Identification of sex in Carica papaya L. using RAPD markers

Brazil is currently the worlds largest producer of papaya (Carica papayaL.), producing fruits for both the domestic market and export. Only fruits from hermaphrodite plants are marketed because they have the necessary commercial characteristics, i.e. they are pear-shaped and have thicker flesh and a smaller internal cavity. Increased papaya yield has been limited mainly by the ratio of female to hermaphrodite (1: 2) plants normally occurring in orchards. This ratio causes great losses to papaya producers and the identification of the sex of seedlings during the nursery stage would be an important advance. In our study random amplified polymorphic DNA (RAPD) analysis was used to differentiate between the sexual forms of three commercial C. papaya cultivars belonging to the Solo group. RAPD assays using the BC210 primer were able to detect hermaphrodites in all of the cultivars tested. The BC210₄₃₈molecular marker was much better at papaya sex differentiation than other markers described in the literature. This revised version was published online in August 2006 with corrections to the Cover Date.     

Clinical evaluation of random skin flaps based on the subdermal plexus secured with sutures or sutures and cyanoacrylate adhesive for reconstructive surgery in dogs

OBJECTIVE: To evaluate the use of subdermal plexus skin flaps for closing defects after excision of cutaneous and subcutaneous tumors in dogs and to compare outcome of flaps secured with sutures and those secured with butyl-cyanoacrylate and intermittent sutures. STUDY DESIGN: Clinical study. ANIMALS: Fifteen dogs. METHODS: After excision of cutaneous or subcutaneous tumors the skin defect was reconstructed by random flaps based on the subdermal plexus. Flap skin edges were apposed with simple interrupted 4-0 monofilament nylon sutures (group 1; 5 dogs) or nylon sutures alternated with butyl-cyanoacrylate adhesive (group 2; 10 dogs). Flaps were evaluated every 48 hours when bandages were changed, until complete healing. RESULTS: Random flaps based on the subdermal plexus were effectively used to close wound defects; mean flap survival was 89%. Partial flap necrosis occurred in 4 dogs. Wound margins apposed with butyl-cyanoacrylate had thinner and more esthetic scars than sutured margins. CONCLUSION: Random flaps based on the subdermal plexus proved to be versatile for covering limb wounds after excision of cutaneous or subcutaneous tumors. Mean survival rate was comparable to that reported for axial pattern flaps. Butyl-cyanoacrylate adhesive was easy to apply, allowed accurate margin apposition with good cosmetic outcome and reduced sutures needed. CLINICAL RELEVANCE: Cyanoacrylate adhesive should be considered in lieu of suture closure to secure random skin flaps based on the subdermal plexus in dogs.     

Serum vascular endothelial growth factor in dogs with soft tissue sarcomas

This work aimed to evaluate serum vascular endothelial growth factor (VEGF) in 25 dogs with soft tissue sarcoma, and in 30 healthy dogs. Blood was collected once time from the control animals and three times, in the same way, from animals with sarcoma. Blood count was performed in the blood collected, and serum VEGF was measured by enzyme‐linked immunosorbent assay quantitative method. Serum VEGF in control animals was similar to patients with soft tissue sarcoma. There was a reduction in serum VEGF after the sarcoma resection. There was positive correlation between serum VEGF and neutrophil counts, and negative between VEGF and hemoglobin content in animals with sarcoma. Animals with hemangiopericytoma showed higher serum VEGF levels compared to the patients with malignant peripheral nerve sheath. Circulating blood cells can contribute to elevate VEGF serum concentrations in dogs with soft tissue sarcomas and a possible role of VEGF in the angiogenesis of these tumors.     

Efficacy of Ceranock Attract and Kill System as a Control Method of Mediterranean Fruit Fly, Ceratitis capitata in Apricot Orchards in Central Iraq

A field experiment was conducted using Ceranock bait station, ＂attract and kill＂ system to combat Mediterranean fruit fly, Ceratitis capitata, in apricot orchards, Central of lraq during the 2013 field season. Control program was implemented in three orchards of mixed apricot varieties. Ceranock traps were hung in each tree of two separate orchards. Delta and McPhail traps were used to monitor Medfly population density in each treated and control orchards. Results indicated that Medfly activity in apricot orchards started during the last week of April coincided with the beginning of maturity for fruits of early varieties. The total of trapped insects in monitoring traps after one week of Ceranock application were 0, 1 and 31 adults for the 1st, 2nd and the control orchards, respectively. Three weeks later and on, the number of trapped adults started to increase dramatically. At the end of apricot season and after 45 days of Ceranock trap application, the number of trapped insects in monitoring traps reached 110, 111 and 2,349 adults/week, respectively, for the 1st, 2nd and 3rd orchard. The use of Ceranock traps reduced Medfly population density by 86%-97%. The percentage of fruit injuries in early maturing varieties were 0.75%, 1% and 34% for the 1st, 2nd and control orchards, respectively. While, late varieties were 2% and 3% for 1st and 2nd orchards, and 69% for the control. The percentage of reduction in fruit damages reached to 95% and 97%, respectively in the 1st and 2nd orchard. The results of this study demonstrate clearly the efficacy of Ceranock bait station, ＂attract and kill＂ system as a control measure for Medfly in apricot orchards.     

In vitro inhibitory effect of trichostatin A on canine grade 3 mast cell tumor

Mast cell tumor (MCT) is one of the most prevalent neoplasms that affect skin and soft tissue in dogs. Because mast cell tumors present a great variety of clinical appearance and behavior, their treatment becomes a challenge. Trichostatin A (TSA), an antifungal antibiotic, has shown inhibitory effects on the proliferation and induction of apoptosis in various types of cancer cells. In order to evaluate the potential of trichostatin A as a therapeutic drug, cells of grade 3 MCT were cultured and treated with concentrations of 1 nM to 400 nM of TSA. MTT assay and trypan blue exclusion assays were performed to estimate cell growth and cell viability, and cell cycle analysis was evaluated. TSA treatment showed a reduction in numbers of viable cells and an increase of cell death by apoptosis. The cell cycle analysis showed an increase of hypodiploid cells and a reduction of G0/G1 and G2/M –phases. According to these results, trichostatin A may be an interesting potential chemotherapeutic agent for the treatment of canine MCT.     

Clinical study of cryosurgery efficacy in the treatment of skin and subcutaneous tumors in dogs and cats

Objective— To evaluate the efficacy of cryosurgery for treatment of skin and subcutaneous tumors in dogs and cats.
Study Design— Prospective study.
Animals— Dogs (n=20), cats (10).
Methods— Cutaneous or subcutaneous tumors were treated by liquid nitrogen cryosurgical spray (1 cm from target tissue at 90° until a 5-mm halo of frozen tissue was achieved) for 15–60 seconds. Malignant lesions had 3 freeze–thaw cycles benign tumors, 2 cycles. The second or third freeze cycle was performed after complete thaw of the preceding freeze. Wounds healed by second intention. Follow-up was weekly for 1 month and then twice monthly until wounds healed, and final outcome was determined by telephone interview of owners.
Results— Tumor size ranged from 0.3 to 11 cm diameter with 28 (60%) being 0.3–1 cm; 8 (17%) 1.1–3 cm, and 11 (23%) >3.4 cm. Complications included edema, erythema and for extremity lesions, pain and lameness. Treated lesions (n=47) had an overall remission of 98% (mean follow-up, 345±172.02 days [range, 150–750 days]). One malignant peripheral nerve sheath tumor recurred 7 months after cryosurgical treatment.
Conclusion— Cryosurgery is an efficient method for treatment of skin and subcutaneous tumors in dogs and cats.
Clinical Relevance— Cryosurgical ablation is an effective means of treating small cutaneous or subcutaneous tumors in dogs and cats, especially in older animals where wound closure or cosmetic outcome might limit surgical excision alone.     

Brazilian biosafety law and the new breeding technologies

Globally, the area of land cultivated with genetically modified (GM) crops has increased a thousand-fold over the last two decades. Although this technology has become important for food production, the regulatory frameworks that underpin these outcomes are based on a list of requirements for a risk assessment that differ from country to country. In recent years, policymakers have had the opportunity to learn from the controversies over transgenics to create effective regulatory milestones for emerging technologies, allowing them to reach their potential for a more sustainable agriculture, ensuring food security. In Brazil, Law No. 11.105 of 24 March 2005 established a framework with four main organizations responsible for risk assessment and management. However, most of new breeding technologies did not exist at that time and were not considered in this law. In 2016, Normative Resolution No. 16 of the National Biosafety Technical Commission (CTNBio) was established to address this gap based on the evaluation of the products obtained through these techniques (termed Innovative Precision Improvement Techniques in the resolution), in a case-by-case consultation system. Briefly, if the product is designated to be a GM, the developer will have to go through the biosafety requirements and will be approved only after CTNBio risk assessment. If the product is designated not to be GM (for the purposes of the legislation), then it can be registered using the existing procedures. Currently, 152 GM products are commercially approved in Brazil. In 2018, CTNBio assessed the first consultation on commercial release of plants generated using the new breeding technologies and has subsequently approved six products. It is expected that many institutions would be able to participate in Brazilian and world markets, developing and introducing new biotechnological solutions and products through a more sustainable approach and without facing public disapproval, a common issue for GM crops.     

Photo-Assisted Degradation,Toxicological Assessment,and Modeling Using Artificial Neural Networks of Reactive Gray BF-2R Dye

This work investigates the degradation of Reactive Gray BF-2R dye (a blend of reactive yellow 145, reactive orange 122 and reactive black 5 dyes) using UV/H₂O₂, Fenton, and photo-Fenton-advanced oxidative processes, with artificial sunlight and UV-C radiations. The photo-Fenton process employing UV-C radiation was the most efficient under the conditions studied. The ideal conditions for the degradation of the dye, determined using a factorial design 2³ and a study of the concentration of hydrogen peroxide ([H₂O₂]), were [H₂O₂] equal to 40 mg L^?1, iron concentration [Fe] of 1 mg L^?1, and pH between 3 and 4. The Chan and Chu non-linear kinetic model predicted the kinetic data with a degradation of over 98% for color and 68% for aromatics after 60 min. The behavior of the chemical oxygen demand fitted the first-order kinetic model well, with a degradation of 64% after 60 min. The Multilayer Perceptron 7-11-2 artificial neural network model enabled to model the degradation process of the aromatics and accurately predict the experimental data. Toxicity tests indicated that the post-treatment samples were non-toxic for Escherichia coli bacteria, and Portulaca grandiflora and Basil sabory seeds. However, they inhibited the growth of Lactuca sativa seeds and Salmonella enteritidis bacteria. The photo-Fenton process with UV-C radiation degraded the dye studied efficiently and the degradation percentages were, on average, 7% and 5% higher for color than those observed when employing the Fenton and UV/H₂O₂ processes, respectively. With the aromatic, however, they were 84% and 62% higher, thus justifying the use of this process.     

Influence of potassium nutrition and the nitrate/ammonium ratio on the putrescine and spermidine contents in banana vitroplants

Potassium (K) is required in high doses by the banana (Musa sp.) plant and interacts with other nutrient elements in which banana tissues are maintained under in vitro condition as a consequence modifications in the plant metabolism take place mainly in nitrogen (N) compounds, such as proteins, amino acids, and secondary compounds. When K is present in concentrations lower than that required, diamines such as putrescine and poliamines are formed. This metabolic disorder can also be correlated with the presence of different inorganic N forms, such as nitrate (NO₃) and ammonium (NH₄), and the ratios between both ions as well. In order to follow the physiological performance of the interrelationships, K/putrescine and of the NO₃/NH₄ ratio in the tissue of banana vitroplantlets, shoot apex of two banana cvs. Nanica and Prata Ana were maintained in modified MS medium in the presence of six different doses of K: 5, 10, 15, 20, 25, and 30 mM. After the period of tissue proliferation the cultures were transferred to rooting media containing the same different K doses. Dry matter, K, putrescine, and spermidine contents and their accumulation were determined in the shoots and roots of the vitroplantlets and in the shoot apex of the expiant donor cultivar as well as the corresponding values for the whole vitroplantlets calculated. The data were statistically analyzed. The contents and accumulations of putrescine and spermidine in banana tissues were enhanced as K concentration decreased in the medium: four times (0.19% of the dry matter) for cv. Nanica and eight times (0.25% of the dry matter) for cv. Prata Ana. This behavior was not only related to the K depletion but to the NO₃/NH₄ ratio as well.     

Cell proliferation and expression of connexins differ in melanotic and amelanotic canine oral melanomas

Melanoma is a malignant neoplasm occurring in several animal species, and is the most frequently found tumor in the oral cavity in dogs. Melanomas are classified into two types: melanotic and amelanotic. Prior research suggests that human amelanotic melanomas are more aggressive than their melanotic counterparts. This study evaluates the behavior of canine melanotic and amelanotic oral cavity melanomas and quantifies cell proliferation and the expression of connexins. Twenty-five melanomas (16 melanotic and 9 amelanotic) were collected from dogs during clinical procedures at the Veterinary Hospital of the School of Veterinary Medicine and Animal Science of the University of São Paulo, Brazil. After diagnosis, dogs were followed until death or euthanasia. Histopathology confirmed the gross melanotic or amelanotic characteristics and tumors were classified according to the WHO. HMB45 or Melan A immunostainings were performed to confirm the diagnosis of amelanotic melanomas. Cell proliferation was quantified both by counting mitotic figures and PCNA positive nuclei. Expressions of connexins 26 and 43 were evaluated by immunohistochemistry, qRT-PCR and Western blot. Dogs bearing amelanotic melanomas presented a shorter lifespan in comparison to those with melanotic melanomas. Cell proliferation was significantly higher in amelanotic melanomas. Expressions of Connexins 26 and 43 were significantly reduced in amelanotic melanomas. The results presented here suggest that oral cavity melanotic and amelanotic melanomas differ regarding their behavior, cell proliferation and connexin expression in dogs, indicating a higher aggressiveness of amelanotic variants.