflammation status.

Morphologic and cytochemical characteristics of Chinese striped-necked [corrected] turtle (Ocadia sinensis) blood cells and their hematologic and plasma biochemical reference values.

Hematologic analyses are useful for the monitoring of animal health and diseases and for the differentiation of physiologic processes for clinicians and conservationists. In order to establish hematology reference values for the Chinese striped-necked [corrected] turtle (Ocadia sinensis) and to produce an accurate baseline of clinical laboratory data for O. sinensis with regard to sex and season, 50 (24 males and 26 females) adult captive individuals of O. sinensis were studied. Blood samples from the jugular veins of the turtles were collected in January, April, June, and November. Data were analyzed using repeated measures analysis of variance for significant (P < 0.05) variation by sex, season, and the interaction between sex and season. Significant sex differences were observed for the parameters of packed cell volume, eosinophil count, heterophils and monocytes ratio, total protein, albumin, uric acid, aspartate aminotransferase, alanine aminotransferase, triglycerides, cholesterol, and alkaline phosphatase. Marked seasonal variation was noted in all parameters except mean cell hemoglobin, monocytes and heterophils ratio, and creatinine. Differences between sexes and seasons were primarily associated with the reproductive cycle. Heterophils had a strong positive reaction and eosinophils had a moderate positive reaction to benzidine peroxidase stain. Thrombocytes had a positive reaction to periodic acid-Schiff stain. Surface morphologic study using scanning electron microscopy of blood cells showed that white blood cells of O. sinensis had no distinctive surface characteristics.

Growth and Behavior of Congenitally Anophthalmic Lee-Sung Pigs.

Circadian rhythm is usually regulated by the environmental light-dark cycle. Congenitally anophthalmic miniature pigs provide a valuable model for the study of factors affecting circadian rhythms in the absence of visual exposure to the light-dark cycle. This study investigated the growth and daily behavior patterns of Lee-Sung pigs with congenital anophthalmia. Growth in 5 Lee-Sung pigs (LSP) with congenital anophthalmia (LSP-A) and 10 normally developed pigs (LSP-N) was assessed when they were 1 through 6 mo old. Behavioral studies using digital video recording were completed in 6 sexually mature LSP (3 LSP-A and 3 LSP-N). MRI showed that LSP-A lose their vision because of a lack of retinal input and optic chiasm development. LSP-N and LSP-A did not differ in body weight or size at 2, 4, and 6 mo of age. Behavior and activity pattern studies showed that both LSP-A and LSP-N were active mainly during daylight, but LSP-A spent significantly more time exploring their environment during the day (28%) and night (10%) than did LSP-N. This study revealed that growth performance was similar between LSP-A and normal pigs, but their behavior and activity patterns differed. LSP-A showed circadian rhythm abnormalities similar to those in blind humans. This study provides basic data on LSP-A as a model for studying compensatory cross-modal brain plasticity and hormone regulation in the absence of retinal input is deficient and for understanding the role of circadian rhythm regulation.

Development of a novel biochip for rapid multiplex detection of seven mastitis-causing pathogens in bovine milk samples.

To efficiently prevent and treat bovine mastitis and minimize its effect on the dairy industry, a sensitive, rapid, and specific test is required for identifying the mastitis-causing pathogens. In this study, a biochip capable of detecting 7 common species of mastitis-causing pathogens, including Corynebacterium bovis, Mycoplasma bovis, Staphylococcus aureus, and the Streptococcus spp. S. agalactiae, S. bovis, S. dysgalactiae, and S. uberis, within 6 hr was developed. The technique is based on DNA amplification of genes specific to the target pathogens and consists of 4 basic steps: DNA extraction of bacteria, polymerase chain reaction, DNA hybridization, and colorimetric reaction. To examine the accuracy and specificity of this biochip, a preliminary test with 82 random quarter milk samples were analyzed and compared with results from conventional microbiological methods conducted simultaneously. Results from all but 1 sample analyzed by the biochip were in agreement with those analyzed by bacteriology. The biochip could be a feasible tool for rapidly diagnosing mastitis-causing pathogens in milk and providing information for a more effective treatment to cure mastitis.

Hematologic, Plasma Biochemical, Protein Electrophoretic, and Total Solid Values of Captive Oriental Turtle Doves (Streptopelia orientalis).

I-Ting Tsai, Chau-Hwa Chi, and Pin-Huan Yu (2018) As part of ongoing studies on the blood profiles of Taiwanese avian species, hematology, plasma biochemistry, and serum protein electrophoresis values and hand-held refractometer readings of total solids were determined in Oriental turtle doves (Streptopelia orientalis). Blood samples were collected and analyzed from 44 healthy captive adult and sub-adult Oriental turtle doves. There were no significant gender differences in any of the parameters studied. However, adults had significantly higher values of total protein, albumin, globulin, cholesterol, sodium, chloride, and alpha2 globulin, while sub- adults had higher values of phosphorus and potassium. The refractometer readings were highly correlated with values of total protein obtained by the biuret method. The results of the current study provide valuable data for clinically evaluating Oriental turtle doves and will be beneficial for the conservation of this species.

Comparison of three computed tomographic angiography protocols to assess diameters of major arteries in African grey parrots (Psittacus erithacus).

OBJECTIVE To evaluate 3 contrast medium infusion (CMI) protocols for CT angiography (CTA) and measurement of major artery diameters in African grey parrots (Psittacus erithacus). ANIMALS 9 African grey parrots with no detectable cardiovascular disease. PROCEDURES Each bird was anesthetized and underwent placement of an IV catheter in the left basilic vein and 16-slice CTA scanning (started at peak aortic enhancement) with each of 3 CMI protocols at ≥ 1-month intervals. Protocol 1 involved catheter flushing with saline (0.9% NaCl) solution and IV infusion of iopamidol (2 mL) followed by saline solution (0.2 mL; total infused volume, 5 mL). Protocol 2 involved IV infusion of iopamidol (2 mL) followed by saline solution (0.4 mL; total infused volume, 2.4 mL). Protocol 3 involved catheter flushing with saline solution and IV administration of iopamidol (2 mL; total infused volume, 4.8 mL). The diameters of 6 major arteries were measured by 2 observers, and intra- and interobserver agreement, time-enhancement variables, and patient factors affecting contrast medium enhancement were assessed. RESULTS Among the 3 CMI protocols, CTA-derived arterial diameters differed significantly. Measurements obtained with protocol 2 were significantly larger than those obtained with the other protocols. Uniformity of the time-enhancement variables differed among CMI protocols. Patient factors had nonsignificant effects on contrast medium enhancement. CONCLUSIONS AND CLINICAL RELEVANCE Of the CMI protocols assessed, a 2-phase CMI protocol with a post-CMI saline solution flush was the most reliable for CTA-derived measurements of the major thoracic and abdominal arteries in African grey parrots. However, further technique modification is needed.

Progressive coxa vara by eccentric growth tethering in immature pigs.

The present study tested progressive coxa vara by eccentric growth tethering that might be used to correct coxa valga in cerebral palsy. Eight young pigs received screw fixation at inferior portion of right femoral head at age 4 months and were killed at age 7.25 months for bilateral femurs for comparison. The neck-shaft angle at the tethered side was significantly less than that at the control side (129.8 vs. 138.3 degrees , P<0.05). Histological study showed bony bar formation. Eccentric growth tethering by one screw resulted in a reduction of neck-shaft angle by 8.5 degrees and shortening of femoral length by 4%.

Simultaneous activation and inhibition of autophagy sensitizes cancer cells to chemotherapy.

While combined chemotherapy (CT) with an autophagy inducer and an autophagy inhibitor appears paradoxical, it may provide a more effective perturbation of autophagy pathways. We used two dissimilar cell lines to test the hypothesis that autophagy is the common denominator of cell fate after CT. HA22T cells are characterized by CT-induced apoptosis and use autophagy to prevent cell death, while Huh7.5.1 cells exhibit sustained autophagic morphology after CT. Combined CT and rapamycin treatment resulted in a better combination index (CI) in Huh7.5.1 cells than combined CT and chloroquine, while the reverse was true in HA22T cells. The combination of 3 drugs (triplet drug treatment) had the best CI. After triplet drug treatment, HA22T cells switched from protective autophagy to mitochondrial membrane permeabilization and endoplasmic reticulum stress response-induced apoptosis, while Huh7.5.1 cells intensified autophagic lethality. Most importantly, both cell lines showed activation of Akt after CT, while the triplet combination blocked Akt activation through inhibition of phospholipid lipase D activity. This novel finding warrants further investigation as a broad chemosensitization strategy.

Anti-tumor effect of in vivo IL-2 and GM-CSF electrogene therapy in murine hepatoma model.

BACKGROUND: We evaluated the effect of in vivo electrogene therapy (EGT), a newly-developed gene transfer method using electroporation on the induction of anti-cancer immunity. MATERIALS AND METHODS: The in vivo EGT was carried out by direct injection of plasmid DNAs encoding mouse interleukin-2 (IL-2) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in a subcutaneous murine hepatoma model of 1MEA.7R.1 cells. Six electric pulses were generated in situ from a square-wave electroporator fitted with a circular, six-needle electrode array. 1MEA.7R1 cells in vitro were modified to secret IL-2 (1MEA.7R.1/IL-2 cells). RESULTS: The 1MEA.7R.1/IL-2 cells had a similar cell doubling-time as their parent cells but showed a much slower growth rate on Balb/C mice. One, or 3 rounds of single gene EGT with IL-2 gene showed a dose-responsive effect of growth retardation. Co-administration of 3 rounds of IL-2/GM-CSF double genes EGT had a stronger growth inhibition effect than 3 rounds of IL-2 single gene EGT. Three rounds of IL-2/GM-CSF EGT rendered the tumor to a growth rate of stably transfected 1MEA.7R.1/IL-2 cells. Seven rounds of IL-2/GM-CSF EGT markedly inhibited the tumor growth. Reverse transciptase-polymerase chain reaction confirmed the expression of IL-2, GM-CSF and interferon-gamma within treated tumors. Systemic inhibitory effects can be demonstrated from tumor-re-challenged experiments on mice which received 3 rounds of double-gene EGT. The T cell proliferation assay revealed an increased T cell proliferation in double-gene EGT-treated mice. CONCLUSION: This experiment showed that partial systemic immunity can be provoked by IL-2/GM-CSF double-gene EGT. These findings suggest that our immuno-gene therapy protocol has the potential for future clinical applications.

Fibrin glue mixed with platelet-rich fibrin as a scaffold seeded with dental bud cells for tooth regeneration.

Odontogenesis is a complex process with a series of epithelial-mesenchymal interactions and odontogenic molecular cascades. In tissue engineering of teeth from stem cells, platelet-rich fibrin (PRF), which is rich in growth factors and cytokines, may improve regeneration. Accordingly, PRF was added into fibrin glue to enrich the microenvironment with growth factors. Unerupted second molar tooth buds were harvested from miniature swine and cultured in vitro for 3 weeks to obtain dental bud cells (DBCs). Whole blood was collected for the preparation of PRF and fibrin glue before surgery. DBCs were suspended in fibrin glue and then enclosed with PRF, and the DBC-fibrin glue-PRF composite was autografted back into the original alveolar sockets. Radiographic and histological examinations were used to identify the regenerated tooth structure 36 weeks after implantation. Immunohistochemical staining was used to detect proteins specific to tooth regeneration. One pig developed a complete tooth with crown, root, pulp, enamel, dentin, odontoblast, cementum, blood vessels, and periodontal ligaments in indiscriminate shape. Another animal had an unerupted tooth that expressed cytokeratin 14, dentin matrix protein-1, vascular endothelial growth factor, and osteopontin. This study demonstrated, using autogenic cell transplantation in a porcine model, that DBCs seeded into fibrin glue-PRF could regenerate a complete tooth.